CN104046582A - Medium rich in Pseudomonas otitidis - Google Patents
Medium rich in Pseudomonas otitidis Download PDFInfo
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- CN104046582A CN104046582A CN201410245469.1A CN201410245469A CN104046582A CN 104046582 A CN104046582 A CN 104046582A CN 201410245469 A CN201410245469 A CN 201410245469A CN 104046582 A CN104046582 A CN 104046582A
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- pseudomonas
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- otitis
- peptone
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Abstract
The invention discloses a medium rich in Pseudomonas otitidis. The medium comprises a basic medium, glucose, sodium bicarbonate, peptone, ammonium nitrate, a yeast extract and vitamin D; and 1.0-5.0g of glucose, 0.1-0.5g of sodium bicarbonate, 1.0-5.0g of peptone, 0.1-0.5g of ammonium nitrate, 0.05-0.1g of the yeast extract and 0.05-0.1g of vitamin D are added to each 1000mL of the basic medium. The medium disclosed in the invention is used to culture rich Pseudomonas otitidis, and has the advantages of fast culturing speed and low cost, makes up the disadvantages of too slow Pseudomonas otitidis growth period, improves the growth rate, and shortens the time reaching largest yield to 9-11h.
Description
Technical field
The present invention relates to a kind of substratum of enrichment of bacterial, be specifically related to the substratum of a kind of enrichment otitis pseudomonas.
Background technology
Otitis pseudomonas (Pseudomonas otitidis.) is the bacterial strain of the novel Rhodopseudomonas that identified in 2006, this bacterial strain is to separate and obtain the people's ear clinical sample from infecting for the first time, and a kind of virulence is lower but bacillus that resistance is strong.This bacterial strain is a kind of conditioned pathogen, only under certain specified conditions, could cause a disease.And otitis pseudomonas has the symbolic characteristic of potential industrial applicability, this bacterial strain can be used for water body purification, there is the function of aerobic denitrification, nutritional needs is simple, therefore, application prospect is very wide, and some pollution substances in otitis pseudomonas degradable waste water, therefore have again very high research and using value.The promotion and application of otitis pseudomonas, key is to need suitable culture medium, because otitis pseudomonas is excessively slow vegetative period, the substratum of the enrichment otitis pseudomonas adopting at present, its average enrichment time is 28～35d, the cycle is all longer.
Summary of the invention
In order to solve defect and the deficiency of prior art existence, the invention provides the substratum of a kind of enrichment otitis pseudomonas, with culture medium culturing enrichment otitis pseudomonas of the present invention, cultivation speed is fast, and cost is low.Made up the weak point that it's vegetative period pastes the aspect such as slow otitis pseudomonas, improved growth velocity, make to reach maximum growth amount time shorten 9～11 hours.
The technical solution adopted in the present invention is: the substratum of a kind of enrichment otitis pseudomonas, comprise basic medium, glucose, sodium bicarbonate, peptone, ammonium nitrate, yeast extract paste and vitamins D, every 1000mL basic medium adds glucose 1.0～5.0g, sodium bicarbonate 0.1-0.5g, peptone 1.0～5.0g, ammonium nitrate 0.1-0.5g, yeast extract paste 0.05～0.1g and vitamins D 0.05～0.1g.
Preferably, every 1000mL basic medium adds glucose 3.0g, sodium bicarbonate 0.3g, peptone 3.0g, ammonium nitrate 0.3g, yeast extract paste 0.07g and vitamins D 0.07g.
Basic medium is the conventional substratum of pseudomonas.Can select in prior art through conventional, and published pseudomonas substratum.
Utilize the method for above-mentioned substratum enrichment culture otitis pseudomonas: by pH regulator to 7.0～7.2 of the substratum of above-mentioned enrichment otitis pseudomonas, 121 DEG C of sterilizing 30min, otitis pseudomonas is inoculated in by 1% inoculum size in the substratum of enrichment otitis pseudomonas, vibration mixes, and in temperature is the constant incubator of 30 DEG C, cultivates.
