CN110592174A - Antagonistic antibacterial component selective enrichment culture medium and preparation and use methods thereof - Google Patents
Antagonistic antibacterial component selective enrichment culture medium and preparation and use methods thereof Download PDFInfo
- Publication number
- CN110592174A CN110592174A CN201910933605.9A CN201910933605A CN110592174A CN 110592174 A CN110592174 A CN 110592174A CN 201910933605 A CN201910933605 A CN 201910933605A CN 110592174 A CN110592174 A CN 110592174A
- Authority
- CN
- China
- Prior art keywords
- selective enrichment
- resin
- enrichment medium
- antagonistic
- lithium chloride
- 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
- 230000003042 antagnostic effect Effects 0.000 title claims abstract description 31
- 230000000844 anti-bacterial effect Effects 0.000 title claims abstract description 21
- 238000000034 method Methods 0.000 title claims abstract description 9
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 239000001963 growth medium Substances 0.000 title claims description 14
- 229920005989 resin Polymers 0.000 claims abstract description 59
- 239000011347 resin Substances 0.000 claims abstract description 59
- KWGKDLIKAYFUFQ-UHFFFAOYSA-M lithium chloride Chemical compound [Li+].[Cl-] KWGKDLIKAYFUFQ-UHFFFAOYSA-M 0.000 claims abstract description 56
- DAEPDZWVDSPTHF-UHFFFAOYSA-M sodium pyruvate Chemical compound [Na+].CC(=O)C([O-])=O DAEPDZWVDSPTHF-UHFFFAOYSA-M 0.000 claims abstract description 44
- 239000002609 medium Substances 0.000 claims abstract description 38
- NGVDGCNFYWLIFO-UHFFFAOYSA-N pyridoxal 5'-phosphate Chemical compound CC1=NC=C(COP(O)(O)=O)C(C=O)=C1O NGVDGCNFYWLIFO-UHFFFAOYSA-N 0.000 claims abstract description 32
- 241000894006 Bacteria Species 0.000 claims abstract description 31
- 241000186779 Listeria monocytogenes Species 0.000 claims abstract description 26
- 241000191967 Staphylococcus aureus Species 0.000 claims abstract description 26
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 22
- 239000008103 glucose Substances 0.000 claims abstract description 22
- 229940054269 sodium pyruvate Drugs 0.000 claims abstract description 22
- 241000607142 Salmonella Species 0.000 claims abstract description 20
- 241000607768 Shigella Species 0.000 claims abstract description 20
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 20
- 239000003112 inhibitor Substances 0.000 claims abstract description 20
- FRHBOQMZUOWXQL-UHFFFAOYSA-L ammonium ferric citrate Chemical compound [NH4+].[Fe+3].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O FRHBOQMZUOWXQL-UHFFFAOYSA-L 0.000 claims abstract description 19
- 235000000011 iron ammonium citrate Nutrition 0.000 claims abstract description 19
- 239000004313 iron ammonium citrate Substances 0.000 claims abstract description 19
- BFPJYWDBBLZXOM-UHFFFAOYSA-L potassium tellurite Chemical compound [K+].[K+].[O-][Te]([O-])=O BFPJYWDBBLZXOM-UHFFFAOYSA-L 0.000 claims abstract description 19
- 229960003964 deoxycholic acid Drugs 0.000 claims abstract description 17
- KXGVEGMKQFWNSR-LLQZFEROSA-N deoxycholic acid Chemical compound C([C@H]1CC2)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 KXGVEGMKQFWNSR-LLQZFEROSA-N 0.000 claims abstract description 17
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 claims abstract description 16
- 235000007682 pyridoxal 5'-phosphate Nutrition 0.000 claims abstract description 16
- 239000011589 pyridoxal 5'-phosphate Substances 0.000 claims abstract description 16
- 229960001327 pyridoxal phosphate Drugs 0.000 claims abstract description 16
- 238000001179 sorption measurement Methods 0.000 claims abstract description 16
- 125000002091 cationic group Chemical group 0.000 claims abstract description 13
- 229930003779 Vitamin B12 Natural products 0.000 claims abstract description 11
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims abstract description 11
- FDJOLVPMNUYSCM-WZHZPDAFSA-L cobalt(3+);[(2r,3s,4r,5s)-5-(5,6-dimethylbenzimidazol-1-yl)-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl] [(2r)-1-[3-[(1r,2r,3r,4z,7s,9z,12s,13s,14z,17s,18s,19r)-2,13,18-tris(2-amino-2-oxoethyl)-7,12,17-tris(3-amino-3-oxopropyl)-3,5,8,8,13,15,18,19-octamethyl-2 Chemical compound [Co+3].N#[C-].N([C@@H]([C@]1(C)[N-]\C([C@H]([C@@]1(CC(N)=O)C)CCC(N)=O)=C(\C)/C1=N/C([C@H]([C@@]1(CC(N)=O)C)CCC(N)=O)=C\C1=N\C([C@H](C1(C)C)CCC(N)=O)=C/1C)[C@@H]2CC(N)=O)=C\1[C@]2(C)CCC(=O)NC[C@@H](C)OP([O-])(=O)O[C@H]1[C@@H](O)[C@@H](N2C3=CC(C)=C(C)C=C3N=C2)O[C@@H]1CO FDJOLVPMNUYSCM-WZHZPDAFSA-L 0.000 claims abstract description 11
- 239000011715 vitamin B12 Substances 0.000 claims abstract description 11
- 235000019163 vitamin B12 Nutrition 0.000 claims abstract description 11
- 230000003385 bacteriostatic effect Effects 0.000 claims abstract description 10
- 229960004642 ferric ammonium citrate Drugs 0.000 claims abstract description 10
- 108010050327 trypticase-soy broth Proteins 0.000 claims abstract description 9
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 claims abstract description 8
- 229960002743 glutamine Drugs 0.000 claims abstract description 8
- -1 malt extract Substances 0.000 claims abstract description 8
