CN102492764A - Agar culture medium and preparation method thereof - Google Patents
Agar culture medium and preparation method thereof Download PDFInfo
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- CN102492764A CN102492764A CN2011104140765A CN201110414076A CN102492764A CN 102492764 A CN102492764 A CN 102492764A CN 2011104140765 A CN2011104140765 A CN 2011104140765A CN 201110414076 A CN201110414076 A CN 201110414076A CN 102492764 A CN102492764 A CN 102492764A
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Abstract
The invention discloses an agar culture medium. Each 1,000 milliliters of nutrient solution comprises the following components by mass: 3 to 7 grams of peptone, 8 to 12 grams of lactose, 3 to 7 grams of beef extract powder, 3 to 7 grams of No.3 cholate, 0.023 to 0.027 gram of neutral red, 13 to 17 grams of agar, 7.5 to 9.5 grams of sodium citrate, 1.3 to 1.7 grams of ferric citrate, 0.0003 to 0.00036 gram of brilliant green and 8.2 to 8.8 grams of sodium thiosulfate. The preparation method comprises the following steps of: 1) dissolving 5 grams of peptone, 10 grams of lactose, 5 grams of beef extract powder, 5 grams of No.3 cholate, 15 grams of agar, 8 grams of sodium citrate, 1.5 grams of ferric citrate and 8.5 grams of sodium thiosulfate in 1,000 milliliters of nutrient solution; 2) heating; 3) regulating the pH value to be between 7.0+/-0.1, and adding 0.025 gram of neutral red and 0.00033 gram of brilliant green; 4) sterilizing; and 5) refrigerating.
Description
Technical field
The present invention relates to a kind of agar culture technique field, nutrient agar of especially a kind of separation and Culture that is used for Salmonellas and Shigellae and preparation method thereof.
Background technology
The situation that this product uses on the market is not obvious as the evil mind in the assay bacterium colony of hydrogen sulfide male bacterium, and is very fuzzy, even evil mind is arranged, but the also not enough bacterium colony of its black area 1/3rd, cause the omission phenomenon easily.
Summary of the invention
The object of the invention just provides a kind of nutrient agar and preparation method thereof, increases the nutritive substance of product, improves the recall rate of object bacteria, solves the phenomenon of omission.
In order to reach above-mentioned purpose of design, the technical scheme that the present invention adopts is following:
A kind of nutrient agar, its moity and total mass number are: in every 1000ml nutritive medium: peptone 3g-7g, lactose 8g-12g, beef extract powder 3g-7g, no. 3 bile salt 3g-7g, toluylene red 0.023g-0.027g, agar 13g-17g, Trisodium Citrate 7.5g-9.5g, ironic citrate 1.3g-1.7g, brilliant green 0.0003g-0.00036g, Sulfothiorine 8.2g-8.8g.
Preferably, said its moity and total mass number are: in every 1000ml nutritive medium: peptone 3g-7g, lactose 10g, beef extract powder 5g, no. 3 bile salt 5g, toluylene red 0.05g, agar 15g, Trisodium Citrate 8g, ironic citrate 15g, brilliant green 0.00033g, Sulfothiorine 8.5g.
Preferably, the staple of said nutritive medium is amino acid, VITAMINs, growth factor.
A kind of making method of nutrient agar may further comprise the steps:
1) peptone 5g, lactose 10g, beef extract powder 5g, no. 3 bile salt 5g, agar 15g, Trisodium Citrate 8g, ironic citrate 1.5g, Sulfothiorine 8.5g are dissolved in the 1000ml nutritive medium;
2) behind the mixing, heated and boiled is fully dissolved;
3) regulate pH to 7.0 ± 0.1.Add toluylene red 0.025g and brilliant green 0.00033g;
4) boiling sterilization 1min-2min is cooled to about 50 ℃, is poured in the disposable sterilized plastics plate;
5) behind the steriling test in 2 ℃ of-8 ℃ of environment refrigeration subsequent use.
Preferably, the method for said adjustment pH to 7.0 ± 0.1 is to regulate pH to 7.0 ± 0.1 with 1N-10N sodium hydroxide or 1N-10N hydrochloric acid soln.
