CN106282075A - A kind of fluid medium for cultivating haemophilus parasuis - Google Patents
A kind of fluid medium for cultivating haemophilus parasuis Download PDFInfo
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- CN106282075A CN106282075A CN201610970214.0A CN201610970214A CN106282075A CN 106282075 A CN106282075 A CN 106282075A CN 201610970214 A CN201610970214 A CN 201610970214A CN 106282075 A CN106282075 A CN 106282075A
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- 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 present invention provides the fluid medium that a kind of haemophilus parasuis is cultivated, and it includes following component: tryptone, soy peptone, sodium chloride, dipotassium hydrogen phosphate, glucose, yeast powder, Ox blood serum and cozymase, its pH value is 7.2～7.4.The culture medium of the present invention is by supplementing the nutritional labeling that various Institute of Micro-biology need, and is effectively controlled the growth of the pH value in culture medium, beneficially haemophilus parasuis by sodium hydroxide, and viable count is all not less than 109CFU/ml, and haemophilus parasuis bacterium solution is prepared inactivated vaccine, its safety and immune efficacy are the best.
Description
Technical field
The invention belongs to cause of disease culture technique field, be specifically related to a kind of liquid culture for cultivating haemophilus parasuis
Base.
Background technology
Pig Haemophilus parasuis, also known as pig leather La Sishi sick (Disease), it is by haemophilus parasuis
The infectious disease being characterized with polyserositis, arthritis and meningitis caused, is the world of serious harm in recent years pig industry
One of significant bacterial sexually transmitted disease of development.At present, it has already been proven that haemophilus parasuis China overwhelming majority pig farm exist and
Popular, mainly affect the piglet of 5～8 week old, sickness rate is generally l0%～15%, and time serious, mortality rate is up to more than 50%,
Huge economic loss is caused to China's pig industry.But there is a lot of problem, such as in the cultivation currently for haemophilus parasuis
Culture medium cost is high, nutritional requirement is high, pH value requires strict, and culture density is low.This all research to this vaccine causes seriously
Impact, therefore develops the high-density culture medium of a kind of haemophilus parasuis, and tool is of great significance.
Summary of the invention
The goal of the invention of the present invention is for deficiency of the prior art, it is provided that a kind of high-effect, secondary pig of low cost
The preparation of haemophilus fluid medium, thus make up the deficiencies in the prior art.
The fluid medium of the present invention, it includes following component: tryptone, soy peptone, sodium chloride, phosphoric acid
Hydrogen dipotassium, glucose, yeast powder, Ox blood serum and cozymase, its pH value is 7.2～7.4.
Its concentration of component is tryptone 18～25.0g/L, soy peptone 3.0～5.0g/L, sodium chloride 5.0g/L, phosphorus
Acid hydrogen dipotassium 3.0～4.0g/L, glucose 2.0～3.0g/L, yeast powder 1.0～5.0g/L, inactivation Ox blood serum 30ml/L～
100ml/L, cozymase 1.0～30g/L, pH value is adjusted to 7.2～7.4.
As preferably, its concentration of component is tryptone 22.0g/L, soy peptone 4.0g/L, sodium chloride 5.0g/L, phosphorus
Acid hydrogen dipotassium 3.0g/L, glucose 2.5g/L, yeast powder 3.0g/L, inactivation Ox blood serum 50ml/L, cozymase 0.1g/L, pH value is adjusted
To 7.2～7.4.
The preparation method of above-mentioned culture medium is as follows:
Weigh tryptone, soybean protein, sodium chloride, dipotassium hydrogen phosphate, glucose, yeast powder respectively, add deionization
Water, fully shakes up dissolving, tune pH value to 7.2～7.4,121 DEG C of high pressure steam sterilization 15min after constant volume；After cooling, want by aseptic
Ask addition inactivation Ox blood serum and the 1%NAD cozymase of filtration sterilization.
The culture medium of the present invention is by supplementing the nutritional labeling that various Institute of Micro-biology need, and is effectively controlled by sodium hydroxide
The growth of the pH value in culture medium processed, beneficially haemophilus parasuis, viable count is all not less than 109CFU/ml, and by pair pig addicted to
Blood bacillus bacterium solution prepares inactivated vaccine, and its safety and immune efficacy are the best.