- 239000011780 sodium chloride Substances 0.000 claims abstract description 8
- 235000002639 sodium chloride Nutrition 0.000 claims abstract description 8
- 239000001974 tryptic soy broth Substances 0.000 claims abstract description 8
- 229960001031 glucose Drugs 0.000 claims abstract description 5
- 229940045184 malt extract Drugs 0.000 claims abstract description 5
- 229920001429 chelating resin Polymers 0.000 claims description 24
- 239000012153 distilled water Substances 0.000 claims description 24
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 24
- 101100313763 Arabidopsis thaliana TIM22-2 gene Proteins 0.000 claims description 9
- 239000003463 adsorbent Substances 0.000 claims description 8
- 239000007864 aqueous solution Substances 0.000 claims description 8
- 244000005700 microbiome Species 0.000 claims description 8
- 239000002872 contrast media Substances 0.000 claims description 6
- 239000002245 particle Substances 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 5
- 229940088594 vitamin Drugs 0.000 claims description 5
- 229930003231 vitamin Natural products 0.000 claims description 5
- 235000013343 vitamin Nutrition 0.000 claims description 5
- 239000011782 vitamin Substances 0.000 claims description 5
- 150000003722 vitamin derivatives Chemical class 0.000 claims description 5
- 239000000243 solution Substances 0.000 claims description 4
- 238000010790 dilution Methods 0.000 claims description 3
- 239000012895 dilution Substances 0.000 claims description 3
- 238000002474 experimental method Methods 0.000 claims description 3
- 235000004554 glutamine Nutrition 0.000 claims description 3
- 229940073577 lithium chloride Drugs 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 230000003115 biocidal effect Effects 0.000 abstract description 3
- 235000010469 Glycine max Nutrition 0.000 description 10
- 241000588724 Escherichia coli Species 0.000 description 7
- 241000607272 Vibrio parahaemolyticus Species 0.000 description 7
- 241001354013 Salmonella enterica subsp. enterica serovar Enteritidis Species 0.000 description 6
- 241000607762 Shigella flexneri Species 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 238000001514 detection method Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 4
- 230000002401 inhibitory effect Effects 0.000 description 4
- 229920001817 Agar Polymers 0.000 description 3
- 239000008272 agar Substances 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 235000013305 food Nutrition 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 229940099352 cholate Drugs 0.000 description 2
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 230000001965 increasing effect Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 244000000010 microbial pathogen Species 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 241000193755 Bacillus cereus Species 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- 241000607447 Yersinia enterocolitica Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009640 blood culture Methods 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 239000012913 medium supplement Substances 0.000 description 1
- 239000002068 microbial inoculum Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 229940098232 yersinia enterocolitica Drugs 0.000 description 1
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
- C12Q1/045—Culture media therefor
-
- 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
- C12Q1/10—Enterobacteria
-
- 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
- C12Q1/14—Streptococcus; Staphylococcus
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/24—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
- G01N2333/25—Shigella (G)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/24—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
- G01N2333/255—Salmonella (G)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/305—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Micrococcaceae (F)
- G01N2333/31—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Micrococcaceae (F) from Staphylococcus (G)
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Toxicology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses an antagonistic antibacterial component selective enrichment medium and a preparation and use method thereof. The invention can adsorb various antibiotic and other bacteriostatic components through macroporous adsorption resin or cationic exchange resin, the inhibitor is prepared from lithium chloride, potassium tellurite, sodium deoxycholate and ferric ammonium citrate to inhibit the growth of non-target bacteria, the promoter is prepared from trypticase soy broth, malt extract, glucose, glutamine, sodium pyruvate, pyridoxal phosphate, vitamin B12 and sodium chloride to enable the target bacteria, namely salmonella, staphylococcus aureus, shigella and listeria monocytogenes to grow rapidly, the selective enrichment medium products for salmonella, staphylococcus aureus, shigella and listeria monocytogenes on the market are supplemented, and the blank of the selective enrichment medium market for adsorbing the bacteriostatic components is filled.
