Novel bdellovibrio bacteriovorus having antibiotic adaptability, and use thereof

Abstract

The present invention relates to a Bdellovibrio bacteriovorus 109J (Km240T) strain with accession No. KCTC15671BP, having antibiotic adaptability, and, more specifically, to a Bdellovibrio bacteriovorus 109J (Km240T) strain with accession No. KCTC15671BP, having antibiotic adaptability, the Bdellovibrio bacteriovorus 109J (Km240T) strain with accession No. KCTC15671BP being capable of effectively eliminating pathogens and removing gDNA and antibiotic resistance genes of pathogens, thereby preventing the spread of the pathogens, and thus present invention is expected to have synergistic effects when used with antibiotics.

Description

Novel delobibrio bacteriovorus having antibiotic adaptability and uses thereof

[0001] The present invention relates to a novel Dellovibrio bacteriovorus having antibiotic adaptability and its use.

[0002]

[0003] Conventional antibiotic-based methods for eliminating antibiotic-resistant pathogens (superbugs) are limited by their inherent resistance to antibiotics, making them ineffective. Furthermore, excessive antibiotic use can promote the emergence of antibiotic-resistant pathogens.

[0004] Accordingly, there is a need for technologies that can reduce antibiotic usage and prevent the spread of antibiotic resistance genes in antibiotic-resistant pathogens. The present inventors propose a novel strain of Dellovibrio bacteriovorus 109J (Km240T) with antibiotic adaptability, as well as a method for producing and using the same.

[0005]

[0006] The present invention aims to provide a novel Dellovibrio bacteriovorus 109J (Km240T) having antibiotic adaptability.

[0007] The present invention aims to provide an antibiotic composition comprising a novel Dellovibrio bacteriovorus 109J (Km240T) having antibiotic adaptability.

[0008] The present invention aims to provide a method for producing a novel Dellovibrio bacteriovorus 109J (Km240T) having antibiotic adaptability and a method for killing pathogens using the same.

[0009]

[0010] 1. Dellovibrio bacteriovorus 109J (Km240T) strain with accession number KCTC15671BP, ​​which has antibiotic adaptability.

[0011] 2. In the above 1, the antibiotic is a strain selected from the group consisting of kanamycin, gentamicin, ciprofloxacin or a pharmaceutically acceptable salt thereof.

[0012] 3. An antibiotic composition for a pathogen comprising a strain of Dellovibrio bacteriovorus 109J (Km240T) having antibiotic adaptability and having the deposit number KCTC15671BP.

[0013] 4. An antibiotic composition for a pathogen, further comprising an antibiotic selected from the group consisting of kanamycin, gentamicin, ciprofloxacin or a pharmaceutically acceptable salt thereof, in the above 3.

[0014] 5. In the above 3, an antibiotic composition for a pathogen selected from the group consisting of Acinetobacter baumannii, Escherichia coli, and Klebsiella pneumoniae.

[0015] 6. A method for killing a pathogen, comprising a step of treating an antibiotic-resistant pathogen with a strain of Dellovibrio bacteriovorus 109J (Km240T) having the deposit number KCTC15671BP.

[0016] 7. A method for killing a pathogen, comprising a step of further treating with an antibiotic selected from the group consisting of kanamycin, gentamicin, ciprofloxacin or a pharmaceutically acceptable salt thereof, in the above 6.

[0017] 8. In the above 6, the method for killing a pathogen is to treat a cell or an animal other than a human.

[0018] 9. A method for producing a strain of Dellovibrio bacteriovorus 109J (Km240T), comprising: a first step of culturing Dellovibrio bacteriovorus in a first medium treated with a pathogen and antibiotics; a second step of confirming whether the absorbance of the first medium is less than 0.5; and a third step of isolating Dellovibrio bacteriovorus from the first medium when the absorbance of the second step is less than 0.5 and recultivating it in a second medium having twice the concentration of antibiotics.

[0019] 10. In the above 9, the initial absorbance of the first medium is 1, a method for producing a strain of Dellovibrio bacteriovorus 109J (Km240T).

[0020] 11. A method for producing a strain of Dellovibrio bacteriovorus 109J (Km240T) in the above 9, wherein the absorbance is measured at 600 nm.

