CN114015622A - Streptococcus agalactiae culture medium and preparation method thereof - Google Patents
Streptococcus agalactiae culture medium and preparation method thereof Download PDFInfo
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- 239000001963 growth medium Substances 0.000 title claims abstract description 36
- 241000193985 Streptococcus agalactiae Species 0.000 title claims abstract description 33
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims abstract description 22
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 claims abstract description 22
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 22
- 239000001888 Peptone Substances 0.000 claims abstract description 21
- 108010080698 Peptones Proteins 0.000 claims abstract description 21
- 235000019319 peptone Nutrition 0.000 claims abstract description 21
- 229920001817 Agar Polymers 0.000 claims abstract description 16
- 239000008272 agar Substances 0.000 claims abstract description 16
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims abstract description 11
- 229920001202 Inulin Polymers 0.000 claims abstract description 11
- 229930195725 Mannitol Natural products 0.000 claims abstract description 11
- 229910000019 calcium carbonate Inorganic materials 0.000 claims abstract description 11
- 229940041514 candida albicans extract Drugs 0.000 claims abstract description 11
- 239000008367 deionised water Substances 0.000 claims abstract description 11
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 11
- 229940029339 inulin Drugs 0.000 claims abstract description 11
- JYJIGFIDKWBXDU-MNNPPOADSA-N inulin Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)OC[C@]1(OC[C@]2(OC[C@]3(OC[C@]4(OC[C@]5(OC[C@]6(OC[C@]7(OC[C@]8(OC[C@]9(OC[C@]%10(OC[C@]%11(OC[C@]%12(OC[C@]%13(OC[C@]%14(OC[C@]%15(OC[C@]%16(OC[C@]%17(OC[C@]%18(OC[C@]%19(OC[C@]%20(OC[C@]%21(OC[C@]%22(OC[C@]%23(OC[C@]%24(OC[C@]%25(OC[C@]%26(OC[C@]%27(OC[C@]%28(OC[C@]%29(OC[C@]%30(OC[C@]%31(OC[C@]%32(OC[C@]%33(OC[C@]%34(OC[C@]%35(OC[C@]%36(O[C@@H]%37[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O%37)O)[C@H]([C@H](O)[C@@H](CO)O%36)O)[C@H]([C@H](O)[C@@H](CO)O%35)O)[C@H]([C@H](O)[C@@H](CO)O%34)O)[C@H]([C@H](O)[C@@H](CO)O%33)O)[C@H]([C@H](O)[C@@H](CO)O%32)O)[C@H]([C@H](O)[C@@H](CO)O%31)O)[C@H]([C@H](O)[C@@H](CO)O%30)O)[C@H]([C@H](O)[C@@H](CO)O%29)O)[C@H]([C@H](O)[C@@H](CO)O%28)O)[C@H]([C@H](O)[C@@H](CO)O%27)O)[C@H]([C@H](O)[C@@H](CO)O%26)O)[C@H]([C@H](O)[C@@H](CO)O%25)O)[C@H]([C@H](O)[C@@H](CO)O%24)O)[C@H]([C@H](O)[C@@H](CO)O%23)O)[C@H]([C@H](O)[C@@H](CO)O%22)O)[C@H]([C@H](O)[C@@H](CO)O%21)O)[C@H]([C@H](O)[C@@H](CO)O%20)O)[C@H]([C@H](O)[C@@H](CO)O%19)O)[C@H]([C@H](O)[C@@H](CO)O%18)O)[C@H]([C@H](O)[C@@H](CO)O%17)O)[C@H]([C@H](O)[C@@H](CO)O%16)O)[C@H]([C@H](O)[C@@H](CO)O%15)O)[C@H]([C@H](O)[C@@H](CO)O%14)O)[C@H]([C@H](O)[C@@H](CO)O%13)O)[C@H]([C@H](O)[C@@H](CO)O%12)O)[C@H]([C@H](O)[C@@H](CO)O%11)O)[C@H]([C@H](O)[C@@H](CO)O%10)O)[C@H]([C@H](O)[C@@H](CO)O9)O)[C@H]([C@H](O)[C@@H](CO)O8)O)[C@H]([C@H](O)[C@@H](CO)O7)O)[C@H]([C@H](O)[C@@H](CO)O6)O)[C@H]([C@H](O)[C@@H](CO)O5)O)[C@H]([C@H](O)[C@@H](CO)O4)O)[C@H]([C@H](O)[C@@H](CO)O3)O)[C@H]([C@H](O)[C@@H](CO)O2)O)[C@@H](O)[C@H](O)[C@@H](CO)O1 JYJIGFIDKWBXDU-MNNPPOADSA-N 0.000 claims abstract description 11
- 239000000594 mannitol Substances 0.000 claims abstract description 11
- 235000010355 mannitol Nutrition 0.000 claims abstract description 11
- 239000000843 powder Substances 0.000 claims abstract description 11
- 239000011780 sodium chloride Substances 0.000 claims abstract description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 11
- 239000012138 yeast extract Substances 0.000 claims abstract description 11
- 239000000126 substance Substances 0.000 claims abstract description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 15
- 230000001954 sterilising effect Effects 0.000 claims description 15
- 238000004659 sterilization and disinfection Methods 0.000 claims description 15
