CN114395610A - Method for rapidly screening bacteriocin-producing microorganisms - Google Patents
Method for rapidly screening bacteriocin-producing microorganisms Download PDFInfo
- Publication number
- CN114395610A CN114395610A CN202210068639.8A CN202210068639A CN114395610A CN 114395610 A CN114395610 A CN 114395610A CN 202210068639 A CN202210068639 A CN 202210068639A CN 114395610 A CN114395610 A CN 114395610A
- Authority
- CN
- China
- Prior art keywords
- sample
- bacteriocin
- producing
- screening
- flat plate
- Prior art date
- Legal status (The legal status 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 status listed.)
- Pending
Links
- 238000012216 screening Methods 0.000 title claims abstract description 48
- 108010062877 Bacteriocins Proteins 0.000 title claims abstract description 40
- 244000005700 microbiome Species 0.000 title claims abstract description 32
- 238000000034 method Methods 0.000 title claims abstract description 23
- 241000894006 Bacteria Species 0.000 claims abstract description 26
- 229920001817 Agar Polymers 0.000 claims abstract description 23
- 239000008272 agar Substances 0.000 claims abstract description 23
- 238000012258 culturing Methods 0.000 claims abstract description 16
- 239000011248 coating agent Substances 0.000 claims abstract description 14
- 238000000576 coating method Methods 0.000 claims abstract description 14
- 239000002504 physiological saline solution Substances 0.000 claims abstract description 14
- 239000006916 nutrient agar Substances 0.000 claims abstract description 13
- 239000000126 substance Substances 0.000 claims abstract description 12
- 239000000203 mixture Substances 0.000 claims abstract description 7
- 239000012895 dilution Substances 0.000 claims abstract description 3
- 238000010790 dilution Methods 0.000 claims abstract description 3
- 241000193755 Bacillus cereus Species 0.000 claims description 13
- 241000186779 Listeria monocytogenes Species 0.000 claims description 13
- 241000191967 Staphylococcus aureus Species 0.000 claims description 13
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 3
- 239000012898 sample dilution Substances 0.000 claims description 2
- 230000005764 inhibitory process Effects 0.000 abstract description 13
- 244000000010 microbial pathogen Species 0.000 abstract description 7
- 230000000844 anti-bacterial effect Effects 0.000 abstract description 4
- 239000000523 sample Substances 0.000 description 30
- 230000000694 effects Effects 0.000 description 12
- 239000012470 diluted sample Substances 0.000 description 10
- 230000002401 inhibitory effect Effects 0.000 description 9
- 230000003385 bacteriostatic effect Effects 0.000 description 6
- 108010053775 Nisin Proteins 0.000 description 5
- NVNLLIYOARQCIX-MSHCCFNRSA-N Nisin Chemical compound N1C(=O)[C@@H](CC(C)C)NC(=O)C(=C)NC(=O)[C@@H]([C@H](C)CC)NC(=O)[C@@H](NC(=O)C(=C/C)/NC(=O)[C@H](N)[C@H](C)CC)CSC[C@@H]1C(=O)N[C@@H]1C(=O)N2CCC[C@@H]2C(=O)NCC(=O)N[C@@H](C(=O)N[C@H](CCCCN)C(=O)N[C@@H]2C(NCC(=O)N[C@H](C)C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCSC)C(=O)NCC(=O)N[C@H](CS[C@@H]2C)C(=O)N[C@H](CC(N)=O)C(=O)N[C@H](CCSC)C(=O)N[C@H](CCCCN)C(=O)N[C@@H]2C(N[C@H](C)C(=O)N[C@@H]3C(=O)N[C@@H](C(N[C@H](CC=4NC=NC=4)C(=O)N[C@H](CS[C@@H]3C)C(=O)N[C@H](CO)C(=O)N[C@H]([C@H](C)CC)C(=O)N[C@H](CC=3NC=NC=3)C(=O)N[C@H](C(C)C)C(=O)NC(=C)C(=O)N[C@H](CCCCN)C(O)=O)=O)CS[C@@H]2C)=O)=O)CS[C@@H]1C NVNLLIYOARQCIX-MSHCCFNRSA-N 0.000 description 5
- 239000004309 nisin Substances 0.000 description 5
- 235000010297 nisin Nutrition 0.000 description 5
- 238000009792 diffusion process Methods 0.000 description 3
- 239000000022 bacteriostatic agent Substances 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 238000004166 bioassay Methods 0.000 description 1
- 230000029918 bioluminescence Effects 0.000 description 1