The impact of the increment of glucose on otitis pseudomonas increases along with the increase of the concentration of glucose, after the lucifuge of 20h is cultivated, in the time that the consumption of glucose is 3.0g/L (every 1000mL basic medium adds 3.0g), its thalline reproduction speed is very fast, bacterium liquid OD
600reach 1.50, in the time that the consumption of glucose is less than 3.0g/L, bacterium liquid OD
600be less than 1.50.Therefore, the consumption of glucose is advisable with 3.0g/L.
The impact of the increment of sodium bicarbonate on otitis pseudomonas increases along with the increase of the concentration of sodium bicarbonate, after the lucifuge of 20h is cultivated, in the time that the consumption of sodium bicarbonate is 0.3g/L (every 1000mL basic medium adds 0.3g), its thalline reproduction speed is very fast, bacterium liquid OD
600reach 1.50, in the time that the consumption of sodium bicarbonate is less than 0.3g/L, bacterium liquid OD
600be less than 1.50.Therefore, the consumption of sodium bicarbonate is advisable with 0.3g/L.
The impact of the increment of peptone on otitis pseudomonas, increase along with the increase of the concentration of peptone, after the lucifuge of 20h is cultivated, in the time that the consumption of peptone is 3.0g/L (every 1000mL basic medium adds 3.0g), its thalline reproduction speed is very fast, bacterium liquid OD
600reach 1.50, in the time that the consumption of peptone is greater than 3.0g/L, bacterium liquid OD
600be less than 1.50.Therefore, peptone is advisable with 3.0g/L.
The impact of the increment of ammonium nitrate on otitis pseudomonas increases along with the increase of the concentration of ammonium nitrate, after the lucifuge of 20h is cultivated, in the time that the consumption of ammonium nitrate is 0.3g/L (every 1000mL basic medium adds 0.3g), its thalline reproduction speed is very fast, bacterium liquid OD
600reach 1.50, in the time that the consumption of ammonium nitrate is less than 0.3g/L, bacterium liquid OD
600be less than 1.50.Therefore, the consumption of ammonium nitrate is advisable with 0.3g/L.
The impact of the increment of yeast extract paste on otitis pseudomonas, increase along with the increase of the concentration of yeast extract paste, after the lucifuge of 20h is cultivated, in the time that the consumption of yeast extract paste is 0.07g/L (every 1000mL basic medium adds 0.07g), its thalline reproduction speed is very fast, bacterium liquid OD
600reach 1.50, in the time that the consumption of yeast extract paste is greater than 0.07g/L, bacterium liquid OD
600be less than 1.50.Therefore, yeast extract paste is advisable with 0.07g/L.
The impact of the increment of vitamins D on otitis pseudomonas, increase along with the increase of the concentration of vitamins D, after the lucifuge of 20h is cultivated, in the time that the consumption of vitamins D is 0.07g/L (every 1000mL basic medium adds 0.07g), its thalline reproduction speed is very fast, bacterium liquid OD
600reach 1.50, in the time that the consumption of vitamins D is greater than 0.07g/L, bacterium liquid OD
600be less than 1.50.Therefore, vitamins D is advisable with 0.07g/L.
The invention has the beneficial effects as follows:
Utilize the method enrichment culture speed of substratum enrichment culture otitis pseudomonas of the present invention fast, owing to having added the nutritive ingredients such as glucose, sodium bicarbonate, peptone, ammonium nitrate, yeast extract paste and vitamins D in basic medium, for microbial growth not only provides carbon source, nitrogenous source, the energy, and add yeast extract paste and the vitamins D as somatomedin, thereby for otitis pseudomonas has been created a better nutritional condition, otitis pseudomonas growth and breeding speed is promoted rapidly, cultivate after 20h its OD
600reach 1.50 left and right.