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 10
- 238000001816 cooling Methods 0.000 claims description 10
- 238000009835 boiling Methods 0.000 claims description 5
- 238000010438 heat treatment Methods 0.000 claims description 5
- 229960001855 mannitol Drugs 0.000 claims description 5
- 239000002609 medium Substances 0.000 claims description 3
- 238000000034 method Methods 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims 1
- 229960005486 vaccine Drugs 0.000 abstract description 4
- 230000001850 reproductive effect Effects 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 201000009906 Meningitis Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 206010040047 Sepsis Diseases 0.000 description 1
- 241000194017 Streptococcus Species 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 231100000225 lethality Toxicity 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 208000004396 mastitis Diseases 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 208000013223 septicemia Diseases 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
Classifications
-
- 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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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention provides a streptococcus agalactiae culture medium and a preparation method thereof, and relates to the technical field of culture media. The culture medium based on the streptococcus agalactiae and the preparation method thereof comprise the following substances in parts by weight: 3-5 parts of mannitol, 3-5 parts of inulin, 6-10 parts of yeast extract powder, 0.3-0.5 part of sodium azide, 3-5 parts of calcium carbonate, 5-7 parts of agar, 1-3 parts of peptone, 0.5-1.5 parts of sodium chloride, 15-20 parts of deionized water and 2-3 parts of pH value regulator. The culture medium can effectively facilitate the growth of the streptococcus agalactiae, and culture a high number of streptococcus agalactiae for vaccine research and use.
Description
Technical Field
The invention relates to the technical field of culture media, in particular to a streptococcus agalactiae culture medium and a preparation method thereof.
Background
Streptococcus agalactiae is a gram-positive streptococcus named because it was first isolated from cows with mastitis. Subsequently, isolated from pregnant and lying-in women and newborns in various countries, can cause early and late childhood infections, and streptococcus agalactiae is also found in various epidemic diseases of livestock and aquatic fishes, and thus has received much attention. The bacterium can cause septicemia and meningitis of animals, and has high lethality rate.
At the present stage, the use of the culture medium for the streptococcus agalactiae has certain defects, which are not beneficial to the production of the streptococcus agalactiae, the number of the cultured streptococcus agalactiae is reduced, and the research and the use of the vaccine are not facilitated, so the culture medium for the streptococcus agalactiae and the preparation method thereof are needed to solve the problem.
Disclosure of Invention
Technical problem to be solved
Aiming at the defects of the prior art, the invention provides a streptococcus agalactiae culture medium and a preparation method thereof, and solves the problem that streptococcus agalactiae cannot be effectively prepared.
(II) technical scheme
In order to achieve the purpose, the invention is realized by the following technical scheme: the streptococcus agalactiae culture medium comprises the following substances in parts by weight: 3-5 parts of mannitol, 3-5 parts of inulin, 6-10 parts of yeast extract powder, 0.3-0.5 part of sodium azide, 3-5 parts of calcium carbonate, 5-7 parts of agar, 1-3 parts of peptone, 0.5-1.5 parts of sodium chloride, 15-20 parts of deionized water and 2-3 parts of pH value regulator.