Description
Technical Field
The invention relates to the technical field of culture of pathogenic bacteria in food, in particular to a selective enrichment medium for antagonistic antibacterial components and a preparation and use method thereof.
Background
Microorganisms in samples for food safety detection are often subjected to physical damage in the transfer process, chemical damage of residual drugs and other bacteriostatic chemical components, and the pathogenic microorganisms are easily shocked, such as a false death state, so that the detection cannot be detected or the result is false negative, and the hidden danger of harming the public health is formed. The three links of restoration of thallus cell wall damage, absorption of bacteriostatic components and selective rapid enrichment can be considered to be linked together for integrated research, and a rapid enrichment medium for antagonizing the bacteriostatic components is provided for various detection methods (blood culture bottles) of pathogenic microorganisms.
Disclosure of Invention
The invention mainly aims to provide an antagonistic antibacterial component selective enrichment culture medium and a preparation and use method thereof, which can effectively solve the problems in the background technology.
In order to achieve the purpose, the invention adopts the technical scheme that:
a selective enrichment medium for antagonistic antibacterial components comprises resin, inhibitor and promoter, wherein the medium is a mixture of the resin, the inhibitor and the promoter.
Further, the resin is any one of Amberlyst A21 macroporous adsorption resin, DIAION HP20 macroporous adsorption resin and Amberlite IRC-84 cationic exchange resin, the Amberlyst A21 macroporous adsorption resin is preferably Amberlyst A21 macroporous adsorption resin of Merck pharmaceutical biotechnology, Germany, the Amberlyst is a Luomax resin series standard, the DIAION HP20 macroporous adsorption resin is preferably DIAION NHP20 macroporous adsorption resin of Mitsubishi chemical company, Japan, and the Amberlite IRC-84 cationic exchange resin is preferably Amberlite IRC-84 cationic exchange resin of Dow chemical company, USA.
Further, the inhibitor is one or more of lithium chloride, potassium tellurite, sodium deoxycholate and ferric ammonium citrate.
Further, the promoter is one or more selected from trypticase soy broth, malt extract, glucose, glutamine, sodium pyruvate, pyridoxal phosphate, vitamin B12 and sodium chloride.
Further, the lithium chloride, the potassium tellurite, the sodium deoxycholate and the ferric ammonium citrate are powdery particles.
Further, the trypticase soy broth, malt extract, glucose and sodium pyruvate are in the form of powdered particles, and the lithium chloride, glutamine, pyridoxal phosphate and vitamin B12 are in the form of aqueous solutions.
A preparation method of a selective enrichment medium for antagonistic antibacterial components comprises the following steps:
step 1: preparing a certain amount of distilled water, taking out lithium chloride, potassium tellurite, sodium deoxycholate and ammonium ferric citrate, pouring the lithium chloride, the potassium tellurite, the sodium deoxycholate and the ammonium ferric citrate into the distilled water step by step or at normal temperature and normal pressure, and fully stirring to dissolve the lithium chloride, the potassium tellurite, the sodium deoxycholate and the ammonium ferric citrate into the distilled water to form the inhibitor.
Step 2: preparing a certain amount of distilled water again, taking out tryptose soy broth, malt extract, glucose and sodium pyruvate, pouring the tryptose soy broth, the malt extract, the glucose and the sodium pyruvate into the distilled water gradually or a plurality of the tryptose soy broth, the malt extract, the glucose and the sodium pyruvate under normal temperature and normal pressure, fully stirring to dissolve the tryptose soy broth, the malt extract, the glucose and the sodium pyruvate into the distilled water to form an accelerating solution, filtering the lithium chloride, the glutamine, the pyridoxal phosphate and the vitamin B12 aqueous solution gradually or a plurality of the lithium chloride, the glutamine, the pyridoxal phosphate and the vitamin.