- 238000005415 bioluminescence Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000013537 high throughput screening Methods 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- General Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Toxicology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a method for rapidly screening bacteriocin-producing microorganisms, which comprises the following steps of 1) under the aseptic condition, taking a sample to be tested, grinding the sample, then proportionally mixing the ground sample into physiological saline, and standing the mixture at 37 ℃ for later use; 2) taking 100 mu L of sample subjected to gradient dilution and respectively coating the sample on nutrient agar plates; 3) respectively carrying out anaerobic culture and aerobic culture on the coated flat plate at the temperature of 37 ℃; 4) adding 100 mu L of overnight cultured indicator bacteria into soft agar according to the proportion of 0.4-0.8%, uniformly mixing, pouring into a flat plate, and culturing at 37 ℃; 5) observing an inhibition zone of the cultured plate A, inoculating bacteria producing antibacterial substances by using an aseptic inoculating needle, and screening out a sample to be tested producing bacteriocin; the invention mixes all pathogenic microorganisms as indicator bacteria, adds the indicator bacteria into the soft agar, and realizes the rapid screening work of the bacteriocin-producing microorganisms by observing the inhibition zone.
Description
Technical Field
The invention pertains to microorganisms or enzymes; the technical field of the composition thereof, in particular to a method for rapidly screening a bacteriocin-producing microorganism.
Background
Bacteriocin is a polypeptide which is synthesized by ribosome of bacteria and has high-efficiency inhibition effect on relative bacteria; is generally considered to be one of bacteriostatic substances replacing antibiotics; at present, nisin (nisin) is the only bacteriocin authorized by WHO/FDA and allowed to be used in food, and the main reason is that nisin has broad-spectrum bacteriostatic effect, and related researches report the bacteriostatic mechanism of nisin in detail; however, as nisin-resistant pathogenic microorganisms are continuously detected, it is of great significance to screen novel bacteriocins which have broad spectrum and high bacteriostatic effect on pathogenic microorganisms.
Screening for bacteriocin-producing microorganisms is the first and important step in the discovery and utilization of bacteriocins, the most widely used screening method being the agar diffusion method; the agar diffusion method has the advantages of simple operation, visual result and the like, and has the defects of poor accuracy, poor repeatability and the like, so that the agar diffusion method is difficult to be used as a high-throughput screening method; other methods, such as microplate bioassay, bioluminescence, PCR rapid detection, etc., have great requirements for operation, and in addition, these methods are difficult to achieve for rapid screening.
Disclosure of Invention
In view of the above, in order to overcome the above-mentioned defects of the prior art, the present invention aims to provide a method for rapidly screening a bacteriocin-producing microorganism, wherein a sample is homogenized, plated and cultured; after the culture is finished, adding different pathogenic microorganisms into soft agar (0.6 percent of agar addition), directly pouring the mixture onto a plate full of bacteria, and culturing again; the pathogenic microorganism with bacteriocin inhibition effect is utilized to generate the effect of a bacteriostatic zone, and the aim of rapidly screening the bacteriocin-producing microorganism is fulfilled.
