CN104293712A - Deoxycholate hydrogen sulfide lactose agar medium and application thereof - Google Patents
Deoxycholate hydrogen sulfide lactose agar medium and application thereof Download PDFInfo
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- CN104293712A CN104293712A CN201410515008.1A CN201410515008A CN104293712A CN 104293712 A CN104293712 A CN 104293712A CN 201410515008 A CN201410515008 A CN 201410515008A CN 104293712 A CN104293712 A CN 104293712A
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- hydrogen sulfide
- deoxycholate
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- 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
- C12N1/20—Bacteria; Culture media therefor
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Abstract
The invention belongs to the field of microorganisms and in particular relates to a deoxycholate hydrogen sulfide lactose agar medium. The medium comprises the following components: 20.1g/L of GN enrichment fluid, 8.0g/L of lactose, 2.8g/L of sodium thiosulfate, 1.5g/L of sodium citrate, 1.5g/L of ammonium ferric citrate, 0.023g/L of neutral red and 16.5g/L of agar, wherein the pH value of the medium is 7.1-7.3; the GN enrichment fluid per liter contains 20.0g of tryptone, 2.0g of glucose, 1.5g of dipotassium hydrogen phosphate, 3g of mannitol, 5.8g of sodium citrate, 0.33g of deoxysodium cholate, 5.0g of sodium chloride and 2.0g of dipotassium hydrogen phosphate. The deoxycholate hydrogen sulfide lactose agar medium has the beneficial effects that the medium is simple in components, convenient in preparation operation and low in cost and is beneficial to popularization and application; the bacteria grow rapidly and the mutation rate is low; the medium is convenient to observe and analyze.
Description
Technical field
The present invention is microorganism field, particularly a kind of deoxycholate hydrogen sulfide lactose agar substratum and application thereof.
Background technology
The pathogenic agent of salmonellosis.Belong to enterobacteriaceae, Gram-negative enteric bacillus.Nearly 1,000 kinds (or bacterial strains) of having found.By its antigenic component, can Salmonellas be divided into first, second, third, fourth, the basic bacterium group such as penta.Wherein relevant with the human body diseases paratyphoid A bacillus mainly containing first group, the paratyphoid B bacillus of second group and bacillus typhi murium, the paratyphoid C bacillus of third group and hog cholera bacillus, the Corynebacterium diphtheriae of fourth group and bacillus enteritidis etc.Except Corynebacterium diphtheriae, paratyphoid A bacillus and paratyphoid B bacillus cause Human diseases, great majority only can cause the disease of the animals such as domestic animal, muroid and bird by order, but sometimes also can pollute the food of the mankind and cause food poisoning.Salmonellas not easily breeds in water, but can survive 2-3 week, can survive 3-4 month in refrigerator, can survive 1-2 month in the ight soil of physical environment.The suitableeest the bread worm of Salmonellas is 37 DEG C, can amount reproduction more than 20 DEG C, and therefore, low-temperature storage food is important preventive measures.
Shigella is a class gram negative bacillus, is the most common pathogenic bacteria of human bacterial's property dysentery, common name dysentery bacterium.Size is 0.5 ~ 0.7 × 2 ~ 3 μm, without gemma, without pod membrane, and atrichia.Majority has pili.Gram negative bacillus.For facultative anaerobe, can grow on ordinary culture medium, form median size, translucent smooth colony.Intestinal bacilli selective medium forms colourless bacterium colony.Biochemical reaction decomposition glucose, produces acid not aerogenesis.VP negative, not decomposing urea, do not form hydrogen sulfide, and citrate can not be utilized as carbon source.Bacillus ceylonensis A energy delayed reaction lactose (37 DEG C 3 ~ 4 days).