And step 3: a quantity of Amberlyst A21 macroporous adsorbent resin, DIAION HP20 macroporous adsorbent resin and Amberlite IRC-84 cationic exchange resin was used as resin.
And 4, step 4: and pouring the inhibitor and the accelerant into resin to form the selective enrichment medium for the antagonistic antibacterial components.
A use method of a selective enrichment medium for antagonistic antibacterial components comprises the following steps:
step 1: salmonella, staphylococcus aureus, shigella and Listeria monocytogenes are respectively inoculated on the surface of the selective enrichment medium for antagonistic bacteriostatic components in an inclined manner, and the surface of the selective enrichment medium is subjected to gradient dilution by sterile normal saline.
Step 2: and placing the antagonistic antibacterial component selective enrichment culture medium inoculated with salmonella, staphylococcus aureus, shigella and listeria monocytogenes into a thermostat suitable for the temperature required by the growth of microorganisms for shake culture at 180 r/min.
And step 3: inoculating salmonella, staphylococcus aureus, shigella and listeria monocytogenes on resin as a contrast medium, and placing the contrast medium in a thermostat with a temperature suitable for the growth of microorganisms to carry out shake culture at 180 r/min.
And 4, step 4: the two are simultaneously shake-cultured for 24 hours; the cultures of 4h, 8h, 12h, 16h, 20h and 24h are respectively counted, and the parallel experiments are carried out for 10 times, and the thallus concentrations at different times are compared.
Compared with the prior art, the invention has the following beneficial effects:
(1) the invention can adsorb various antibiotic and other bacteriostatic components through macroporous adsorption resin or cationic exchange resin.
(2) The inhibitor is prepared from lithium chloride, potassium tellurite, sodium deoxycholate and ferric ammonium citrate to inhibit the growth of non-target bacteria.
(3) The promoter is prepared from trypticase soy broth, malt extract, glucose, glutamine, sodium pyruvate, pyridoxal phosphate, vitamin B12 and sodium chloride, so that target bacteria, namely salmonella, staphylococcus aureus, shigella and listeria monocytogenes, can grow quickly.
(4) The selective enrichment medium supplements the selective enrichment medium products for salmonella, staphylococcus aureus, shigella and listeria monocytogenes on the market, and fills the blank of the selective enrichment medium market for adsorbing bacteriostatic components.
Detailed Description
In order to make the technical means, the creation characteristics, the achievement purposes and the effects of the invention easy to understand, the invention is further described with the specific embodiments.
As shown in the following, the selective enrichment medium for the antagonistic antibacterial components comprises resin, an inhibitor and an accelerant, and the medium is a mixture of the resin, the inhibitor and the accelerant.
Wherein the resin is any one of Amberlyst A21 macroporous adsorption resin, DIAION HP20 macroporous adsorption resin and Amberlite IRC-84 cationic exchange resin, the Amberlyst A21 macroporous adsorption resin is preferably Amberlyst A21 macroporous adsorption resin of Merck pharmaceutical and biotechnology, Germany, the Amberlyst is a Luomax resin series standard, the DIAION HP20 macroporous adsorption resin is preferably DIAION NHP20 macroporous adsorption resin of Mitsubishi chemical company, Japan, and the Amberlite IRC-84 cationic exchange resin is preferably Amberlite IRC-84 cationic exchange resin of Dow chemical company, USA.
Wherein the inhibitor is one or more of lithium chloride, potassium tellurite, sodium deoxycholate and ferric ammonium citrate.
Wherein the promoter is one or more of trypticase soy broth, malt extract, glucose, glutamine, sodium pyruvate, pyridoxal phosphate, vitamin B12 and sodium chloride.
Wherein the lithium chloride, the potassium tellurite, the sodium deoxycholate and the ferric ammonium citrate are powdery particles.
Wherein the trypticase soy broth, malt extract, glucose and sodium pyruvate are in the form of powdered particles, and the lithium chloride, glutamine, pyridoxal phosphate and vitamin B12 are in the form of aqueous solutions.