In order to achieve the above object, the present invention provides a method for rapidly screening a bacteriocin-producing microorganism, comprising the steps of:
1) under the aseptic condition, grinding a sample to be detected, then proportionally mixing the ground sample into physiological saline, and standing the mixture for later use at 37 ℃;
2) taking 100 mu L of sample subjected to gradient dilution and respectively coating the sample on nutrient agar plates;
3) respectively carrying out anaerobic culture and aerobic culture on the coated flat plate at the temperature of 37 ℃;
4) adding 100 mu L of overnight cultured indicator bacteria into soft agar according to the proportion of 0.4-0.8%, uniformly mixing, pouring into a flat plate, and culturing at 37 ℃;
5) observing the bacteriostasis zone of the cultured A plate, inoculating bacteria producing bacteriostasis substances by using an aseptic inoculating needle, and screening out a sample to be tested producing bacteriocin.
Further, the indicator bacteria are set to listeria monocytogenes, bacillus cereus and staphylococcus aureus.
Further, in the step 1), the ratio of the sample to be detected to the physiological saline is set to be 1: 8-1: 10.
further, the ratio of the sample to be detected to the normal saline is set to be 1: 8-1: 10.
further, the sample dilution ratio was set to 1:500 and 1: 800.
Further, soft agar was added in a proportion of 0.6%.
Compared with the prior art, the invention has the following beneficial effects:
1. the method for rapidly screening the bacteriocin-producing microorganisms comprises the steps of homogenizing a sample, coating a plate, and culturing; after the culture is finished, adding different pathogenic microorganisms into soft agar (0.6 percent of agar addition), directly pouring the mixture onto a plate full of bacteria, and culturing again; the pathogenic microorganism with bacteriocin inhibition effect is utilized to generate the effect of a bacteriostatic zone, so that the aim of rapidly screening the bacteriocin-producing microorganism is fulfilled;
2. the method for rapidly screening the bacteriocin-producing microorganisms can completely realize rapid screening of high-throughput, full-automatic and ready-to-use bacteriocin-producing microorganisms; the screening work from the sample to the bacteriocin-producing microorganism can be completely realized within 30h in the whole process; saves time and money for the development of the subsequent work of separating and extracting the bacteriocin.
Drawings
In order to make the object, technical scheme and beneficial effect of the invention more clear, the invention provides the following drawings for explanation:
FIG. 1 is a diagram showing the results of screening for listeria monocytogenes-inhibiting strains according to the present invention;
FIG. 2 is a graph showing the results of the screening for inhibition of Bacillus cereus of the present invention;
FIG. 3 is a graph showing the results of screening for inhibition of Staphylococcus aureus in accordance with the present invention;
FIG. 4 is a graph showing the results of the present invention simultaneously inhibiting Listeria monocytogenes, Bacillus cereus and Staphylococcus aureus.
Detailed Description
Preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings.
The invention relates to a method for rapidly screening listeria monocytogenes microorganisms, which comprises the following implementation modes:
example one
1) Under the aseptic condition, taking 25g of a sample to be detected, grinding, then proportionally mixing into 225mL of physiological saline, and standing at 37 ℃ for later use;
2) respectively coating 100 mu L of diluted samples which are 50 times, 100 times, 200 times, 500 times, 800 times and 1600 times on nutrient agar plates;
3) respectively carrying out anaerobic culture and aerobic culture on the coated flat plate at the temperature of 37 ℃;
4) adding 100 mu L of overnight cultured Listeria monocytogenes into 5mL of soft agar according to the proportion of 0.6 percent, uniformly mixing, pouring into a flat plate, and culturing at 37 ℃;
5) observing the bacteriostasis zone of the cultured A plate, inoculating bacteria producing bacteriostasis substances by using an aseptic inoculating needle, and screening out a sample to be tested producing bacteriocin.