Preferably, the pH regulator is one of hydrochloric acid and sodium hydroxide.
The preparation method of the culture medium of the streptococcus agalactiae is characterized by comprising the following steps of: comprises the following preparation steps:
s1, adding mannitol, inulin, yeast extract powder, sodium azide, calcium carbonate, agar, peptone and sodium chloride into deionized water in proportion, heating and boiling until the peptone is completely dissolved, and subpackaging after the peptone is dissolved to obtain a solution;
s2, carrying out high-pressure sterilization on the solution obtained in the S1 step at the temperature of 115-121 ℃, wherein the sterilization time is 15-20 min, and cooling the solution after the sterilization is finished;
s3, shaking the sterilized solution obtained in the step S2 evenly to prevent agar from being placed at the bottom of the container to solidify, and finally obtaining a culture medium for later use.
Preferably, in the step S2, after cooling, a pH regulator is added to adjust the pH of the culture medium to 7.0 to 7.1.
(III) advantageous effects
The invention provides a culture medium for streptococcus agalactiae and a preparation method thereof. The method has the following beneficial effects:
the culture medium for the streptococcus agalactiae can effectively facilitate the growth of the streptococcus agalactiae, and can culture a high number of streptococcus agalactiae for vaccine research.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The first embodiment is as follows:
the embodiment of the invention provides a streptococcus agalactiae culture medium which comprises the following substances in parts by weight: 3 parts of mannitol, 3 parts of inulin, 6 parts of yeast extract powder, 0.3 part of sodium azide, 3 parts of calcium carbonate, 5 parts of agar, 1 part of peptone, 0.5 part of sodium chloride, 15 parts of deionized water and 2 parts of pH value regulator.
Wherein the pH regulator is hydrochloric acid or sodium hydroxide.
The preparation method of the culture medium of the streptococcus agalactiae is characterized by comprising the following steps of: comprises the following preparation steps:
s1, adding mannitol, inulin, yeast extract powder, sodium azide, calcium carbonate, agar, peptone and sodium chloride into deionized water in proportion, heating and boiling until the peptone is completely dissolved, and subpackaging after the peptone is dissolved to obtain a solution;
s2, carrying out high-pressure sterilization on the solution obtained in the S1 step at the temperature of 115-121 ℃, wherein the sterilization time is 15-20 min, and cooling the solution after the sterilization is finished;
s3, shaking the sterilized solution obtained in the step S2 evenly to prevent agar from being placed at the bottom of the container to solidify, and finally obtaining a culture medium for later use.
Wherein, in the step S2, after cooling, a pH regulator is added to adjust the pH of the culture medium to 7.0-7.1.
Example two:
the embodiment of the invention provides a streptococcus agalactiae culture medium which comprises the following substances in parts by weight: 4 parts of mannitol, 4 parts of inulin, 8 parts of yeast extract powder, 0.4 part of sodium azide, 4 parts of calcium carbonate, 6 parts of agar, 2 parts of peptone, 1 part of sodium chloride, 18 parts of deionized water and 2.5 parts of pH value regulator.
Wherein the pH regulator is hydrochloric acid or sodium hydroxide.
The preparation method of the culture medium of the streptococcus agalactiae is characterized by comprising the following steps of: comprises the following preparation steps:
s1, adding mannitol, inulin, yeast extract powder, sodium azide, calcium carbonate, agar, peptone and sodium chloride into deionized water in proportion, heating and boiling until the peptone is completely dissolved, and subpackaging after the peptone is dissolved to obtain a solution;
s2, carrying out high-pressure sterilization on the solution obtained in the S1 step at the temperature of 115-121 ℃, wherein the sterilization time is 15-20 min, and cooling the solution after the sterilization is finished;
s3, shaking the sterilized solution obtained in the step S2 evenly to prevent agar from being placed at the bottom of the container to solidify, and finally obtaining a culture medium for later use.
Wherein, in the step S2, after cooling, a pH regulator is added to adjust the pH of the culture medium to 7.0-7.1.