A preparation method of a selective enrichment medium for antagonistic antibacterial components comprises the following steps:
step 1: preparing a certain amount of distilled water, taking out lithium chloride, potassium tellurite, sodium deoxycholate and ammonium ferric citrate, pouring the lithium chloride, the potassium tellurite, the sodium deoxycholate and the ammonium ferric citrate into the distilled water step by step or at normal temperature and normal pressure, and fully stirring to dissolve the lithium chloride, the potassium tellurite, the sodium deoxycholate and the ammonium ferric citrate into the distilled water to form the inhibitor.
Step 2: preparing a certain amount of distilled water again, taking out tryptose soy broth, malt extract, glucose and sodium pyruvate, pouring the tryptose soy broth, the malt extract, the glucose and the sodium pyruvate into the distilled water gradually or a plurality of the tryptose soy broth, the malt extract, the glucose and the sodium pyruvate under normal temperature and normal pressure, fully stirring to dissolve the tryptose soy broth, the malt extract, the glucose and the sodium pyruvate into the distilled water to form an accelerating solution, filtering the lithium chloride, the glutamine, the pyridoxal phosphate and the vitamin B12 aqueous solution gradually or a plurality of the lithium chloride, the glutamine, the pyridoxal phosphate and the vitamin.
And step 3: a quantity of Amberlyst A21 macroporous adsorbent resin, DIAION HP20 macroporous adsorbent resin and Amberlite IRC-84 cationic exchange resin was used as resin.
And 4, step 4: and pouring the inhibitor and the accelerant into resin to form the selective enrichment medium for the antagonistic antibacterial components.
A use method of a selective enrichment medium for antagonistic antibacterial components comprises the following steps:
step 1: salmonella, staphylococcus aureus, shigella and Listeria monocytogenes are respectively inoculated on the surface of the selective enrichment medium for antagonistic bacteriostatic components in an inclined manner, and the surface of the selective enrichment medium is subjected to gradient dilution by sterile normal saline.
Step 2: and placing the antagonistic antibacterial component selective enrichment culture medium inoculated with salmonella, staphylococcus aureus, shigella and listeria monocytogenes into a thermostat suitable for the temperature required by the growth of microorganisms for shake culture at 180 r/min.
And step 3: inoculating salmonella, staphylococcus aureus, shigella and listeria monocytogenes on resin as a contrast medium, and placing the contrast medium in a thermostat with a temperature suitable for the growth of microorganisms to carry out shake culture at 180 r/min.
And 4, step 4: the two are simultaneously shake-cultured for 24 hours; the cultures of 4h, 8h, 12h, 16h, 20h and 24h are respectively counted, and the parallel experiments are carried out for 10 times, and the thallus concentrations at different times are compared.
It is to be noted that the invention relates to a selective enrichment medium for antagonistic bacteria-inhibiting components,
example 1
Salmonella enteritidis, staphylococcus aureus, Shigella flexneri, Listeria monocytogenes, Escherichia coli, Vibrio parahaemolyticus, Salmonella, staphylococcus aureus, Shigella and Listeria monocytogenes are selected as the experimental strains.
(1) 1000mL of distilled water was taken out.
(2) 3.0g of beef extract, 10.0g of peptone and 5.0g of sodium chloride are weighed, poured into distilled water and stirred uniformly.
(3) Adjusting pH to 7.4, sterilizing, and preserving at 4 deg.C in NB medium.
(4) 100mL of NB medium was taken out, 4 target bacteria and 6 non-target bacteria were inoculated into NB medium at an initial volume of 102CFU/mL, and shake-cultured at 37 ℃ and 180r/min for 24 hours. The cultures of 4h, 8h, 12h, 16h, 20h and 24h were counted by plate counting agar, and the cell concentration was compared at different times in parallel 10 times.
(5) The results show that the compound has no inhibition effect on 6 non-target bacteria such as salmonella enteritidis, staphylococcus aureus, shigella flexneri, listeria monocytogenes, escherichia coli and vibrio parahaemolyticus, and all grow normally, and the salmonella, the staphylococcus aureus, the shigella and the listeria monocytogenes grow normally.
Example 2
Salmonella enteritidis, staphylococcus aureus, Shigella flexneri, Listeria monocytogenes, Escherichia coli, Vibrio parahaemolyticus, Salmonella, staphylococcus aureus, Shigella and Listeria monocytogenes are selected as the experimental strains.
(1) 1000mL of distilled water was taken out.