Example two
1) Under the aseptic condition, taking 25g of a sample to be detected, grinding, then proportionally mixing into 175mL of physiological saline, and standing at 37 ℃ for later use;
2) respectively coating 100 mu L of diluted samples which are 50 times, 100 times, 200 times, 500 times, 800 times and 1600 times on nutrient agar plates;
3) respectively carrying out anaerobic culture and aerobic culture on the coated flat plate at the temperature of 37 ℃;
4) adding 100 mu L of overnight cultured Listeria monocytogenes into 5mL of soft agar according to the proportion of 0.4%, uniformly mixing, pouring into a flat plate, and culturing at 37 ℃;
5) observing the inhibition zone of the cultured plate A, inoculating bacteria producing antibacterial substances by using an aseptic inoculating needle as shown in figure 1, and screening out a sample to be tested producing bacteriocin.
EXAMPLE III
1) Under the aseptic condition, taking 25g of a sample to be detected, grinding, then proportionally mixing into 250mL of physiological saline, and standing at 37 ℃ for later use;
2) respectively coating 100 mu L of diluted samples which are 50 times, 100 times, 200 times, 500 times, 800 times and 1600 times on nutrient agar plates;
3) respectively carrying out anaerobic culture and aerobic culture on the coated flat plate at the temperature of 37 ℃;
4) adding 100 mu L of overnight cultured Listeria monocytogenes into 5mL of soft agar according to the proportion of 0.8 percent, uniformly mixing, pouring into a flat plate, and culturing at 37 ℃;
5) observing the bacteriostasis zone of the cultured A plate, inoculating bacteria producing bacteriostasis substances by using an aseptic inoculating needle, and screening out a sample to be tested producing bacteriocin.
FIG. 1 shows the effect of screening for microorganisms inhibiting Listeria monocytogenes.
The observed effect of inhibition cycles number for screening microorganisms inhibiting listeria monocytogenes in the above examples is shown in the following table:
the invention relates to a method for rapidly screening microorganisms producing bacillus cereus; the implementation mode is as follows:
example four
1) Under the aseptic condition, taking 25g of a sample to be detected, grinding, then proportionally mixing into 225mL of physiological saline, and standing at 37 ℃ for later use;
2) respectively coating 100 mu L of diluted samples which are 50 times, 100 times, 200 times, 500 times, 800 times and 1600 times on nutrient agar plates;
3) respectively carrying out anaerobic culture and aerobic culture on the coated flat plate at the temperature of 37 ℃;
4) adding 100 mu L of overnight cultured bacillus cereus into 5mL of soft agar according to the proportion of 0.6%, uniformly mixing, pouring into a flat plate, and culturing at 37 ℃;
5) observing the bacteriostasis zone of the cultured A plate, inoculating bacteria producing bacteriostasis substances by using an aseptic inoculating needle, and screening out a sample to be tested producing bacteriocin.
EXAMPLE five
1) Under the aseptic condition, taking 25g of a sample to be detected, grinding, then proportionally mixing into 175mL of physiological saline, and standing at 37 ℃ for later use;
2) respectively coating 100 mu L of diluted samples which are 50 times, 100 times, 200 times, 500 times, 800 times and 1600 times on nutrient agar plates;
3) respectively carrying out anaerobic culture and aerobic culture on the coated flat plate at the temperature of 37 ℃;
4) adding 100 mu L of overnight cultured bacillus cereus into 5mL of soft agar according to the proportion of 0.4%, uniformly mixing, pouring into a flat plate, and culturing at 37 ℃;
5) observing the inhibition zone of the cultured plate A, inoculating bacteria producing antibacterial substances by using an aseptic inoculating needle as shown in figure 1, and screening out a sample to be tested producing bacteriocin.
EXAMPLE six
1) Under the aseptic condition, taking 25g of a sample to be detected, grinding, then proportionally mixing into 250mL of physiological saline, and standing at 37 ℃ for later use;
2) respectively coating 100 mu L of diluted samples which are 50 times, 100 times, 200 times, 500 times, 800 times and 1600 times on nutrient agar plates;
3) respectively carrying out anaerobic culture and aerobic culture on the coated flat plate at the temperature of 37 ℃;
4) adding 100 mu L of overnight cultured bacillus cereus into 5mL of soft agar according to the proportion of 0.8%, uniformly mixing, pouring into a flat plate, and culturing at 37 ℃;
5) observing the bacteriostasis zone of the cultured A plate, inoculating bacteria producing bacteriostasis substances by using an aseptic inoculating needle, and screening out a sample to be tested producing bacteriocin.