Substratum (Medium) is for microorganism, plant and animal tissue growth and the nutriment of artificial preparation that maintains, generally all contains carbohydrate, nitrogenous substances, inorganic salt (comprising trace element) and VITAMIN and water etc.Some substratum also contain antibiotic and pigment, for single microorganism culturing and qualification.Substratum is due to the raw material difference of preparation, and service requirements is different, and storage and custody aspect is also slightly different.General substratum, being heated, after the moisture absorption, being easily contaminated by bacterial or decomposing rotten, therefore general substratum must protection against the tide, lucifuge, the preservation of shady and cool place.Some are needed to the substratum (as tissue culture medium (TCM)) of stringent sterilization, the storage of long period, must be placed in the refrigerator of 3 ~ 6 DEG C.Because liquid nutrient medium is not easily taken care of for a long time, all change system into powder now.
Substratum is natural medium according to chemical classification, combines substratum and partly combine substratum; Be liquid nutrient medium, film solid media and dehydrated medium according to physical classification; Be selective medium, differential medium according to microorganism classification.
Summary of the invention
The present invention overcomes deficiency of the prior art, provides a kind of deoxycholate hydrogen sulfide lactose agar substratum and application thereof.
The present invention uses biological chemistry, the principle of microbiology and inorganic, organic, analytical chemistry and fluorescence technique, row filter is combined into all class nutrition compositions such as different carbon sources, nitrogenous source, inorganic salts, VITAMIN, growth stimulants, the materialization factor such as pH value, ionic strength, oxidation-reduction potential, surface tension, atmosphere surrounding of substratum is compared, investigated substratum of the present invention.
The technical solution adopted in the present invention is:
A kind of deoxycholate hydrogen sulfide lactose agar substratum, the component of described substratum comprises 20.1g/LGN enrichment liquid, 8.0g/L lactose, 2.8g/L Sulfothiorine, 1.5g/L Trisodium Citrate, 1.5g/L ferric ammonium citrate, 0.023g/L toluylene red, 16.5g/L agar, and pH value is 7.1-7.3.
Described GN enrichment liquid is containing Tryptones 20.0g, glucose 2.0g, dipotassium hydrogen phosphate 1.5g, N.F,USP MANNITOL 3g, Sodium Citrate 5.8g, sodium deoxycholate 0.33g, sodium-chlor 5.0g, dipotassium hydrogen phosphate 2.0g in often liter of GN enrichment liquid.
The invention also discloses the application of described deoxycholate hydrogen sulfide lactose agar substratum, for the selective separation of Salmonellas.
Peptone, glucose provide carbon source, nitrogenous source, VITAMIN and mineral substance; Lactose is fermentable carbohydrate; Sodium deoxycholate and Trisodium Citrate suppress gram-positive microorganism and most coliform and Bacillus proteus, but do not affect the growth of Salmonellas; Sulfothiorine and ferric ammonium citrate, for detecting the generation of hydrogen sulfide, make bacterium colony center be black; Toluylene red is pH indicator, and the bacterium colony that sugar fermentation produces acid takes on a red color, and the bacterium colony of nonfermented sugar is colourless; Agar is the peptizer of substratum.
Take this product 65.2g, heating for dissolving, in 1000ml distilled water, is waited to be chilled to 60 DEG C, impouring sterilized petri dishes.Without the need to autoclaving.
Beneficial effect of the present invention:
1, substratum prescription is simple, and preparation manipulation is convenient, and cost is low, is beneficial to and applies;
2, bacteria growing is rapid, and aberration rate is low;
3, be convenient to observe, analyze.
Embodiment
Embodiment 1:
A kind of deoxycholate hydrogen sulfide lactose agar substratum, the component of described substratum comprises 20.1g/LGN enrichment liquid, 8.0g/L lactose, 2.8g/L Sulfothiorine, 1.5g/L Trisodium Citrate, 1.5g/L ferric ammonium citrate, 0.023g/L toluylene red, 16.5g/L agar, and pH value is 7.1-7.3.
Described GN enrichment liquid is containing Tryptones 20.0g, glucose 2.0g, dipotassium hydrogen phosphate 1.5g, N.F,USP MANNITOL 3g, Sodium Citrate 5.8g, sodium deoxycholate 0.33g, sodium-chlor 5.0g, dipotassium hydrogen phosphate 2.0g in often liter of GN enrichment liquid.