Example three:
the embodiment of the invention provides a streptococcus agalactiae culture medium which comprises the following substances in parts by weight: 5 parts of mannitol, 5 parts of inulin, 10 parts of yeast extract powder and 0 part of sodium azide. 5 parts of calcium carbonate, 5 parts of agar, 3 parts of peptone, 1.5 parts of sodium chloride, 20 parts of deionized water and 3 parts of pH value regulator.
Wherein the pH regulator is hydrochloric acid or sodium hydroxide.
The preparation method of the culture medium of the streptococcus agalactiae is characterized by comprising the following steps of: comprises the following preparation steps:
s1, adding mannitol, inulin, yeast extract powder, sodium azide, calcium carbonate, agar, peptone and sodium chloride into deionized water in proportion, heating and boiling until the peptone is completely dissolved, and subpackaging after the peptone is dissolved to obtain a solution;
s2, carrying out high-pressure sterilization on the solution obtained in the S1 step at the temperature of 115-121 ℃, wherein the sterilization time is 15-20 min, and cooling the solution after the sterilization is finished;
s3, shaking the sterilized solution obtained in the step S2 evenly to prevent agar from being placed at the bottom of the container to solidify, and finally obtaining a culture medium for later use.
Wherein, in the step S2, after cooling, a pH regulator is added to adjust the pH of the culture medium to 7.0-7.1.
Comparative example:
the existing culture medium A on the market is selected to be compared with the culture mediums prepared in the first embodiment, the second embodiment and the third embodiment, and the four culture mediums are tested for the reproductive capacity and the growth state (a plurality of groups of tests are carried out, the test results of each group are scored and compared, the scores are selected to be 1-10, and the average value of the scores is obtained).
As in the following table:
| reproductive capacity | Growth state | |
| Example one | 9.8 | 9.9 |
| Example two | 9.9 | 9.8 |
| EXAMPLE III | 9.8 | 9.9 |
| Medium A | 9.5 | 9.4 |
In comparison, the selected culture medium A has inferior reproductive capacity and growth state to the culture mediums prepared in the first, second and third examples, so that the culture medium for Streptococcus agalactiae of the present invention can effectively promote the growth of Streptococcus agalactiae and culture a higher amount of Streptococcus agalactiae for vaccine research.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (4)
1. The culture medium for the streptococcus agalactiae is characterized in that: the composition comprises the following substances in parts by weight: 3-5 parts of mannitol, 3-5 parts of inulin, 6-10 parts of yeast extract powder, 0.3-0.5 part of sodium azide, 3-5 parts of calcium carbonate, 5-7 parts of agar, 1-3 parts of peptone, 0.5-1.5 parts of sodium chloride, 15-20 parts of deionized water and 2-3 parts of pH value regulator.
2. The nisin-free medium according to claim 1, wherein: the pH value regulator is one of hydrochloric acid and sodium hydroxide.
3. The method for preparing a culture medium for Streptococcus agalactiae according to claim 1, wherein: comprises the following preparation steps:
s1, adding mannitol, inulin, yeast extract powder, sodium azide, calcium carbonate, agar, peptone and sodium chloride into deionized water in proportion, heating and boiling until the peptone is completely dissolved, and subpackaging after the peptone is dissolved to obtain a solution;
s2, carrying out high-pressure sterilization on the solution obtained in the S1 step at the temperature of 115-121 ℃, wherein the sterilization time is 15-20 min, and cooling the solution after the sterilization is finished;
s3, shaking the sterilized solution obtained in the step S2 evenly to prevent agar from being placed at the bottom of the container to solidify, and finally obtaining a culture medium for later use.
4. The method for preparing a culture medium for Streptococcus agalactiae according to claim 3, wherein: in the step S2, after cooling, a pH regulator is added to adjust the pH of the medium to 7.0-7.1.
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| CN202111526545.2A CN114015622A (en) | 2021-12-14 | 2021-12-14 | Streptococcus agalactiae culture medium and preparation method thereof |
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| CN202111526545.2A CN114015622A (en) | 2021-12-14 | 2021-12-14 | Streptococcus agalactiae culture medium and preparation method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1895666A (en) * | 2005-07-13 | 2007-01-17 | 崔玉东 | Cow mastitis concatenate inactivated vaccine |
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