(2) Selecting components in an inhibitor and an accelerator, weighing 0.2mg of potassium tellurite, 0.1g of deoxysodium cholate, 0.5g of ferric ammonium citrate, 30.0g of trypticase soybean broth, 5.0g of malt extract, 2.5g of glucose, 4.0g of sodium pyruvate, and 10g of sodium chloride, directly dissolving in distilled water, filtering an aqueous solution of 0.5g of lithium chloride, 0.5g of glutamine, 1.0mg of pyridoxal phosphate and 1.0mg of vitamin B121.0mg by using a 0.22-micron filter membrane, and then mixing with distilled water to form a culture medium, wherein the final volume of the culture medium is ensured to be 1000 mL.
(3) Taking out 100mL of culture medium, inoculating 4 kinds of target bacteria and 6 kinds of non-target bacteria into the culture medium according to the initial amount of 102CFU/mL, transferring for 2 times, diluting with sterile normal saline in a gradient manner, and performing shake culture at 37 ℃ and 180r/min for 24 hours. The cultures of 4h, 8h, 12h, 16h, 20h and 24h were counted by plate counting agar, and the cell concentration was compared at different times in parallel 10 times.
(4) The results show that the compound microbial inoculum has a certain inhibiting effect on 6 non-target bacteria such as salmonella enteritidis, staphylococcus aureus, shigella flexneri, listeria monocytogenes, escherichia coli and vibrio parahaemolyticus, the growth of all the non-target bacteria is slow, part of the non-target bacteria still grow after 20 hours, and the salmonella, the staphylococcus aureus, the shigella and the listeria monocytogenes are stable in growth, and have obvious bacterium increasing speed and bacterium increasing effect.
Example 3
Salmonella enteritidis, staphylococcus aureus, Shigella flexneri, Listeria monocytogenes, Escherichia coli, Vibrio parahaemolyticus, Salmonella, staphylococcus aureus, Shigella and Listeria monocytogenes are selected as the experimental strains.
(1) 1000mL of one of Amberlyst A21 macroporous adsorbent resin, DIAION HP20 macroporous adsorbent resin, and Amberlite IRC-84 cationic exchange resin was removed.
(2) Selecting components in an inhibitor and an accelerator, weighing 0.2mg of potassium tellurite, 0.1g of deoxysodium cholate, 0.5g of ferric ammonium citrate, 30.0g of tryptose soy broth, 5.0g of malt extract, 2.5g of glucose, 4.0g of sodium pyruvate and 10g of sodium chloride, directly dissolving in distilled water, filtering an aqueous solution of 0.5g of lithium chloride, 0.5g of glutamine, 1.0mg of pyridoxal phosphate and 1.0mg of vitamin B121.0mg by using a 0.22-micron filter membrane, and mixing into resin to form a SSSL culture medium, wherein the final volume of the SSSL culture medium is ensured to be 1000 mL.
(3) Taking 100mL of SSSL culture medium, inoculating 4 kinds of target bacteria and 6 kinds of non-target bacteria into the SSSL culture medium according to the initial amount of 102CFU/mL, transferring for 2 times, diluting with sterile physiological saline in a gradient manner, and culturing for 24 hours at 37 ℃ at 180r/min in a shaking environment. The cultures of 4h, 8h, 12h, 16h, 20h and 24h were counted by plate counting agar, and the cell concentration was compared at different times in parallel 10 times.
(4) The result shows that the compound bacterial preparation has obvious inhibition effect on 6 non-target bacteria of salmonella enteritidis, staphylococcus aureus, shigella flexneri, listeria monocytogenes, escherichia coli and vibrio parahaemolyticus, the growth is slow, the inhibition rate of part of the bacteria is more than 50 percent, wherein bacillus cereus, pseudomonas aeruginosa and yersinia enterocolitica can not grow at all, the escherichia coli, bacillus subtilis and vibrio parahaemolyticus are strongly inhibited when being cultured for 6h and 12h, the concentration of the non-target bacteria is far lower than that of the target bacteria, the interference of the non-target bacteria on the detection of the target bacteria is reduced, the salmonella, the staphylococcus aureus, the shigella and the listeria monocytogenes are stable in growth, and the bacterium growth speed and the bacterium growth effect are obvious.
The solution of example 3 is optimal.
According to the embodiment, the selective enrichment medium for the antagonistic antibacterial components can adsorb antibiotic residues for inhibiting the growth of microorganisms in a food sample through the resin, reduce the complexity of detection background by inhibiting the growth of non-target bacteria through the inhibitor, and enable the four target bacteria salmonella, staphylococcus aureus, shigella and listeria monocytogenes to grow rapidly through the accelerant.