Its principle is: beef extract powder and peptone provide required carbon source of bacterial growth and nitrogenous source as base nutrients matter in substratum; The growth of no. 3 bile salt and brilliant green inhibition gram-positive microorganism and most of coliform; When hydrogen sulfide male bacterium grew on this substratum, the sulfur reduction in the Sulfothiorine became S
2-, S
2-Be combined into the FeS black precipitate with the iron ion in the ironic citrate; Adhere to bacterium colony and make the bacterium colony black in color; Lactose is as fermentable glucide, and the coliform bacterioid utilizes lactose to produce toluylene red indicator that acid the makes substratum look that reddens, thereby makes bacterium colony manifest redness; In addition, make cholate be deposited in periphery of bacterial colonies formation cholate precipitation ring owing to produce acid.
The beneficial effect of nutrient agar of the present invention and preparation method thereof is: the area of hydrogen sulfide male bacterium black reaches 4/5ths of bacterium itself, makes object bacteria be more prone to observe, and has solved the problem of omission; The bacterium colony size is better than similar products at home and abroad; The time ratio like product of sample sentence read result is shorter; Salmonellas bacterium colony evil mind is more obvious than similar products at home and abroad.
Embodiment
Embodiment 1,
A kind of nutrient agar, its moity and total mass number are: in every 1000ml nutritive medium: peptone 3g, lactose 8g, beef extract powder 3g, no. 3 bile salt 7g, toluylene red 0.027g, agar 17g, Trisodium Citrate 9.5g, ironic citrate 1.7g, brilliant green 0.00036g, Sulfothiorine 8.8g.
Embodiment 2,
A kind of nutrient agar, its moity and total mass number are: in every 1000ml nutritive medium: peptone 7g, lactose 12g, beef extract powder 3g, no. 3 bile salt 3g, toluylene red 0.023g, agar 1, Trisodium Citrate 7.5g, ironic citrate 1.3g, brilliant green 0.0003g, Sulfothiorine 8.2g.
Embodiment 3,
A kind of nutrient agar, its moity and total mass number are: in every 1000ml nutritive medium: peptone 3g-7g, lactose 10g, beef extract powder 5g, no. 3 bile salt 5g, toluylene red 0.05g, agar 15g, Trisodium Citrate 8g, ironic citrate 15g, brilliant green 0.00033g, Sulfothiorine 8.5g.
Among above-mentioned three embodiment, the staple of said nutritive medium is amino acid, VITAMINs, growth factor.
Embodiment 4,
A kind of making method of nutrient agar may further comprise the steps:
1) peptone 3g, lactose 8g, beef extract powder 3g, no. 3 bile salt 7g, agar 17g, Trisodium Citrate 9.5g, ironic citrate 1.7g, Sulfothiorine 8.8g are dissolved in the 1000ml nutritive medium;
2) behind the mixing, heated and boiled is fully dissolved;
3) regulate pH to 7.0 ± 0.1.Add toluylene red 0.027g and brilliant green 0.00036g;
4) boiling sterilization 1min-2min is cooled to about 50 ℃, is poured in the disposable sterilized plastics plate;
5) behind the steriling test in 2 ℃ of-8 ℃ of environment refrigeration subsequent use.
Embodiment 5,
A kind of making method of nutrient agar may further comprise the steps:
1) peptone 7g, lactose 12g, beef extract powder 3g, no. 3 bile salt 3g, agar 15g, Trisodium Citrate 7.5g, ironic citrate 1.3g, Sulfothiorine 8.2g are dissolved in the 1000ml nutritive medium;
2) behind the mixing, heated and boiled is fully dissolved;
3) regulate pH to 7.0 ± 0.1.Add toluylene red 0.023g and brilliant green 0.0003g;
4) boiling sterilization 1min-2min is cooled to about 50 ℃, is poured in the disposable sterilized plastics plate;
5) behind the steriling test in 2 ℃ of-8 ℃ of environment refrigeration subsequent use.