FIG. 2 shows the effect of screening for microorganisms inhibiting Bacillus cereus.
The observed inhibition cycle number effect for screening microorganisms capable of inhibiting bacillus cereus in the above example is shown in the following table:
the invention relates to a method for rapidly screening staphylococcus aureus microorganisms; the implementation mode is as follows:
EXAMPLE seven
1) Under the aseptic condition, taking 25g of a sample to be detected, grinding, then proportionally mixing into 225mL of physiological saline, and standing at 37 ℃ for later use;
2) respectively coating 100 mu L of diluted samples which are 50 times, 100 times, 200 times, 500 times, 800 times and 1600 times on nutrient agar plates;
3) respectively carrying out anaerobic culture and aerobic culture on the coated flat plate at the temperature of 37 ℃;
4) adding 100 mu L of overnight cultured staphylococcus aureus into 5mL of soft agar according to the proportion of 0.6 percent, uniformly mixing, pouring into a flat plate, and culturing at 37 ℃;
5) observing the bacteriostasis zone of the cultured A plate, inoculating bacteria producing bacteriostasis substances by using an aseptic inoculating needle, and screening out a sample to be tested producing bacteriocin.
Example eight
1) Under the aseptic condition, taking 25g of a sample to be detected, grinding, then proportionally mixing into 175mL of physiological saline, and standing at 37 ℃ for later use;
2) respectively coating 100 mu L of diluted samples which are 50 times, 100 times, 200 times, 500 times, 800 times and 1600 times on nutrient agar plates;
3) respectively carrying out anaerobic culture and aerobic culture on the coated flat plate at the temperature of 37 ℃;
4) adding 100 mu L of overnight cultured staphylococcus aureus into 5mL of soft agar according to the proportion of 0.4%, uniformly mixing, pouring into a flat plate, and culturing at 37 ℃;
5) observing the inhibition zone of the cultured plate A, inoculating bacteria producing antibacterial substances by using an aseptic inoculating needle as shown in figure 1, and screening out a sample to be tested producing bacteriocin.
Example nine
1) Under the aseptic condition, taking 25g of a sample to be detected, grinding, then proportionally mixing into 250mL of physiological saline, and standing at 37 ℃ for later use;
2) respectively coating 100 mu L of diluted samples which are 50 times, 100 times, 200 times, 500 times, 800 times and 1600 times on nutrient agar plates;
3) respectively carrying out anaerobic culture and aerobic culture on the coated flat plate at the temperature of 37 ℃;
4) adding 100 mu L of overnight cultured staphylococcus aureus into 5mL of soft agar according to the proportion of 0.8 percent, uniformly mixing, pouring into a flat plate, and culturing at 37 ℃;
5) observing the bacteriostasis zone of the cultured A plate, inoculating bacteria producing bacteriostasis substances by using an aseptic inoculating needle, and screening out a sample to be tested producing bacteriocin.
FIG. 3 shows the effect of screening microorganisms inhibiting Staphylococcus aureus.