The foregoing shows and describes the general principles and broad features of the present invention and advantages thereof. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (8)
1. A selective enrichment medium for antagonistic antibacterial components is characterized by comprising resin, an inhibitor and an accelerant, wherein the medium is a mixture of the resin, the inhibitor and the accelerant.
2. An antagonistic bacterium suppressing component selective enrichment medium according to claim 1, which is characterized in that: the resin is any one of Amberlyst A21 macroporous adsorption resin, DIAION HP20 macroporous adsorption resin and Amberlite IRC-84 cationic exchange resin.
3. An antagonistic bacterium suppressing component selective enrichment medium according to claim 1, which is characterized in that: the inhibitor is one or more of lithium chloride, potassium tellurite, sodium deoxycholate and ferric ammonium citrate.
4. An antagonistic bacterium suppressing component selective enrichment medium according to claim 2, which is characterized in that: the promoter is one or more of trypticase soy broth, malt extract, glucose, glutamine, sodium pyruvate, pyridoxal phosphate, vitamin B12 and sodium chloride.
5. An antagonistic bacterium suppressing component selective enrichment medium according to claim 3, which is characterized in that: the lithium chloride, potassium tellurite, sodium deoxycholate and ferric ammonium citrate are powdery particles.
6. An antagonistic bacterium suppressing component selective enrichment medium according to claim 4, which is characterized in that: the trypticase soy broth, malt extract, glucose and sodium pyruvate are powdery particles, and the lithium chloride, glutamine, pyridoxal phosphate and vitamin B12 are aqueous solutions.
7. A preparation method of a selective enrichment medium for antagonistic antibacterial components is characterized by comprising the following steps:
step 1: preparing a certain amount of distilled water, taking out lithium chloride, potassium tellurite, sodium deoxycholate and ammonium ferric citrate, pouring the lithium chloride, the potassium tellurite, the sodium deoxycholate and the ammonium ferric citrate into the distilled water step by step or at normal temperature and normal pressure, and fully stirring to dissolve the lithium chloride, the potassium tellurite, the sodium deoxycholate and the ammonium ferric citrate into the distilled water to form the inhibitor.
Step 2: preparing a certain amount of distilled water again, taking out tryptose soy broth, malt extract, glucose and sodium pyruvate, pouring the tryptose soy broth, the malt extract, the glucose and the sodium pyruvate into the distilled water gradually or a plurality of the tryptose soy broth, the malt extract, the glucose and the sodium pyruvate under normal temperature and normal pressure, fully stirring to dissolve the tryptose soy broth, the malt extract, the glucose and the sodium pyruvate into the distilled water to form an accelerating solution, filtering the lithium chloride, the glutamine, the pyridoxal phosphate and the vitamin B12 aqueous solution gradually or a plurality of the lithium chloride, the glutamine, the pyridoxal phosphate and the vitamin.
And step 3: a quantity of Amberlyst A21 macroporous adsorbent resin, DIAION HP20 macroporous adsorbent resin and Amberlite IRC-84 cationic exchange resin was used as resin.
And 4, step 4: and pouring the inhibitor and the accelerant into resin to form the selective enrichment medium for the antagonistic antibacterial components.
8. The use method of the selective enrichment medium for antagonistic antibacterial components is characterized by comprising the following steps:
step 1: salmonella, staphylococcus aureus, shigella and Listeria monocytogenes are respectively inoculated on the surface of the selective enrichment medium for antagonistic bacteriostatic components in an inclined manner, and the surface of the selective enrichment medium is subjected to gradient dilution by sterile normal saline.
Step 2: and placing the antagonistic antibacterial component selective enrichment culture medium inoculated with salmonella, staphylococcus aureus, shigella and listeria monocytogenes into a thermostat suitable for the temperature required by the growth of microorganisms for shake culture at 180 r/min.
And step 3: inoculating salmonella, staphylococcus aureus, shigella and listeria monocytogenes on resin as a contrast medium, and placing the contrast medium in a thermostat with a temperature suitable for the growth of microorganisms to carry out shake culture at 180 r/min.