Embodiment 6,
A kind of making method of nutrient agar may further comprise the steps:
1) peptone 5g, lactose 10g, beef extract powder 5g, no. 3 bile salt 5g, agar 15g, Trisodium Citrate 8g, ironic citrate 1.5g, Sulfothiorine 8.5g are dissolved in the 1000ml nutritive medium;
2) behind the mixing, heated and boiled is fully dissolved;
3) regulate pH to 7.0 ± 0.1.Add toluylene red 0.025g and brilliant green 0.00033g;
4) boiling sterilization 1min-2min is cooled to about 50 ℃, is poured in the disposable sterilized plastics plate;
5) behind the steriling test in 2 ℃ of-8 ℃ of environment refrigeration subsequent use.
In the foregoing description, the method for said adjustment pH to 7.0 ± 0.1 is to regulate pH to 7.0 ± 0.1 with 1N-10N sodium hydroxide or 1N-10N hydrochloric acid soln.
This embodiment the preferred embodiments of the present invention can not limit the present invention, and concrete each item rights protection scope is defined by the claims.
Claims (5)
1. nutrient agar, it is characterized in that: its moity and total mass number are: in every 1000ml nutritive medium: peptone 3g-7g, lactose 8g-12g, beef extract powder 3g-7g, no. 3 bile salt 3g-7g, toluylene red 0.023g-0.027g, agar 13g-17g, Trisodium Citrate 7.5g-9.5g, ironic citrate 1.3g-1.7g, brilliant green 0.0003g-0.00036g, Sulfothiorine 8.2g-8.8g.
2. nutrient agar according to claim 1 is characterized in that: said its moity and total mass number are: in every 1000ml nutritive medium: peptone 5g, lactose 10g, beef extract powder 5g, no. 3 bile salt 5g, toluylene red 0.05g, agar 15g, Trisodium Citrate 8g, ironic citrate 15g, brilliant green 0.00033g, Sulfothiorine 8.5g.
3. nutrient agar according to claim 1 and 2 is characterized in that: the staple of said nutritive medium is amino acid, VITAMINs, growth factor.
4. the making method of a nutrient agar is characterized in that: may further comprise the steps:
1) peptone 5g, lactose 10g, beef extract powder 5g, no. 3 bile salt 5g, agar 15g, Trisodium Citrate 8g, ironic citrate 1.5g, Sulfothiorine 8.5g are dissolved in the 1000ml nutritive medium;
2) behind the mixing, heated and boiled is fully dissolved;
3) regulate pH to 7.0 ± 0.1, add toluylene red 0.025g and brilliant green 0.00033g;
4) boiling sterilization 1min-2min is cooled to about 50 ℃, is poured in the disposable sterilized plastics plate;
5) behind the steriling test in 2 ℃ of-8 ℃ of environment refrigeration subsequent use.
5. the making method of nutrient agar according to claim 4 is characterized in that: the method for said adjustment pH to 7.0 ± 0.1 is, with 1N-10N sodium hydroxide or 1N-10N hydrochloric acid soln adjusting pH to 7.0 ± 0.1.
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| CN2011104140765A CN102492764A (en) | 2011-12-13 | 2011-12-13 | Agar culture medium and preparation method thereof |
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| CN2011104140765A CN102492764A (en) | 2011-12-13 | 2011-12-13 | Agar culture medium and preparation method thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104293712A (en) * | 2014-09-29 | 2015-01-21 | 青岛康合伟业商贸有限公司 | Deoxycholate hydrogen sulfide lactose agar medium and application thereof |
-
2011
- 2011-12-13 CN CN2011104140765A patent/CN102492764A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 中华人民共和国***医政司: "《全国临床检验操作规程（第二版）》", 31 December 1997, 东南大学出版社 * |
| 刘坚真 等: "国家标准测定食品细菌总数培养基的改进研究", 《微生物学通报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104293712A (en) * | 2014-09-29 | 2015-01-21 | 青岛康合伟业商贸有限公司 | Deoxycholate hydrogen sulfide lactose agar medium and application thereof |
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Application publication date: 20120613 |