[0021] 12. In the above 9, the pathogen is a method for producing a strain of Dellovibrio bacteriovorus 109J (Km240T) selected from the group consisting of Acinetobacter baumannii, Escherichia coli, and Klebsiella pneumoniae.

[0022] 13. A method for producing a strain of Dellovibrio bacteriovorus 109J (Km240T), wherein the antibiotic in the above 9 is selected from the group consisting of kanamycin, gentamicin, ciprofloxacin or a pharmaceutically acceptable salt thereof.

[0023]

[0024] The present invention provides a novel Dellovibrio bacteriovorus 109J (Km240T) having antibiotic adaptability, which can effectively eliminate pathogens.

[0025] The present invention can completely eliminate pathogens by using a novel Dellovibrio bacteriovorus 109J (Km240T) having antibiotic adaptability together with antibiotics.

[0026] The present invention can prevent the spread of pathogens by removing their gDNA and antibiotic resistance genes.

[0027] The method of the present invention can produce a novel Dellovibrio bacteriovorus 109J (Km240T) having high antibiotic adaptability and whose predation and removal ability is not reduced even when used together with antibiotics.

[0028] The present invention can be effectively utilized for the elimination of antibiotic-resistant pathogens.

[0029] The method of the present invention can be effectively utilized in the manufacture of an antibiotic composition for eliminating antibiotic-resistant pathogens.

[0030] The present invention can contribute to preventing the emergence of antibiotic-resistant pathogens by reducing the amount of antibiotics used.

[0031]

[0032] Figure 1 shows the results of antibiotic adaptation induced by long-term, continuous single exposure of Dellovibrio bacteriovorus 109J wild type to various concentrations of the antibiotic kanamycin. The Y-axis is OD (Optical Density, 600 nm), and the decrease in OD of the pathogen is an indicator of growth and proliferation of Dellovibrio bacteriovorus 109J (Km240T).

[0033] Figure 2 shows the results of gradually exposing wild type Dellovibrio bacteriovorus 109J to antibiotics to induce antibiotic adaptation. When the OD of E. coli decreased to less than 0.5, it indicates that Dellovibrio bacteriovorus 109J grew. The circled portion represents the Dellovibrio bacteriovorus 109J that grew in 24-hour units, and the selected samples were re-cultured in a diluted E. coli solution containing 0 to 240 μg / ml of kanamycin, and the OD was measured the next day. The circle in the upper right corner means that all samples adapted to 240 μg / ml of kanamycin in 7 days.

[0034] Figure 3 shows the results of measuring the OD of E. coli (K12 MG1655) treated with each sample according to the concentration (㎍ / ㎖) of kanamycin used in the predation test. CTRL is E. coli only, WT is wild type Dellovibrio bacteriovorus 109J, KmR is kanamycin-adapted Dellovibrio bacteriovorus 109J (Km240T), KmR Revert is kanamycin-adapted Dellovibrio bacteriovorus 109J (Km240T) cultured for 14 days under antibiotic-free conditions, pSUP404.2 is Dellovibrio bacteriovorus 109J carrying the kanamycin-adapted plasmid pSUP404.2, and pSUP404.2 Revert is Dellovibrio bacteriovorus 109J carrying the kanamycin-adapted plasmid pSUP404.2 cultured for 14 days under antibiotic-free conditions.

[0035] Figure 4 shows the results of measuring the viability of pathogens remaining 24 hours after administering kanamycin alone or kanamycin and kanamycin-adapted Dellovibrio bacteriovorus 109J (Km240T) together to E. coli NCCP 16044, a pathogen sensitive to kanamycin.

[0036] Figure 5 shows the results of measuring the viability of the remaining pathogens 24 hours after administering kanamycin alone or co-administering kanamycin and kanamycin-adapted Dellovibrio bacteriovorus 109J (Km240T) to K. pneumoniae YS-CRE-16201, an intermediate-resistant pathogen to kanamycin.

[0037] Figure 6 shows the results of measuring the viability of pathogens remaining 24 hours after administering kanamycin alone or kanamycin and kanamycin-adapted Dellovibrio bacteriovorus 109J (Km240T) to Acinetobacter baumannii (A. baumanniiNCCP 15994), a kanamycin-resistant pathogen.

