CN104694433A - Culture medium capable of synchronously performing proliferation on a variety of pathogenic vibrio bacteria in aquatic product and preparation method thereof - Google Patents
Culture medium capable of synchronously performing proliferation on a variety of pathogenic vibrio bacteria in aquatic product and preparation method thereof Download PDFInfo
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- CN104694433A CN104694433A CN201510110499.6A CN201510110499A CN104694433A CN 104694433 A CN104694433 A CN 104694433A CN 201510110499 A CN201510110499 A CN 201510110499A CN 104694433 A CN104694433 A CN 104694433A
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- 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
- C12N1/20—Bacteria; Culture media therefor
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- 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
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- 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
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- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
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Abstract
The invention relates to a culture medium capable of synchronously performing proliferation on a variety of pathogenic vibrio bacteria in an aquatic product and a preparation method thereof. The culture medium comprises the components: 20.0 g/L of peptones and 30.0 g/L of sodium chloride. The preparation method comprises the following steps: mixing, heating and stirring the components until the components are totally dissolved; cooling to reach the room temperature; regulating a pH value to be 7.2+/-0.1; sterilizing for 15 minutes at the temperature of 121 DEG C under high pressure and cooling. According to the culture medium, composite bacteria increment of vibrio cholera, vibrio parahemolyticus, vibrio vulnificus and vibrio alginolyticus can be carried out at the same time, a complicated program of respective bacteria increment is avoided, the working efficiency is improved, and the culture medium plays an important role in quarantine inspection of pathogenic vibrio bacteria in the aquatic product.
Description
Technical field
The present invention relates to a kind of bacteria culture medium, particularly relate to a kind of can the simultaneously substratum and preparation method thereof of multiple morbid vibrio in proliferation water product.
Background technology
At present, pathogenic microbes detect is all generally carry out according to related standards method, and first standard method will increase bacterium respectively, and namely a kind of bacterium carries out increasing bacterium according to a set of enrichment medium.Pathogenic microbes detect generally will detect various pathogens simultaneously, if each pathogenic bacterium adopts a set of enrichment medium, testing can be caused loaded down with trivial details.
Table 1 GB increases bacterium method and increases comparing of bacterium method with compound
| Detection method | Detect the time limit | Sensitivity | Operability | Genome extracts |
| GB increases bacterium method | 6～24h | 10～3000 | Loaded down with trivial details (increasing bacterium 4 times) | Extract 4 |
| Compound enrichment | 6～24h | 10～230 | Simply (increase bacterium 1 time) | Extract 1 |
Summary of the invention
Based on above weak point, the invention provides a kind of can the simultaneously substratum and preparation method thereof of multiple morbid vibrio in proliferation water product; Can carry out to various pathogens such as vibrio cholerae, Vibrio parahemolyticus, Vibrio vulnificus, vibrio alginolyticus the cultivation increasing bacterium simultaneously, improve the efficiency increasing bacterium.
Object of the present invention is realized by following technology:
Can the simultaneously substratum of multiple morbid vibrio in proliferation water product, medium component: peptone 20g/L and sodium-chlor 30g/L.
The present invention also has following technical characteristic:
1, as above a kind of can the simultaneously substratum of multiple morbid vibrio in proliferation water product, its compound method is as follows:
(1) join in distilled water by peptone, sodium-chlor, heating is mixed to dissolving completely;
(2), after being cooled to room temperature, pH=7.2 ± 0.1 is adjusted;
(3) 121 DEG C of autoclaving 15min; Cooling.
2, as above a kind of can the simultaneously substratum of multiple morbid vibrio in proliferation water product, the application in the quarantine of fishery products morbid vibrio.
Beneficial effect of the present invention is embodied in: basal culture medium can carry out compound increase bacterium to vibrio cholerae, Vibrio parahemolyticus, Vibrio vulnificus, vibrio alginolyticus simultaneously, avoids the complicated procedures increasing bacterium separately and improves working efficiency; The culture increased after bacterium directly can do separation and Culture and the bioassay experiments of object bacteria, also can be directly used in PCR and detect.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited thereto.
embodiment 1
The selection of compound enrichment medium
According to the growth characteristics of vibrios in fishery products, species variation, searches compound enrichment medium, and in Bound moisture product, pathogenic bacterium subject to damage needs the features such as reparation, filters out two kinds of substratum.Two kinds of substratum Zengjing Granule vibrio cholerae, Vibrio parahemolyticus, Vibrio vulnificus, vibrio alginolyticus respectively.According to increasing bacterium situation analysis total number of bacterial colony and incubation time, determine that the substratum increasing bacterium efficiency high is optimum medium.
embodiment 2
The establishment of compound Zengjing Granule based component and compound method thereof
1, the establishment of compound Zengjing Granule based component
Medium component is: peptone 20.0g, sodium-chlor 30.0g, distilled water 1000mL.
2, the preparation method of compound enrichment medium
Concrete preparation method and cultural method as follows:
(1) join in distilled water by peptone, sodium-chlor, heating is mixed to dissolving completely;
(2), after being cooled to room temperature, pH value to 7.2 ± 0.1 is adjusted;
(3) 121 DEG C of autoclaving 15min;
(4), after substratum cooling, 18 ~ 24h is cultivated for 36 DEG C.
embodiment 3
The application of compound enrichment medium
1, with compound enrichment medium dilute respectively vibrio cholerae, Vibrio parahemolyticus, Vibrio vulnificus, vibrio alginolyticus nutrient solution to finite concentration, count.Then 36 DEG C of cultivation 24h count again, and the compound enriching effect of 4 kinds of vibrios is in table 2.Visible, increase bacterium through compound and can meet detection demand.
The compound enriching effect (unit: CFU/mL) of table 24 kinds of vibrios
| Bacterial strain | Total number of bacterial colony before increasing bacterium | Total number of bacterial colony after increasing bacterium |
| Vibrio cholerae | 220 | 2100000 |
| Vibrio parahemolyticus | 60 | 86000 |
| Vibrio vulnificus | 90 | 560000 |
| Vibrio alginolyticus | 23 | 61000 |
2, with the position reference national standard method of aseptic technique water intaking product 25g(clip sample), add after pulverizing in the aseptic triangular flask or slap type homogenizing bag filling 225mL compound enrichment medium, in compound enrichment medium, be vaccinated with vibrio cholerae, Vibrio parahemolyticus, Vibrio vulnificus, vibrio alginolyticus 4 kinds of positive bacterias.Rock 3 ~ 5min or bounce 1min, cultivating 4 ~ 6h by after triangular flask or homogenizing bag sealing at 36 DEG C.From compound enrichment medium, take out 10 mL join in the test tube filling 10mL compound increasing bacterium base, after mixing, cultivate 18 ~ 24h at 36 DEG C.From complex medium, extract the nucleic acid of vibrios, carry out the PCR detection of vibrio cholerae, Vibrio parahemolyticus, Vibrio vulnificus, vibrio alginolyticus, the positive bacteria of result inoculation all detects.
Claims (3)
1. the energy simultaneously substratum of multiple morbid vibrio in proliferation water product, it is characterized in that, medium component is: peptone 20g/L and sodium-chlor 30g/L.
2. according to claim 1 a kind of can the simultaneously substratum of multiple morbid vibrio in proliferation water product, it is characterized in that, compound method is as follows:
(1) join in distilled water by peptone, sodium-chlor, heating is mixed to dissolving completely;
(2), after being cooled to room temperature, pH=7.2 ± 0.1 is adjusted;
(3) 121 DEG C of autoclaving 15min; Cooling.
3. according to claim 1 a kind of can the simultaneously substratum of multiple morbid vibrio in proliferation water product, it is characterized in that, the application in the quarantine of fishery products morbid vibrio.
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| CN201510110499.6A CN104694433A (en) | 2015-03-13 | 2015-03-13 | Culture medium capable of synchronously performing proliferation on a variety of pathogenic vibrio bacteria in aquatic product and preparation method thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105087749A (en) * | 2015-08-06 | 2015-11-25 | 温州医科大学附属第一医院 | Preenrichment liquid culture medium for vibrio vulnificus clinical detection and application of preenrichment liquid culture medium |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101412977A (en) * | 2008-11-25 | 2009-04-22 | 华南理工大学 | Culture medium for composite enrichment of salmonella, Vibrio parahaemolyticus and Vibrio cholerae, and preparation thereof |
| CN103923857A (en) * | 2014-04-21 | 2014-07-16 | 山东大学 | Vibrio FC509 strain and culturing method and application thereof |
| CN104195213A (en) * | 2014-09-02 | 2014-12-10 | 青岛永通电梯工程有限公司 | Vibrio chromogenic culture medium |
| CN104293880A (en) * | 2014-10-28 | 2015-01-21 | 天津大学 | Seawater selective chromogenic culture media for VP (Vibrio Parahaemolyticus) and authenticating and counting method thereof |
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2015
- 2015-03-13 CN CN201510110499.6A patent/CN104694433A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101412977A (en) * | 2008-11-25 | 2009-04-22 | 华南理工大学 | Culture medium for composite enrichment of salmonella, Vibrio parahaemolyticus and Vibrio cholerae, and preparation thereof |
| CN103923857A (en) * | 2014-04-21 | 2014-07-16 | 山东大学 | Vibrio FC509 strain and culturing method and application thereof |
| CN104195213A (en) * | 2014-09-02 | 2014-12-10 | 青岛永通电梯工程有限公司 | Vibrio chromogenic culture medium |
| CN104293880A (en) * | 2014-10-28 | 2015-01-21 | 天津大学 | Seawater selective chromogenic culture media for VP (Vibrio Parahaemolyticus) and authenticating and counting method thereof |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105087749A (en) * | 2015-08-06 | 2015-11-25 | 温州医科大学附属第一医院 | Preenrichment liquid culture medium for vibrio vulnificus clinical detection and application of preenrichment liquid culture medium |
| CN105087749B (en) * | 2015-08-06 | 2018-01-19 | 温州医科大学附属第一医院 | For the preceding enrichment liquid body culture medium of Vibrio vulnificus clinical detection and application |
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