Otitis pseudomonas extends vegetative period, and maximum growth amount improves.Along with the increase of incubation time, bacterium liquid OD
600increase gradually, while cultivating 20h, its OD
600be 1.50, bacterium liquid increment reaches climax, the OD during than use base culture base otitis pseudomonas
600increase, its increment maximum value has shifted to an earlier date 9～11 hours, and its breeding is more had to promoter action.
Brief description of the drawings
Fig. 1 is the present invention and conventional medium, in substratum enrichment culture process, and optical density(OD) OD
600variation diagram.
In figure, a represents conventional medium, and b represents the substratum of enrichment otitis pseudomonas of the present invention.
Embodiment
Embodiment 1
(1) a kind of preparation of enrichment otitis pseudomonas substratum
Basic medium: dipotassium hydrogen phosphate 3.8g, potassium primary phosphate 1.0g, sodium-chlor 1.0g, ammonium chloride 0.1g, magnesium sulfate 0.2g, distilled water 1000mL.
In above-mentioned basic medium, add 3.0g glucose, 0.3g sodium bicarbonate, 3.0g peptone, 0.3g ammonium nitrate, 0.07g yeast extract paste and 0.07g vitamins D.
(2) cultivation of otitis pseudomonas
The pH of above-mentioned substratum is adjusted to 7.0,121 DEG C of sterilizing 30min.Otitis pseudomonas is inoculated in substratum by 1% inoculum size, and vibration mixes, and is placed in constant incubator and cultivates, 30 DEG C of temperature.
Enrichment culture result: after constant temperature culture 18h, bacterium liquid OD
600be 1.40, after constant temperature culture 20h, reach maximum growth amount, bacterium liquid OD
600be 1.50, make the maximum growth amount of otitis pseudomonas shift to an earlier date 10h.
Embodiment 2
(1) a kind of preparation of enrichment otitis pseudomonas substratum
Basic medium: dipotassium hydrogen phosphate 3.8g, potassium primary phosphate 1.0g, sodium-chlor 1.0g, ammonium chloride 0.1g, magnesium sulfate 0.2g, distilled water 1000mL.
In above-mentioned basic medium, add 1.0g glucose, 0.1g sodium bicarbonate, 1.0g peptone, 0.1g ammonium nitrate, 0.05g yeast extract paste and 0.05g vitamins D.
(2) cultivation of otitis pseudomonas
The pH of above-mentioned substratum is adjusted to 7.0,121 DEG C of sterilizing 30min.Otitis pseudomonas is inoculated in substratum by 1% inoculum size, and vibration mixes, and is placed in constant incubator and cultivates, 30 DEG C of temperature.
Enrichment culture result: after illumination cultivation 18h, bacterium liquid OD
600be 1.30, lucifuge is cultivated after 21h, reaches maximum growth amount, bacterium liquid OD
600be 1.50, make the maximum growth amount of Acinetobacter bauamnnii shift to an earlier date 9h.
Embodiment 3
(1) a kind of preparation of enrichment otitis pseudomonas substratum
Basic medium: dipotassium hydrogen phosphate 3.8g, potassium primary phosphate 1.0g, sodium-chlor 1.0g, ammonium chloride 0.1g, magnesium sulfate 0.2g, distilled water 1000mL.
In above-mentioned basic medium, add 5.0g glucose, 0.5g sodium bicarbonate, 5.0g peptone, 0.5g ammonium nitrate, 0.1g yeast extract paste and 0.1g vitamins D.
(2) cultivation of otitis pseudomonas
The pH of above-mentioned substratum is adjusted to 7.0,121 DEG C of sterilizing 30min.Otitis pseudomonas is inoculated in substratum by 1% inoculum size, and vibration mixes, and is placed in constant incubator and cultivates, 30 DEG C of temperature.
Enrichment culture result: lucifuge is cultivated after 20h, reaches maximum growth amount, bacterium liquid OD
600be 1.50, make the maximum growth amount of otitis pseudomonas shift to an earlier date 9h～11h.