[0038] Figure 7 shows the results of measuring the amount of DNA (16S rDNA in gDNA or carbapenem antibiotic resistance gene KPC-2) of Escherichia coli (E. coliNCCP 16044), a carbapenem-resistant pathogen, when only kanamycin was used (left) and when kanamycin and Dellovibrio bacteriovorus 109J (Km240T) were used together.

[0039] Figure 8 shows the results of measuring the amount of DNA (gltA or carbapenem antibiotic resistance gene KPC-2 in gDNA) of K. pneumoniae YS-CRE-16201, a carbapenem-resistant pathogen, in each case of using only kanamycin (left) and using kanamycin and Dellovibrio bacteriovorus 109J (Km240T) together.

[0040] Figure 9 shows the results of measuring the amount of DNA (16s rDNA in gDNA or carbapenem antibiotic resistance gene Oxa-23) of Acinetobacter baumannii (A. baumanniiNCCP 15994), a carbapenem-resistant pathogen, when only kanamycin was used (left) and when kanamycin and Dellovibrio bacteriovorus 109J (Km240T) were used together.

[0041]

[0042] The present invention provides a strain of Dellovibrio bacteriovorus 109J (Km240T) having antibiotic adaptability and having the deposit number KCTC15671BP.

[0043] Antibiotics are not limited to a specific antibiotic.

[0044] The antibiotic may be any one selected from the group consisting of kanamycin, gentamicin, ciprofloxacin or a pharmaceutically acceptable salt thereof.

[0045] The present invention provides an antibiotic composition capable of effectively eliminating pathogens by including the strain Dellovibrio bacteriovorus 109J (Km240T) having the deposit number KCTC15671BP.

[0046] The present invention provides an antibiotic composition for a pathogen, further comprising an antibiotic selected from the group consisting of kanamycin, gentamicin, ciprofloxacin, or a pharmaceutically acceptable salt thereof, in addition to the Dellovibrio bacteriovorus 109J (Km240T) strain having the deposit number KCTC15671BP.

[0047] Pathogens are not limited to specific pathogens.

[0048] In one embodiment, the pathogen may be any one selected from the group consisting of Acinetobacter baumannii, Escherichia coli, and Klebsiella pneumoniae.

[0049] The present invention provides a method for killing a pathogen, comprising the step of treating an antibiotic-resistant pathogen with a strain of Dellovibrio bacteriovorus 109J (Km240T) having the deposit number KCTC15671BP.

[0050] The present invention provides a method for killing a pathogen, comprising the step of treating an antibiotic-resistant pathogen with a strain of Dellovibrio bacteriovorus 109J (Km240T) having the deposit number KCTC15671BP, ​​and then further treating the pathogen with any one antibiotic selected from the group consisting of kanamycin, gentamicin, ciprofloxacin, or a pharmaceutically acceptable salt thereof.

[0051] The present invention provides a strain of Dellovibrio bacteriovorus 109J (Km240T) with the accession number KCTC15671BP, ​​which can prevent the spread of pathogens by removing gDNA and antibiotic resistance genes, and has antibiotic adaptability that is expected to have a synergistic effect when used together with antibiotics.

[0052] The present invention provides a method for producing a strain of Dellovibrio bacteriovorus 109J (Km240T) having the deposit number KCTC15671BP.

[0053] The method for producing the Dellovibrio bacteriovorus 109J (Km240T) strain of the present invention comprises a first step of culturing Dellovibrio bacteriovorus in a first medium treated with a pathogen and an antibiotic; a second step of confirming whether the absorbance of the first medium is less than 0.5; and a third step of isolating Dellovibrio bacteriovorus from the first medium when the absorbance of the second step is less than 0.5 and re-cultivating it in a second medium having twice the concentration of the antibiotic.

[0054] Step 1 involves culturing antibiotic-susceptible Dellovibrio bacteriovorus on antibiotic-treated media. The media contains both the antibiotic and the pathogens that serve as food for Dellovibrio bacteriovorus.

[0055] Antibiotics need not be treated at a specific concentration, as long as they are treated at a concentration that allows Dellovibrio bacteriovorus to exhibit antibiotic adaptability in the medium. In one embodiment, the antibiotics may be treated at a concentration of 1 to 500 μg / ml.

[0056] Pathogens are not limited to a specific type of pathogen, as long as they are food for Dellovibrio bacteriovorus. Pathogens include bacteria, viruses, fungi, prions, other parasites, and algae. In one embodiment, the pathogen is Escherichia coli.