Detailed description of the invention
Below in conjunction with embodiment, the present invention is described in detail.
Embodiment 1 prepares culture medium of the present invention according to different formulations
1 culture medium preparation is by the culture medium of different culture medium prescription 6 kinds of heterogeneities of preparation.Concrete formula is shown in Table 1.
Table 1 fluid medium preparation method
The screening of embodiment 2 different formulations fluid medium:
By haemophilus parasuis YBH05 strain, (haemophilus parasuis delivers Chaoyang District, Beijing City on November 23rd, 2011
No. 3 China Committee for Culture Collection of Microorganisms of Institute of Microorganism, Academia Sinica common micro-organismss of North Star West Road 1 institute
Center preservation, preserving number is: CGMCC No.5480) bacterium solution, respectively by the 5% above-mentioned 6 kinds of fluid mediums of inoculation, put 37 DEG C,
150r/min shaken cultivation, incubation time is 24 hours, and in cultivating, different time is separately sampled carries out count plate.Difference is joined
The screening test result of side's fluid medium shows, preferable according to the fluid medium culture effect of formula three, formula four preparation,
Within 14～16 hours, viable count is all not less than 109CCU/ml, formula five and formula six are because of auxiliary liquid compositions such as the serum in culture medium
Addition is higher, makes culture medium nutrition the abundantest, causes YBH05 strain growth too fast, and decline is also very fast, and the auxiliary liquid such as serum becomes
Divide addition high, too increase production cost；It is relatively low that formula one and formula two cultivate bacterium solution density.Consider, finally select
Culture effect verified by the fluid medium of formula three preparation.The results are shown in Table 2.
Different time count plate result cultivated by table 2:6 kind fluid medium
Embodiment 3: culture medium of the present invention compares with pancreas peptone soybean broth fluid medium culture effect
YBH05 strain freeze-dried semen is recovered after purification, inoculates pancreas peptone soybean broth fluid medium by 3-5% respectively
With the fluid medium of the formula three of the present invention, putting 37 DEG C respectively, 150r/min shaken cultivation 24 hours, in cultivating different time
Separately sampled carry out count plate.Result of the test shows, applies culture medium of the present invention, the viable bacteria amount of different incubation times to be above
Pancreas peptone soybean broth fluid medium.The viable bacteria amount of different incubation times the results are shown in Table 3.
Table 3: the Selection experiment result of culture medium
Embodiment 4:
3 batches of YBH05 strain bacterium solution of culture medium fermentation culture of the checking application inventive formulation three of culture medium of the present invention, respectively
Carry out count plate, to verify the feasibility of culture process.The result shows, the bacterium solution viable bacteria amount of YBH05 strain be 2.5～
3.0×109CFU/ml, the culture effect of culture medium of the present invention is truly feasible.The results are shown in Table 4.
Table 4 the result of the present invention
Other haemophilus parasuis is used to also indicate that the culture medium of the present invention also has very well as the culture effect of seed
Effect.
Be can be seen that culture medium of the present invention incubation time in incubation is short by above-mentioned result of the test, growth is fast, viable bacteria
Amount height, the large-scale production being especially suitable for haemophilus parasuis is cultivated.
Claims (6)
1. a fluid medium, it is characterised in that described fluid medium its include following component: tryptone,
Soy peptone, sodium chloride, dipotassium hydrogen phosphate, glucose, yeast powder, Ox blood serum and cozymase, its pH value is 7.2～7.4.
2. fluid medium as claimed in claim 1, it is characterised in that described fluid medium, its concentration of component is as follows:
Tryptone 18～25.0g/L, soy peptone 3.0～5.0g/L, sodium chloride 5.0g/L, dipotassium hydrogen phosphate 3.0～4.0g/L,
Glucose 2.0～3.0g/L, yeast powder 1.0～5.0g/L, inactivation Ox blood serum 30ml/L～100ml/L, cozymase 1.0～30g/
L, pH value is adjusted to 7.2～7.4.