And 4, step 4: the two are simultaneously shake-cultured for 24 hours; the cultures of 4h, 8h, 12h, 16h, 20h and 24h are respectively counted, and the parallel experiments are carried out for 10 times, and the thallus concentrations at different times are compared.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910933605.9A CN110592174A (en) | 2019-09-29 | 2019-09-29 | Antagonistic antibacterial component selective enrichment culture medium and preparation and use methods thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910933605.9A CN110592174A (en) | 2019-09-29 | 2019-09-29 | Antagonistic antibacterial component selective enrichment culture medium and preparation and use methods thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110592174A true CN110592174A (en) | 2019-12-20 |
Family
ID=68864948
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910933605.9A Pending CN110592174A (en) | 2019-09-29 | 2019-09-29 | Antagonistic antibacterial component selective enrichment culture medium and preparation and use methods thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110592174A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102660474A (en) * | 2012-05-05 | 2012-09-12 | 西南民族大学 | Culture medium capable of simultaneously enriching five kinds of food-borne pathogenic bacteria and preparation method for culture medium |
| CN104195086A (en) * | 2014-08-27 | 2014-12-10 | 博奥生物集团有限公司 | Composite enrichment medium for five bacteria and preparation method for composite enrichment medium |
-
2019
- 2019-09-29 CN CN201910933605.9A patent/CN110592174A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102660474A (en) * | 2012-05-05 | 2012-09-12 | 西南民族大学 | Culture medium capable of simultaneously enriching five kinds of food-borne pathogenic bacteria and preparation method for culture medium |
| CN104195086A (en) * | 2014-08-27 | 2014-12-10 | 博奥生物集团有限公司 | Composite enrichment medium for five bacteria and preparation method for composite enrichment medium |
Non-Patent Citations (3)
| Title |
|---|
| 刘园园等: "一种能同时富集沙门氏菌、金黄色葡萄球菌和单增李斯特菌的培养基", 《微生物学报》 * |
| 张巧艳等: "乳制品常见致病菌选择性共增菌技术研究", 《中国乳品工业》 * |
| 王永志等: "沙门氏菌、志贺氏菌和金黄色葡萄球菌共增菌培养基的研制", 《食品科学》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104611251A (en) | Lactic acid bacterium with wide-spectrum bacteriostatic activity and application thereof | |
| CN112501090B (en) | Bacillus licheniformis and application thereof | |
| CN115317522A (en) | Application of lactobacillus rhamnosus R7970 in preparation of pathogenic bacterium inhibition product | |
| Yang et al. | Cell age, suspending medium and metal ion influence the susceptibility of Escherichia coli O157: H7 to water-soluble maltose chitosan derivative | |
| CN104245920A (en) | Novel lactobacillus plantarum isolated from leaves of camelllia sinensis | |
| CN110592174A (en) | Antagonistic antibacterial component selective enrichment culture medium and preparation and use methods thereof | |
| CN106337027B (en) | Culture medium for isolating and detecting listeria monocytogenes | |
| CN104450506A (en) | Anaerobic blood culture medium, culture bottle and preparation method of culture bottle | |
| CN115369061A (en) | Selective helicobacter pylori culture medium and preparation method thereof | |
| CN110804646A (en) | Antibacterial activity removing method for rifampicin capsule microorganism limit control bacteria inspection | |
| US20200270670A1 (en) | Culture medium for selectively isolating bacteria of genus clostridium | |
| TW202114524A (en) | Bacillus amyloliquefaciens and use of the bacillus amyloliquefaciens and its metabolites | |
| CN109568258A (en) | A kind of amikacin sulfate injection and preparation method thereof | |
| Ronaghi et al. | A comparison of the bactericidal effectiveness of hydrochloric and acetic acid on Staphylococcus aureus in silver carp during a pH-shift protein recovery process | |
| CN103585318B (en) | Compound levofloxacin hydrochloride liposome and preparation method thereof | |
| US20160130630A1 (en) | Method for detecting campylobacter | |
| CN116687891B (en) | Application of phenyl selenium chloride | |
| Hameed | Bacteriology study of shigella species, and the effect of some ecological and chemical factors | |
| CN114836330B (en) | Preparation method and application of inhibitor of bacillus cereus | |
| CN113699085B (en) | Antagonistic bacterium ED5 and application thereof | |
| CN114343006A (en) | Compound bacteriostatic enhancer containing rosmarinic acid and application thereof | |
| JP5364308B2 (en) | Vibrio detection medium | |
| CN107739747B (en) | Method for producing reuterin at high density by co-fermentation of lactobacillus reuteri and acetobacter | |
| CN102138897B (en) | Composition preparation of lomefloxacin compound and sodium chloride and preparation method thereof | |
| CN116179639A (en) | Gram-negative bacteria selective separation culture medium and application thereof |
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 | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20191220 |
|
| WD01 | Invention patent application deemed withdrawn after publication |