The observed effect of inhibition cycles for screening microorganisms inhibiting staphylococcus aureus in the above example is shown in the following table:
example ten
This example screens microorganisms that simultaneously inhibit listeria monocytogenes, bacillus cereus and staphylococcus aureus as follows:
1) under the aseptic condition, taking 25g of a sample to be detected, grinding, then proportionally mixing into 225mL of physiological saline, and standing at 37 ℃ for later use;
2) respectively coating 100 mu L of diluted samples which are 50 times, 100 times, 200 times, 500 times, 800 times and 1600 times on nutrient agar plates;
3) respectively carrying out anaerobic culture and aerobic culture on the coated flat plate at the temperature of 37 ℃;
4) taking 100 mu L of overnight cultured mixed bacterium liquid of listeria monocytogenes, bacillus cereus and staphylococcus aureus, adding the mixed bacterium liquid into soft agar according to the proportion of 6 percent, uniformly mixing, pouring the mixture into a flat plate, and culturing at the temperature of 37 ℃;
5) observing the bacteriostatic zone of the cultured A plate, inoculating bacteria producing bacteriostatic substances by using an aseptic inoculating needle, and screening out a sample to be tested producing bacteriocin
FIG. 4 shows the effect of screening microorganisms capable of simultaneously inhibiting Listeria monocytogenes, Bacillus cereus and Staphylococcus aureus.
Finally, it is noted that the above-mentioned preferred embodiments illustrate rather than limit the invention, and that, although the invention has been described in detail with reference to the above-mentioned preferred embodiments, it will be understood by those skilled in the art that various changes in form and detail may be made therein without departing from the scope of the invention as defined by the appended claims.
Claims (6)
1. A method for rapidly screening a bacteriocin-producing microorganism, which is characterized by comprising the following steps: the method comprises the following steps:
1) under the aseptic condition, grinding a sample to be detected, then proportionally mixing the ground sample into physiological saline, and standing the mixture for later use at 37 ℃;
2) taking 100 mu L of sample subjected to gradient dilution and respectively coating the sample on nutrient agar plates;
3) respectively carrying out anaerobic culture and aerobic culture on the coated flat plate at the temperature of 37 ℃;
4) adding 100 mu L of overnight cultured indicator bacteria into soft agar according to the proportion of 0.4-0.8%, uniformly mixing, pouring into a flat plate, and culturing at 37 ℃;
5) observing the bacteriostasis zone of the cultured A plate, inoculating bacteria producing bacteriostasis substances by using an aseptic inoculating needle, and screening out a sample to be tested producing bacteriocin.
2. The method for rapid screening of a bacteriocin-producing microorganism according to claim 1, characterized in that: the indicator bacteria are set to Listeria monocytogenes, Bacillus cereus and Staphylococcus aureus.
3. The method for rapid screening of a bacteriocin-producing microorganism according to claim 1, characterized in that: in the step 1), the ratio of the sample to be detected to the normal saline is set to be 1: 8-1: 10.
4. the method for rapid screening of a bacteriocin-producing microorganism according to claim 3, characterized in that: the ratio of the sample to be detected to the normal saline is set as 1: 9.
5. the method for rapid screening of a bacteriocin-producing microorganism according to claim 3, characterized in that: the sample dilution ratios were set at 1:500 and 1: 800.
6. The method for rapid screening of a bacteriocin-producing microorganism according to claim 3, characterized in that: in step 4), soft agar was added in a proportion of 0.6%.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210068639.8A CN114395610A (en) | 2022-01-20 | 2022-01-20 | Method for rapidly screening bacteriocin-producing microorganisms |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210068639.8A CN114395610A (en) | 2022-01-20 | 2022-01-20 | Method for rapidly screening bacteriocin-producing microorganisms |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114395610A true CN114395610A (en) | 2022-04-26 |
Family
ID=81232210