3. fluid medium as claimed in claim 1 or 2, it is characterised in that described fluid medium, its concentration of component is such as
Under: tryptone 22.0g/L, soy peptone 4.0g/L, sodium chloride 5.0g/L, dipotassium hydrogen phosphate 3.0g/L, glucose 2.5g/
L, yeast powder 3.0g/L, inactivation Ox blood serum 50ml/L, cozymase 0.1g/L, pH value is adjusted to 7.2～7.4.
4. the preparation method of the fluid medium described in any one of claim 1-3, it is characterised in that described method include as
Under step:
Weigh tryptone, soy peptone, sodium chloride, dipotassium hydrogen phosphate, glucose, yeast powder respectively, add deionized water
Fully shake up dissolving, tune pH value to 7.2～7.4,121 DEG C of high pressure steam sterilization 15min.After cooling, add by sterility requirements
50ml inactivates Ox blood serum, the NAD of 10ml filtration sterilization.
5. preparation method as claimed in claim 4, it is characterised in that after culture medium high pressure, after cooling, adds inactivation Ox blood serum
And NAD.
6. the application in cultivating haemophilus parasuis of the fluid medium described in any one of claim 1-3.
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| CN201610970214.0A CN106282075A (en) | 2016-11-02 | 2016-11-02 | A kind of fluid medium for cultivating haemophilus parasuis |
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| CN201610970214.0A CN106282075A (en) | 2016-11-02 | 2016-11-02 | A kind of fluid medium for cultivating haemophilus parasuis |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107177531A (en) * | 2017-06-15 | 2017-09-19 | 广东海大畜牧兽医研究院有限公司 | A kind of growth accelerator for improving haemophilus parasuis in vitro culture |
| CN108977392A (en) * | 2018-08-14 | 2018-12-11 | 沈阳农业大学 | A kind of haemophilus parasuis proliferated culture medium and preparation method thereof |
| CN109207397A (en) * | 2018-09-18 | 2019-01-15 | 山东省滨州畜牧兽医研究院 | A kind of haemophilus parasuis culture medium and high density fermentation culture method |
| CN112831428A (en) * | 2019-11-25 | 2021-05-25 | 北京信得威特科技有限公司 | Haemophilus parasuis culture medium |
| CN113481120A (en) * | 2021-06-29 | 2021-10-08 | 广州知易生物科技有限公司 | Culture medium, preparation method thereof and method for culturing bacteroides fragilis by using culture medium |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107177531A (en) * | 2017-06-15 | 2017-09-19 | 广东海大畜牧兽医研究院有限公司 | A kind of growth accelerator for improving haemophilus parasuis in vitro culture |
| CN107177531B (en) * | 2017-06-15 | 2018-07-06 | 广东海大畜牧兽医研究院有限公司 | A kind of growth accelerator for improving haemophilus parasuis in vitro culture |
| CN108977392A (en) * | 2018-08-14 | 2018-12-11 | 沈阳农业大学 | A kind of haemophilus parasuis proliferated culture medium and preparation method thereof |
| CN108977392B (en) * | 2018-08-14 | 2021-08-06 | 沈阳农业大学 | Haemophilus parasuis proliferation medium and preparation method thereof |
| CN109207397A (en) * | 2018-09-18 | 2019-01-15 | 山东省滨州畜牧兽医研究院 | A kind of haemophilus parasuis culture medium and high density fermentation culture method |
| CN109207397B (en) * | 2018-09-18 | 2020-07-28 | 山东省滨州畜牧兽医研究院 | Haemophilus parasuis culture medium and high-density fermentation culture method |
| CN112831428A (en) * | 2019-11-25 | 2021-05-25 | 北京信得威特科技有限公司 | Haemophilus parasuis culture medium |
| CN113481120A (en) * | 2021-06-29 | 2021-10-08 | 广州知易生物科技有限公司 | Culture medium, preparation method thereof and method for culturing bacteroides fragilis by using culture medium |
| CN113481120B (en) * | 2021-06-29 | 2023-03-21 | 广州知易生物科技有限公司 | Culture medium, preparation method thereof and method for culturing bacteroides fragilis by using culture medium |
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