Peptone, glucose provide carbon source, nitrogenous source, VITAMIN and mineral substance; Lactose is fermentable carbohydrate; Sodium deoxycholate and Trisodium Citrate suppress gram-positive microorganism and most coliform and Bacillus proteus, but do not affect the growth of Salmonellas; Sulfothiorine and ferric ammonium citrate, for detecting the generation of hydrogen sulfide, make bacterium colony center be black; Toluylene red is pH indicator, and the bacterium colony that sugar fermentation produces acid takes on a red color, and the bacterium colony of nonfermented sugar is colourless; Agar is the peptizer of substratum.
Take this product 65.2g, heating for dissolving, in 1000ml distilled water, is waited to be chilled to 60 DEG C, impouring sterilized petri dishes.Without the need to autoclaving.
18-24 hour is cultivated at 36 ± 1 DEG C
| Quality-control strains | Growing state | Further feature |
| Streptococcus aureus | - | / |
| Colon bacillus | +/- | Pink bacterium colony |
| Salmonellas | +++ | Water white transparency bacterium colony, has black center |
| Enterococcus faecalis | +/- | Water white transparency bacterium colony |
| Shigella flexneri | +++ | Water white transparency bacterium colony |
Embodiment in above-mentioned embodiment is illustrative instead of determinate; several embodiments can be listed according to institute's limited range; therefore do not departing from any change and amendment made under general plotting of the present invention, all should belong within protection domain that the present invention limits.
Claims (3)
1. a deoxycholate hydrogen sulfide lactose agar substratum, it is characterized in that the component of described substratum comprises 20.1g/LGN enrichment liquid, 8.0g/L lactose, 2.8g/L Sulfothiorine, 1.5g/L Trisodium Citrate, 1.5g/L ferric ammonium citrate, 0.023g/L toluylene red, 16.5g/L agar, pH value is 7.1-7.3.
2. deoxycholate hydrogen sulfide lactose agar substratum according to claim 1, is characterized in that described GN enrichment liquid is for containing Tryptones 20.0g, glucose 2.0g, dipotassium hydrogen phosphate 1.5g, N.F,USP MANNITOL 3g, Sodium Citrate 5.8g, sodium deoxycholate 0.33g, sodium-chlor 5.0g, dipotassium hydrogen phosphate 2.0g in often liter of GN enrichment liquid.
3. an application for deoxycholate hydrogen sulfide lactose agar substratum according to claim 1, is characterized in that the selective separation for Salmonellas.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410515008.1A CN104293712A (en) | 2014-09-29 | 2014-09-29 | Deoxycholate hydrogen sulfide lactose agar medium and application thereof |
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| CN201410515008.1A CN104293712A (en) | 2014-09-29 | 2014-09-29 | Deoxycholate hydrogen sulfide lactose agar medium and application thereof |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102492764A (en) * | 2011-12-13 | 2012-06-13 | 江门市凯林贸易有限公司 | Agar culture medium and preparation method thereof |
| CN102605039A (en) * | 2012-03-27 | 2012-07-25 | 绍兴天恒生物科技有限公司 | Salmonella culture medium containing crystal violet and production process of the culture medium |
| CN104651252A (en) * | 2013-11-25 | 2015-05-27 | 刘汉斌 | Deoxycholate hydrogen sulfide lactose agar medium and use thereof |
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2014
- 2014-09-29 CN CN201410515008.1A patent/CN104293712A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102492764A (en) * | 2011-12-13 | 2012-06-13 | 江门市凯林贸易有限公司 | Agar culture medium and preparation method thereof |
| CN102605039A (en) * | 2012-03-27 | 2012-07-25 | 绍兴天恒生物科技有限公司 | Salmonella culture medium containing crystal violet and production process of the culture medium |
| CN104651252A (en) * | 2013-11-25 | 2015-05-27 | 刘汉斌 | Deoxycholate hydrogen sulfide lactose agar medium and use thereof |
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