Comparative example
Get respectively the basic medium in substratum and the published embodiment 1 of enrichment otitis pseudomonas in embodiment 1, the pH that regulates respectively substratum is 7,121 DEG C of sterilizing 30min.Otitis pseudomonas is inoculated in respectively in two kinds of substratum by 1% inoculum size, and vibration mixes, and is placed in constant incubator and cultivates, 30 DEG C of temperature.Lucifuge is cultivated 40h, measures optical density(OD) OD in substratum enrichment culture process
600change as shown in Figure 1.From Fig. 1, a is known, uses conventional base culture base otitis pseudomonas, bacterium liquid OD when 30h left and right
600be 0.985; From Fig. 1, b is known, uses culture medium culturing otitis pseudomonas of the present invention, along with the increase of incubation time, and bacterium liquid OD
600increase gradually, cultivate 20h left and right, its OD
600be 1.50, substratum of the present invention make otitis pseudomonas reach maximum growth amount time advance 9h～11h, and to its breeding and fast culture more have promoter action.
Claims (3)
1. the substratum of an enrichment otitis pseudomonas, it is characterized in that, comprise basic medium, glucose, sodium bicarbonate, peptone, ammonium nitrate, yeast extract paste and vitamins D, every 1000mL basic medium adds glucose 1.0～5.0g, sodium bicarbonate 0.1-0.5g, peptone 1.0～5.0g, ammonium nitrate 0.1-0.5g, yeast extract paste 0.05～0.1g and vitamins D 0.05～0.1g.
2. the substratum of enrichment otitis pseudomonas according to claim 1, is characterized in that: every 1000mL basic medium adds glucose 3.0g, sodium bicarbonate 0.3g, peptone 3.0g, ammonium nitrate 0.3g, yeast extract paste 0.07g and vitamins D 0.07g.
3. one kind is utilized the method for the substratum enrichment culture otitis pseudomonas described in claim 1 or 2, it is characterized in that: the pH of the substratum of the enrichment otitis pseudomonas described in claim 1 or 2 is adjusted to 7.0～7.2,121 DEG C of sterilizing 30min, otitis pseudomonas is inoculated in by 1% inoculum size in the substratum of enrichment otitis pseudomonas, vibration mixes, and in temperature is the constant incubator of 30 DEG C, cultivates.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104480045B (en) * | 2014-12-17 | 2017-07-25 | 深圳市深港产学研环保工程技术股份有限公司 | One plant of otitis pseudomonas strains and its application |
| CN107523523A (en) * | 2017-09-30 | 2017-12-29 | 中国科学院成都生物研究所 | One plant of otitis pseudomonad and its application of degradation of feather production oligopeptides |
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| CN103525730A (en) * | 2013-10-14 | 2014-01-22 | 青岛蔚蓝生物集团有限公司 | Pseudomonas otitidis strain and application thereof |
| CN103695355A (en) * | 2013-12-23 | 2014-04-02 | 华南农业大学 | Pseudomonas otitidis H3 strain and application thereof |
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2014
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| CN103497909A (en) * | 2013-08-27 | 2014-01-08 | 中国科学院成都生物研究所 | Bacterial strain JY9 and use thereof |
| CN103525730A (en) * | 2013-10-14 | 2014-01-22 | 青岛蔚蓝生物集团有限公司 | Pseudomonas otitidis strain and application thereof |
| CN103695355A (en) * | 2013-12-23 | 2014-04-02 | 华南农业大学 | Pseudomonas otitidis H3 strain and application thereof |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104480045B (en) * | 2014-12-17 | 2017-07-25 | 深圳市深港产学研环保工程技术股份有限公司 | One plant of otitis pseudomonas strains and its application |
| CN107523523A (en) * | 2017-09-30 | 2017-12-29 | 中国科学院成都生物研究所 | One plant of otitis pseudomonad and its application of degradation of feather production oligopeptides |
| CN107523523B (en) * | 2017-09-30 | 2020-08-21 | 中国科学院成都生物研究所 | Pseudomonas otitis and application thereof in degrading feather to produce oligopeptide |
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