[0057] Step 2 is to determine whether Dellovibrio bacteriovorus survived the culture in Step 1. If Dellovibrio bacteriovorus survives by consuming pathogens despite antibiotic treatment, the amount of pathogens will decrease, resulting in a decrease in the absorbance of the first medium.

[0058] The initial absorbance of the first medium is 1, and as the pathogen decreases, the absorbance drops to, for example, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, etc. When measuring the absorbance at 600 nm, antibiotics (compounds) do not affect the absorbance, and Dellovibrio bacteriovorus has some effect, but it is negligible in this experiment, and the amount of pathogens mainly affects the absorbance.

[0059] If the absorbance of the first medium falls below 0.5, this indicates that approximately 50% of the initial pathogens have been killed, and Dellovibrio bacteriovorus have acquired antibiotic adaptability and multiplied. In the second step, the Dellovibrio bacteriovorus that have acquired antibiotic adaptability are selected and moved on to the third step.

[0060] The third stage is to isolate the Dellovibrio bacteriovorus selected in the second stage and re-culture them in a second medium treated with antibiotics at a concentration twice as high as that of the first medium. Because the antibiotic concentration is high in the third stage, only the Dellovibrio bacteriovorus strains with relatively high antibiotic adaptability or those that have adapted to the twice-higher concentration of antibiotics in the second stage and acquired improved antibiotic adaptability survive.

[0061] After step 3, a step of measuring the absorbance can be performed. By confirming that the concentration (absorbance) of the pathogen is less than half despite treatment with a higher concentration of antibiotic, Dellovibrio bacteriovorus that have acquired antibiotic adaptability can be selected again.

[0062] In one embodiment, the method for producing the Dellovibrio bacteriovorus 109J (Km240T) strain of the present invention may include the steps of culturing Dellovibrio bacteriovorus 109J after treating a medium with a pathogen (such as Escherichia coli) and 3.75 ㎍ / ㎖ of an antibiotic (kanamycin); selecting a Dellovibrio bacteriovorus 109J cultured sample having an Optical Density value of the pathogen measured at 600 nm of less than 0.5 after the culturing; and culturing Dellovibrio bacteriovorus 109J by treating the selected sample with an antibiotic concentration twice as high.

[0063] Hereinafter, the present invention will be described in more detail with examples.

[0064]

[0065] Example

[0066] Example 1: Method 1 for producing Dellovibrio bacteriovorus 109J (Km240T)

[0067] Dellovibrio bacteriovorus 109J was exposed to various concentrations of the antibiotic kanamycin for a long period of time to induce antibiotic adaptation. The manufacturing method for this is as follows. First, a colony of E. coli (K12 MG1655) cultured on an LB agar plate was inoculated into LB medium and cultured overnight. Then, only E. coli cells were separated from the culture medium using a centrifuge (4,000 x g, 10 min), and then diluted with sterilized 25 mM HEPES buffer containing 3 mM MgCl2, 2 mM CaCl2, and 0.8 g / L Nutrient broth, and the optical density value was adjusted to 1.1. OD was measured using an Eppendorf Biophotometer, and the value of CFU / ㎖ at this time was approximately 5 X 10 8It was. Dellovibrio bacteriovorus 109J cultured in advance in this dilution was passed through a 0.45㎛ PES filter, and 1.5㎖ of it was administered to 13.5㎖ of the corresponding feed dilution, and then kanamycin was administered to the target concentrations (0, 30, 60, 120, 240㎍ / ㎖). The corresponding culture was continuously cultured in a 30℃ shaking incubator (300 rpm), and the OD was measured at 24-hour intervals. As a result, in the case of low concentration of kanamycin (30㎍ / ㎖), the antibiotic-adapted Dellovibrio bacteriovorus 109J (Km240T) grew to saturation on the 3rd to 4th day, which was confirmed by the decrease in the Optical Density (600㎚) value of Escherichia coli (E. coliK12 MG1655), the pathogen used in the experiment. In the case of high concentration of kanamycin (240 μg / ml), 1 of the triplicate samples of Dellovibrio bacteriovorus 109J (Km240T) was confirmed to have grown to saturation on the 7th day (Fig. 1).