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210068639.8A Pending CN114395610A (en) | 2022-01-20 | 2022-01-20 | Method for rapidly screening bacteriocin-producing microorganisms |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114395610A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20000021520A (en) * | 1998-09-30 | 2000-04-25 | 변유량 | Lactococcus lactics(kfcc-11047) producing natural antibacterial bacteriocin |
| CN102250806A (en) * | 2011-07-01 | 2011-11-23 | 安徽农业大学 | Method for screening lactic acid bacteria capable of producing bacteriocin from plant source materials |
| CN105400721A (en) * | 2015-12-11 | 2016-03-16 | 湖南农业大学 | Method for rapid screening of Bacillus spp with antibacterial activity |
| CN105462839A (en) * | 2016-01-18 | 2016-04-06 | 浙江大学 | Fast screening method of pathogenic-bacterium-resistant bacillus |
-
2022
- 2022-01-20 CN CN202210068639.8A patent/CN114395610A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20000021520A (en) * | 1998-09-30 | 2000-04-25 | 변유량 | Lactococcus lactics(kfcc-11047) producing natural antibacterial bacteriocin |
| CN102250806A (en) * | 2011-07-01 | 2011-11-23 | 安徽农业大学 | Method for screening lactic acid bacteria capable of producing bacteriocin from plant source materials |
| CN105400721A (en) * | 2015-12-11 | 2016-03-16 | 湖南农业大学 | Method for rapid screening of Bacillus spp with antibacterial activity |
| CN105462839A (en) * | 2016-01-18 | 2016-04-06 | 浙江大学 | Fast screening method of pathogenic-bacterium-resistant bacillus |
Non-Patent Citations (1)
| Title |
|---|
| 王娜等: "产细菌素乳酸菌的筛选与鉴定", 《中国食品学报》, vol. 20, no. 12, pages 248 - 255 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Matsumoto et al. | Endophytic actinomycetes: promising source of novel bioactive compounds | |
| Williams et al. | Use of antibiotics for selective isolation and enumeration of actinomycetes in soil | |
| Rutgersson et al. | Long-term application of Swedish sewage sludge on farmland does not cause clear changes in the soil bacterial resistome | |
| US20170267599A1 (en) | Method for preparing solid trichoderma seed from direct fermentation of crop straws with trichoderma, and product prepared by using the same | |
| CN1948535A (en) | Rare earth improving antibacterial performance silver containing stainless steel | |
| CN103642891A (en) | Method for inspecting microbes in cosmetics | |
| CN104211184A (en) | Microbial sewage treating agent used for livestock and poultry breeding wastewater treatment | |
| CN113444657A (en) | Solid-state microbial composite microbial agent for promoting aerobic composting of kitchen waste and preparation and application thereof | |
| CN106591416B (en) | Chromogenic medium for detecting food-borne pathogenic yersinia | |
| CN1295344C (en) | Screening method for phophonomycin biological conversion strain | |
| CN114395610A (en) | Method for rapidly screening bacteriocin-producing microorganisms | |
| Bhattacharya et al. | Lipase and protease producing microbes from the environment of sugar beet field | |
| CN114702141B (en) | Application of aniline degrading bacteria Proveus lanuginosus in dye decolorization | |
| Wilcockson et al. | Nitrogenase activity of Rhizobium japonicum growing on agar surfaces in relation to slime production, growth and survival | |
| Hudha et al. | Manufacture of local microorganism (MOL) from vegetable waste with nutrition source supply variation | |
| Yang et al. | Morphogenesis, ATP content and oxytetracycline production by Streptomyces rimosus in solid substrate cultivation | |
| Gill et al. | Clinical and microbiological observations on CDC group DF-3, a gram-negative coccobacillus | |
| Laslo et al. | The management of N and P nutrition of plants using nitrogen fixing and phosphorus solubilizing bacteria | |
| Fereidouni Velasejerdi et al. | Investigation of enzymes and solvents in the production process of 6-ammonium penicillanic acid (6-APA) in industry to reduce costs and improve production conditions | |
| Rastegari et al. | Existance of microbial species in vermicomposts derived from mixed sesame crust and cow manure treatments | |
| Rahimi et al. | Effects of medium and inoculum on antimicrobial susceptibility of anaerobic bacteria | |
| EP0281523A2 (en) | Pseudomonas | |
| ARP et al. | Comparison of detection speed and yield in agitated and non‐agitated aerobic blood culture bottles | |
| Blazevic | Evaluation of the modified broth-disk method for determining antibiotic susceptibilities of anaerobic bacteria | |
| CN113929221B (en) | Microorganism composite flora for treating aquaculture tail water |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |