CN104480186A - Selective chromogenic medium of bacillus cereus and preparation method thereof - Google Patents
Selective chromogenic medium of bacillus cereus and preparation method thereof Download PDFInfo
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- CN104480186A CN104480186A CN201410681386.7A CN201410681386A CN104480186A CN 104480186 A CN104480186 A CN 104480186A CN 201410681386 A CN201410681386 A CN 201410681386A CN 104480186 A CN104480186 A CN 104480186A
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Abstract
The invention provides a selective chromogenic medium of bacillus cereus. Per 1000mL of the medium comprises the following components by weight: 5-20g of heart-brain extractive, 2-6g of agar, 0.1-1g of a specific enzyme chromogenic substrate, 0.05-20g of dimethylformamide, 1-8g of glucose, 0.2-0.5g of magnesium sulfate, 0.2-0.5g of lithium chloride, 0.3-0.6g of a deoxidant, 0.2-1.0g of 3-morpholinopropane-1-sulfonic acid, 0.5-5.0g of an infectious microbe inhibitor and the balance of water, wherein the specific enzyme chromogenic substrate is a mixture of 6-chlorine-3-indolyl-beta-D-glucopyranoside and 5-bromine-4-chlorine-3-indolyl-inositol-1-potassium phosphate. By adopting the selective chromogenic medium of bacillus cereus, the defects that the manufacture cost is high and the production process is easy to pollute in the prior art can be overcome; the egg yolk or albumin is not added; the substrate is low in cost; the cost is reduced; the pollution is reduced; the selective chromogenic medium is convenient to preserve.
Description
Technical field
The present invention relates to a kind of selective coloration culture medium being applicable to bacillus cereus, and the preparation method of this selective coloration culture medium.
Background technology
Bacillus cereus (Bacillus cereus) is a kind of gram-positive bacillus, the one in Bacillus (Bacillus).Somatic cells is shaft-like, end side, becomes short or long-chain, 1.0 ~ 1.2 × 3.0 ~ 5.0 microns.Produce gemma, gemma circle or cylindricality, middle life or near middle raw, 1.0 ~ 1.5 microns, sporangiocyst is without obviously expanding.Gram-positive, without pod membrane, motion.Bacterium colony is large, and surface irregularity is flat, irregular.Wax printing fabric bacterium to external world injurious factor resistibility is strong, and distribution is wide, is typical somatic cells.
Bacillus cereus is widely distributed at occurring in nature, and some of them bacterial strain very easily contaminated food products, empty G&W etc. causes food poisoning.The data display of China's nearly 2 years food origin disease monitoring net, bacillus cereus is the another important pathogenic bacteria being only second to pathogenic colon bacillus, salmonella and Vibrio parahemolyticus, the bacterial bearing rate of rice, meat product, milk-product, veterinary antibiotics is up to 20-77%, and poisoning is to occur in the majority at school collective dining room, family, food service unit etc.Caused by the enterotoxin that the food poisoning that bacillus cereus causes produces primarily of this bacterium, be secondly that this bacterium infects, bacterium amount and toxin amount are directly proportional.Enterotoxin be divided into again thermotolerance vomiting type and thermolability diarrhea-type point, the former is mainly formed in rice based food, the type that causes vomiting after food gastroenteritis, and latter all can produce in various food, causes diarrhea-type gastroenteritis after food.
The separation and Culture of Bacillus cereus generally uses NGKG substratum or MYP substratum, these substratum contain yolk and utilize egg yellow reaction, comprise multiple steps such as separation and Culture, microscopy observation, biochemical identification, complex manufacturing technology, whole process generally needs easily pollute in production process for 4-7 days, preserve inconvenient, can not meet the reality need that microorganism quick and precisely detects.Although have color developing culture medium at present, these color developing culture mediums use the substrate of the Phospholipid hydrolase of high price, and need add albumin, cost of manufacture is high, easily pollutes in production process, preserve inconvenient.
Summary of the invention
The invention provides a kind of bacillus cereus selective coloration culture medium, and the preparation method of this selective coloration culture medium, to overcome the above-mentioned defect that prior art exists.
First the present invention relates to a kind of bacillus cereus selective coloration culture medium, and wherein every 1000mL substratum is containing, for example the component of lower weight:
Surplus is water;
Described specific enzymes chromogenic substrate is the mixture of 6-chloro-3-indyl-β-D-glucopyranoside and the bromo-4-of 5-chloro-3-indoles-inositol-1-potassiumphosphate, and both weight ratios are preferably 1:0.01-0.1, preferably 1:0.01-0.05.
Described oxygen scavenger is selected from pyruvate salt, hydrogen peroxide, thioglycolate salt, halfcystine, sodium sulphite or Iron sulfuret.
Described pyruvate salt, thioglycolate salt can be its sodium salt or sylvite.
Described miscellaneous bacteria inhibitor is trimethoprim.
The preparation method of substratum of the present invention is as follows:
1) specific enzymes chromogenic substrate is dissolved in dimethyl formamide is made into mixed solution;
2) heart-brain extract, agar, glucose, magnesium sulfate, lithium chloride, oxygen scavenger, 3-morpholino propane-1-sulfonic acid, miscellaneous bacteria inhibitor are mixed in proportion;
3) by step 1) mixed solution add step 2) in mixed solution, add water and be made into certain concentration solution;
4) cool after 120 DEG C of sterilizing 15min for subsequent use.
Bacillus cereus selective coloration culture medium of the present invention overcomes the drawbacks such as cost of manufacture in prior art is high, production process is easily polluted, and without the need to adding yolk or albumin, substrate is cheap, reduces cost reduction and pollutes convenient preservation.
Embodiment
Provide present pre-ferred embodiments below, to describe technical scheme of the present invention in detail.
Embodiment 1
Bacillus cereus selective coloration culture medium, wherein every 1000mL substratum is containing, for example the component of lower weight: the heart-brain extract 5g, the bromo-4-of agar 2g, 6-chloro-3-indyl-β-D-glucopyranoside 0.099g, 5-chloro-3-indoles-inositol-1-potassiumphosphate 0.001g, dimethyl formamide 0.05g, glucose 1-8g, magnesium sulfate 0.2g, lithium chloride 0.2g, Potassium pyruvate 0.3g, 3-morpholino propane-1-sulfonic acid 0.2g, trimethoprim 0.5-5.0g, surplus are water.
Concrete preparation method is as follows:
1) specific enzymes chromogenic substrate is dissolved in dimethyl formamide is made into mixed solution;
2) heart-brain extract, agar, glucose, magnesium sulfate, lithium chloride, oxygen scavenger, 3-morpholino propane-1-sulfonic acid, miscellaneous bacteria inhibitor are mixed in proportion;
3) by step 1) mixed solution add step 2) in mixed solution, add water and be made into certain concentration solution;
4) cool after 120 DEG C of sterilizing 15min for subsequent use.
Embodiment 2
Bacillus cereus selective coloration culture medium, wherein every 1000mL substratum is containing, for example the component of lower weight: the heart-brain extract 20g, the bromo-4-of agar 6g, 6-chloro-3-indyl-β-D-glucopyranoside 0.91g, 5-chloro-3-indoles-inositol-1-potassiumphosphate 0.09g, dimethyl formamide 20g, glucose 1-8g, magnesium sulfate 0.5g, lithium chloride 0.5g, halfcystine 0.6g, 3-morpholino propane-1-sulfonic acid 1.0g, trimethoprim 0.5-5.0g, surplus are water.
Concrete preparation method is as follows:
1) specific enzymes chromogenic substrate is dissolved in dimethyl formamide is made into mixed solution;
2) heart-brain extract, agar, glucose, magnesium sulfate, lithium chloride, oxygen scavenger, 3-morpholino propane-1-sulfonic acid, miscellaneous bacteria inhibitor are mixed in proportion;
3) by step 1) mixed solution add step 2) in mixed solution, add water and be made into certain concentration solution;
4) cool after 120 DEG C of sterilizing 15min for subsequent use.
Embodiment 3
Bacillus cereus selective coloration culture medium, wherein every 1000mL substratum is containing, for example the component of lower weight: the heart-brain extract 12g, the bromo-4-of agar 5g, 6-chloro-3-indyl-β-D-glucopyranoside 0.58g, 5-chloro-3-indoles-inositol-1-potassiumphosphate 0.02g, dimethyl formamide 0.2g, glucose 1-8g, magnesium sulfate 0.3g, lithium chloride 0.4g, Thioglycolic acid sodium salt 0.5g, 3-morpholino propane-1-sulfonic acid 0.8g, trimethoprim 0.5-5.0g, surplus are water.
Concrete preparation method is as follows:
1) specific enzymes chromogenic substrate is dissolved in dimethyl formamide is made into mixed solution;
2) heart-brain extract, agar, glucose, magnesium sulfate, lithium chloride, oxygen scavenger, 3-morpholino propane-1-sulfonic acid, miscellaneous bacteria inhibitor are mixed in proportion;
3) by step 1) mixed solution add step 2) in mixed solution, add water and be made into certain concentration solution;
4) cool after 120 DEG C of sterilizing 15min for subsequent use.
Embodiment 4
Reference culture is made respectively the standard bacteria suspension of 100cfu/mL, draw 1mL bacterium liquid and detect, cultivate 18 hours for 37 DEG C.Result is as table 1, and "+" represents positive findings, and " one " represents negative findings.
Table 1 color developing culture medium specific assay result
As can be seen from the above table, observe when pathogenic bacterium except bacillus cereus cultivate 18h on substratum, not there is specific chromogenic, and can be separated from harmless genus bacillus (such as subtilis and bacillus megaterium) and identify bacillus cereus.Visible substratum of the present invention detects bacillus cereus primary dcreening operation has excellent selectivity and specificity.
Claims (8)
1. a bacillus cereus selective coloration culture medium, is characterized in that, wherein every 1000mL substratum is containing, for example the component of lower weight:
Described specific enzymes chromogenic substrate is the mixture of 6-chloro-3-indyl-β-D-glucopyranoside and the bromo-4-of 5-chloro-3-indoles-inositol-1-potassiumphosphate.
2. bacillus cereus selective coloration culture medium as claimed in claim 1, it is characterized in that, in described specific enzymes chromogenic substrate, the weight ratio of 6-chloro-3-indyl-β-D-glucopyranoside and the bromo-4-of 5-chloro-3-indoles-inositol-1-potassiumphosphate is 1:0.01-0.1.
3. bacillus cereus selective coloration culture medium as claimed in claim 2, it is characterized in that, in described specific enzymes chromogenic substrate, the weight ratio of 6-chloro-3-indyl-β-D-glucopyranoside and the bromo-4-of 5-chloro-3-indoles-inositol-1-potassiumphosphate is 1:0.01-0.05.
4. bacillus cereus selective coloration culture medium as claimed in claim 1, it is characterized in that, described oxygen scavenger is selected from pyruvate salt, hydrogen peroxide, thioglycolate salt, halfcystine, sodium sulphite or Iron sulfuret.
5. bacillus cereus selective coloration culture medium as claimed in claim 4, it is characterized in that, described pyruvate salt is its sodium salt or sylvite.
6. bacillus cereus selective coloration culture medium as claimed in claim 4, it is characterized in that, described, thioglycolate salt is its sodium salt or sylvite.
7. bacillus cereus selective coloration culture medium as claimed in claim 1, it is characterized in that, described miscellaneous bacteria inhibitor is trimethoprim.
8. prepare a method for bacillus cereus selective coloration culture medium as described in any one of claim 1-7, it is characterized in that, comprise the steps:
1) specific enzymes chromogenic substrate is dissolved in dimethyl formamide is made into mixed solution;
2) heart-brain extract, agar, glucose, magnesium sulfate, lithium chloride, oxygen scavenger, 3-morpholino propane-1-sulfonic acid, miscellaneous bacteria inhibitor are mixed in proportion;
3) by step 1) mixed solution add step 2) in mixed solution, add water and be made into certain concentration solution;
4) cool after 120 DEG C of sterilizing 15min, namely obtain described bacillus cereus selective coloration culture medium.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106811403A (en) * | 2017-01-22 | 2017-06-09 | 贵州勤邦食品安全科学技术有限公司 | A kind of test piece of quick detection bacillus cereus and preparation method thereof, detection method |
| CN108715883A (en) * | 2018-04-24 | 2018-10-30 | 中国检验检疫科学研究院 | A kind of method of rapid screening foodborne bacterial pathogens |
| CN115315521A (en) * | 2020-03-31 | 2022-11-08 | 日水制药株式会社 | Culture medium for detecting bacillus cereus group |
Citations (1)
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| CN101215597A (en) * | 2007-12-26 | 2008-07-09 | 广东省微生物研究所 | Bacillus cereus specificity color biochemical fast detection kit and detection method |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101215597A (en) * | 2007-12-26 | 2008-07-09 | 广东省微生物研究所 | Bacillus cereus specificity color biochemical fast detection kit and detection method |
Non-Patent Citations (4)
| Title |
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| MARTINA FRICKER等: "Evaluation of standard and new chromogenic selective plating media for isolation and identification of Bacillus cereus", 《INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY》 * |
| 卢勉飞等: "蜡样芽孢杆菌显色培养基检测效果初步评价", 《食品科技》 * |
| 吴清平等: "吲哚酚显色底物的合成及其在微生物检测中的应用研究进展", 《化学通报》 * |
| 张淑红等: "应用显色培养基检测食品中蜡样芽胞杆菌的初步研究", 《现代预防医学》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106811403A (en) * | 2017-01-22 | 2017-06-09 | 贵州勤邦食品安全科学技术有限公司 | A kind of test piece of quick detection bacillus cereus and preparation method thereof, detection method |
| CN108715883A (en) * | 2018-04-24 | 2018-10-30 | 中国检验检疫科学研究院 | A kind of method of rapid screening foodborne bacterial pathogens |
| CN115315521A (en) * | 2020-03-31 | 2022-11-08 | 日水制药株式会社 | Culture medium for detecting bacillus cereus group |
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