[0068]

[0069] Example 2: Method 2 for producing Dellovibrio bacteriovorus 109J (Km240T)

[0070] Dellovibrio bacteriovorus 109J was gradually exposed to antibiotics to induce antibiotic adaptation. The concentrations of kanamycin used at this time were 0, 3.75, 7.5, 15, 30, 60, 120, and 240 ㎍ / ㎖. The method for preparing E. coli and Dellovibrio bacteriovorus 109J for this example is the same as Example 1. The decrease in the Optical Density (600 nm) value of E. coli (E. coliK12 MG1655), the pathogen used in the experiment, was measured every 24 hours, and a sample with an antibiotic concentration at which the OD600 value fell below 0.5 (an antibiotic concentration at which the growth of Dellovibrio bacteriovorus 109J was inhibited by less than 50% (IC50; indicated by a circle)) was selected. Selected samples were re-cultured using the method of Example 1 in the presence of fresh pathogens and fresh kanamycin (0, 3.75, 7.5, 15, 30, 60, 120, 240 μg / ml) to observe increased antibiotic adaptability. It was confirmed that all triplicates of Dellovibrio bacteriovorus 109J had adapted to the 240 μg / ml concentration of kanamycin after 7 days.

[0071] These experimental results showed that it was possible to more stably provide high adaptability to antibiotics to Dellovibrio bacteriovorus 109J.

[0072]

[0073] Example 3: OD measurement experiment for each sample according to kanamycin concentration

[0074] Six types of samples were treated with the pathogenic bacterium Escherichia coli (E. coli K12 MG1655), and the OD values ​​of the E. coli according to the predation time were measured after treating each sample with different concentrations (㎍ / ㎖) of kanamycin. The method for preparing the E. coli and the predation bacteria for this purpose was used with some modifications of the method used in Example 1. Since this experiment was conducted using a transparent 96-well plate instead of a test tube, the final volume of the test solution was reduced to 200 ㎕. In addition, the OD 600 nm value of the E. coli was measured using a microplate reader (TECAN spark, Switzerland).

[0075] When antibiotic adaptability was granted, the predation ability was slightly inhibited compared to the wild type (WT) (compare WT and KmR in the first graph of Fig. 3), but it was confirmed that the predation ability recovered to the same level as the wild type (WT) by culturing under antibiotic-free conditions for 14 days (compare WT and KmR Revert in the first graph of Fig. 3).

[0076] In the case of Dellovibrio bacteriovorus 109J (Km240T) that was endowed with antibiotic adaptability, it was confirmed that it did not lose its predatory ability under antibiotic conditions even after being cultured under antibiotic-free conditions for 14 days, but rather had an even greater predatory ability (compare KmR and KmR Revert in the second and third graphs of Fig. 3).

[0077] However, it was confirmed that Dellovibrio bacteriovorus 109J, which was endowed with kanamycin adaptability through a plasmid (pSUP404.2), lost some of its adaptability when cultured under antibiotic-free conditions for 14 days (compare pSUP404.2 Revert and pSUP404.2 in the second and third graphs of Fig. 3).

[0078]

[0079] Example 4: Comparative experiment on the pathogen killing effect of kanamycin alone and co-administration of kanamycin and Dellovibrio bacteriovorus 109J (Km240T)

[0080] (1) If the pathogen is E. coli (E. coliNCCP 16044) that is sensitive to kanamycin

[0081] Escherichia coli (E. coliNCCP 16044), a pathogen sensitive to kanamycin, was administered kanamycin alone or together with kanamycin and kanamycin-adapted Dellovibrio bacteriovorus 109J (Km240T), and the viability of the remaining pathogens after 24 hours was measured. The experimental method for this was the same as Example 1, and viability was measured by serially diluting the culture solution in HEPES buffer, spreading it on an LB agar plate, counting the number of colonies after 24 hours, and calculating backward by the dilution factor.

[0082] It was confirmed that a smaller amount of pathogens remained when kanamycin and kanamycin-adapted Dellovibrio bacteriovorus 109J (Km240T) were co-administered compared to when kanamycin was administered alone (Fig. 4).

[0083] (2) If the pathogen is K. pneumoniae YS-CRE-16201, which is intermediately resistant to kanamycin.

[0084] K. pneumoniae YS-CRE-16201, an intermediate-resistant pathogen to kanamycin, was administered with kanamycin alone or together with kanamycin and kanamycin-adapted Dellovibrio bacteriovorus 109J (Km240T), and the viability of the remaining pathogens after 24 hours was measured. The experimental method for this was the same as Example 1, and viability was measured by serially diluting the culture solution in HEPES buffer, spreading it on an LB agar plate, counting the number of colonies after 24 hours, and calculating backward by the dilution factor.

[0085] Compared to when kanamycin was administered alone, co-administration of kanamycin and kanamycin-adapted Dellovibrio bacteriovorus 109J (Km240T) resulted in fewer pathogens remaining (Fig. 5). In particular, co-administration demonstrated a synergistic effect at kanamycin concentrations of 60 μg / ml or higher, resulting in complete elimination of the pathogens.

[0086] (3) If the pathogen is Acinetobacter baumannii (A. baumanniiNCCP 15994) that is resistant to kanamycin.

[0087] Kanamycin alone or in combination with kanamycin-adapted Dellovibrio bacteriovorus 109J (Km240T) was administered to Acinetobacter baumannii (A. baumanniiNCCP 15994), a kanamycin-resistant pathogen, and the viability of the remaining pathogens after 24 hours was measured. The experimental method for this was the same as Example 1, and viability was measured by serially diluting the culture solution in HEPES buffer, spreading it on an LB agar plate, counting the number of colonies after 24 hours, and calculating backward by the dilution factor.

[0088] Compared to when kanamycin was administered alone, when kanamycin and kanamycin-adapted Dellovibrio bacteriovorus 109J (Km240T) were co-administered, fewer pathogens remained (Fig. 6). Notably, although kanamycin did not kill any pathogens, it was confirmed that approximately 99.9% of the pathogens were killed by the predatory ability of the predatory bacteria.

[0089]

[0090] Example 5: Comparative experiment on the pathogen DNA removal effect of kanamycin alone and co-administration of kanamycin and Dellovibrio bacteriovorus 109J (Km240T)

[0091] (1) In the case of E. coli (E. coliNCCP 16044), a carbapenem-resistant pathogen

[0092] After administering kanamycin alone or co-administering kanamycin and kanamycin-adapted Dellovibrio bacteriovorus 109J (Km240T) to Escherichia coli, the amount of DNA (16S rDNA in gDNA or carbapenem antibiotic resistance gene KPC-2) in the E. coli was measured.

[0093] When kanamycin-adapted Dellovibrio bacteriovorus 109J (Km240T) was administered (Predated, right) compared to the untreated control group (Unpredated, Kanamycin 0 ㎍ / ㎖), it was confirmed through qPCR that gDNA or antibiotic resistance genes of E. coli were removed by approximately 99% (Fig. 7). 1 ㎕ of bacterial culture was pretreated at 95°C for 10 minutes and then used for qPCR analysis. The qPCR machine used was LightCycler® 480 (Roche, Switzerland), and the mastermix used was SYBR Green.

[0094] (2) In the case of Klebsiella pneumoniae (K. pnuemoniaeYS-CRE-16201), a carbapenem-resistant pathogen

[0095] After administering kanamycin alone or in combination with kanamycin and kanamycin-adapted Dellovibrio bacteriovorus 109J (Km240T) to K. pneumoniae, the amount of DNA (gltA or carbapenem antibiotic resistance gene KPC-2 in gDNA) of K. pneumoniae was measured.

[0096] Compared to the untreated control group (Unpredated, Kanamycin 0 ㎍ / ㎖), when administered with kanamycin-adapted Dellovibrio bacteriovorus 109J (Km240T) (Predated, right), approximately 99% of the gDNA or antibiotic resistance genes of Klebsiella pneumoniae were removed, as confirmed by qPCR (Fig. 8). 1 ㎕ of bacterial culture was pretreated at 95℃ for 10 min and used for qPCR analysis. The qPCR machine was LightCycler® 480 (Roche, Switzerland), and the mastermix was SYBR Green.

[0097] (3) In the case of Acinetobacter baumannii (A. baumanniiNCCP 15994), a carbapenem-resistant pathogen

[0098] Acinetobacter baumannii was administered kanamycin alone or in combination with kanamycin and kanamycin-adapted Dellovibrio bacteriovorus 109J (Km240T), and the amount of DNA (16s rDNA in gDNA or carbapenem antibiotic resistance gene Oxa-23) in Acinetobacter baumannii was measured.

[0099] Compared to the untreated control group (Unpredated, Kanamycin 0 ㎍ / ㎖), when administered kanamycin-adapted Dellovibrio bacteriovorus 109J (Km240T) (Predated, right), it was confirmed through qPCR that the gDNA or antibiotic resistance gene of Acinetobacter baumannii was removed by approximately 99% (Fig. 9). 1 ㎕ of bacterial culture was pretreated at 95℃ for 10 min and used for qPCR analysis. The qPCR machine was LightCycler® 480 (Roche, Switzerland), and the mastermix was SYBR Green.

[0100]

[0101] Example 6: Comparative experiment on the adaptability of Dellovibrio bacteriovorus 109J wild type and Dellovibrio bacteriovorus 109J (Km240T) to antibiotics

[0102] The adaptability of wild-type Dellovibrio bacteriovorus 109J and Dellovibrio bacteriovorus 109J (Km240T) strains treated with different antibiotics was compared. The experimental preparation for this was the same as that of Example 3. The results are shown in Table 1 below.

[0103] Antibiotics: Delovibriobacteriovorus 109J wild type (㎍ / ㎖) Delovibriobacteriovorus 109J (Km240T) (㎍ / ㎖) Kanamycin 4240 or more (60 times or more) Gentamicin 0.632.5 (4 times) Ciprofloxacin 0.51 (2 times)

[0104]

[0105] Below, the deposit certificate and translation of the deposit certificate for Dellovibrio bacteriovorus 109J (Km240T) are attached.

[0106]

Claims

1. A strain of Dellovibrio bacteriovorus 109J (Km240T) with the accession number KCTC15671BP having antibiotic adaptability.

2. In claim 1, the antibiotic is a strain selected from the group consisting of kanamycin, gentamicin, ciprofloxacin or a pharmaceutically acceptable salt thereof.

3. An antibiotic composition for a pathogen, comprising a strain of Dellovibrio bacteriovorus 109J (Km240T) having antibiotic adaptability and having the deposit number KCTC15671BP.

4. An antibiotic composition for a pathogen, further comprising an antibiotic selected from the group consisting of kanamycin, gentamicin, ciprofloxacin or pharmaceutically acceptable salts thereof, in claim 3.

5. An antibiotic composition for a pathogen according to claim 3, wherein the pathogen is any one of Acinetobacter baumannii, Escherichia coli, and Klebsiella pneumoniae.

6. A method for killing a pathogen, comprising the step of treating an antibiotic-resistant pathogen with a strain of Dellovibrio bacteriovorus 109J (Km240T) having the deposit number KCTC15671BP.

7. A method for killing a pathogen, comprising the step of further treating with an antibiotic selected from the group consisting of kanamycin, gentamicin, ciprofloxacin or a pharmaceutically acceptable salt thereof, in claim 6.

8. A method for killing a pathogen according to claim 6, wherein the strain is applied to a cell or an animal other than a human.

9. The first step is to cultivate Dellovibrio bacteriovorus in the first medium treated with pathogens and antibiotics; A second step of checking whether the absorbance of the first badge is less than 0.5; and A method for producing a Dellovibrio bacteriovorus 109J (Km240T) strain, comprising a third step of isolating the Dellovibrio bacteriovorus from the first medium when the absorbance of the second step is less than 0.5 and re-cultivating it in a second medium having twice the concentration of the antibiotic.

10. A method for producing a Dellovibrio bacteriovorus 109J (Km240T) strain according to claim 9, wherein the initial absorbance of the first medium is 1.

11. A method for producing a Dellovibrio bacteriovorus 109J (Km240T) strain according to claim 9, wherein the absorbance is measured at 600 nm.

12. A method for producing a Dellovibrio bacteriovorus 109J (Km240T) strain according to claim 9, wherein the pathogen is any one selected from the group consisting of Acinetobacter baumannii, Escherichia coli, and Klebsiella pneumoniae.

13. A method for producing a Dellovibrio bacteriovorus 109J (Km240T) strain, wherein the antibiotic according to claim 9 is any one selected from the group consisting of kanamycin, gentamicin, ciprofloxacin or pharmaceutically acceptable salts thereof.

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