CN106635895A - Lower-serum and efficient culture medium of mycoplasma ovipneumoniae and preparation method of lower-serum and efficient culture medium - Google Patents
Lower-serum and efficient culture medium of mycoplasma ovipneumoniae and preparation method of lower-serum and efficient culture medium Download PDFInfo
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Abstract
The invention discloses a lower-serum and efficient culture medium of mycoplasma ovipneumoniae. The lower-serum and efficient culture medium is prepared from the following components: MEM, sodium pyruvate, glucose, Hank's liquid, fresh yeast leaching liquid, L-glutamine, L-cysteine, lactalbumin hydrolysate, calf thymus DNA (Deoxyribonucleic Acid), insulin, transferrin, penicillin, horse serum, phenol red and de-ionized water; the invention provides a preparation method of the lower-serum and efficient culture medium. The lower-serum and efficient culture medium of the mycoplasma ovipneumoniae, disclosed by the invention, has the beneficial effects that the content of the serum is only 5 percent and is 1/4 of the content of the serum in a culture medium in the prior art; the titer of mycoplasma ovipneumoniae semi-finished-product bacterium liquid prepared by the method reaches 10<10>CCU/ml and is obviously higher than that of the culture medium in the prior art. According to the lower-serum and efficient culture medium of the mycoplasma ovipneumoniae, disclosed by the invention, the sensitive stress response to a goat body, caused by heterologous serum, is alleviated, and the biosafety is improved; the titer of bacterium liquid of living bacteria is also improved and the production cost is reduced.
Description
Technical field
The present invention relates to veterinary formulations technical field, and in particular to a kind of low serum efficient culture medium of mycoplasma ovine pneumoniae
And preparation method thereof.
Background technology
Mycoplasma ovine pneumoniae（Mycoplasma ovipneumoniae, Mo）The chromic fibrous of sheep and goat can be caused
Pneumonia, Clinical symptoms be mainly shown as cough, pant, have a running nose, becoming thin, anaemia, growth retardation etc..Mackay in 1963
Deng first from Scotland sheep disease lung in isolate mycoplasma ovine pneumoniae.Mycoplasma ovine pneumoniae main infection sheep and mountain
Sheep, the morbidity report of the sheep and goat for having mycoplasma ovine pneumoniae to cause in many countries in the whole world at present, supports to the world
The huge economic losses that sheep industry is caused.Chinese scholar in 1978 introduces border area Leicester and plants sheep in Sichuan Province from New Zealand first
Mycoplasma ovine pneumoniae is isolated in offspring lamb.Subsequently there are goat and sheep infection pneumonia of sheep in multiple provinces and cities of China in succession
The report of substance morbidity, the disease is widely current in China, becomes a kind of important pathogen of harm China sheep husbandry.
Vaccine immunity is the important means that the infection of prevention and control mycoplasma ovine pneumoniae occurs.The sheep lung of prevention and control at present
Scorching mycoplasma infection relies primarily on inactivated vaccine.Culture medium is the key of production of vaccine technique.Current mycoplasma ovine pneumoniae
Culture medium mainly has improvement KM2 culture mediums, improvement Thiaucourt's culture mediums, TSB-1 culture mediums etc..Prior art culture medium
Culture mycoplasma ovine pneumoniae has that incubation time length, viable bacteria titre is low, fertility.Additionally, prior art training
Serum content is commonly 20% in foster base, and vaccine prepared by high-load serum undoubtedly increased allogeneic serum should to the allergy of sheep body
Swash reaction, the immune effect of final impact vaccine, while also increasing seedling cost.If reducing prior art culture medium merely
Middle serum content, can cause to cultivate low 10～100 times or so of sheep mycoplasma antigen titre, both not reach antigen-agent needed for immunity
Amount, need to improve cycles of concentration again, actually increased production cost.Therefore, the low serum high-efficient culture of mycoplasma ovine pneumoniae is developed
Base becomes the major issue for being badly in need of solving in mycoplasma ovine pneumoniae vaccine research and production.
The content of the invention
The purpose of the present invention is aiming at above-mentioned defect of the prior art, there is provided a kind of low blood of mycoplasma ovine pneumoniae
Clear efficient culture medium and preparation method thereof.
To achieve these goals, the technical scheme of present invention offer is：A kind of low serum of mycoplasma ovine pneumoniae is efficient
Culture medium, is made up of per the low serum efficient culture mediums of 1000ml basal medium and auxiliary culture medium；
Contain following components in the basal medium：
（1）The g of MEM 6.0～7.0,
（2）The g of Sodium Pyruvate 5.0～6.0,
（3）The g of glucose 5.0～6.0,
（4）Mass concentration is 1% phenol red 2.0～2.5 ml,
（5）The ml of Hank ' s liquid 500,
（6）The ml of deionized water 300；
Contain following components in the auxiliary culture medium：
（7）Mass concentration is 25% ml of fresh yeast leachate 30～50,
（8）Mass concentration is 3% ml of Glu 16～17,
（9）Mass concentration is 10% ml of Cys 4.0～6.0,
（10）Mass concentration is 15% ml of lactoalbumin hydrolysate 32.0～35.0,
（11）0.8～the 1.5ml of transferrins of 10 mg/ml,
（12）The ml of insulin 0.8～1.5 of 10 mg/ml,
（13）The ml of calf thymus DNA 8.0～10.0 of 2mg/ml,
（14）The ml of penicillin 0.25 of 200000 IU/ml,
（15）The ml of sterile horse blood serum 50.
Second object of the present invention there is provided a kind of system of the low serum efficient culture medium of above-mentioned mycoplasma ovine pneumoniae
Preparation Method, comprises the following steps：
1）The preparation of basal medium：
In taking above-mentioned basal medium component（1）-（5）300ml deionized waters are added one by one（6）In, it is sufficiently mixed, 115
It is standby room temperature to be cooled to after DEG C autoclaving 20min；
2）The preparation of auxiliary culture medium：
Learn from else's experience the above-mentioned of 0.22 micron membrane filter EK（7）-（15）Composition, is sufficiently mixed, and obtains final product auxiliary culture medium；
3）Mixing constant volume：
Under aseptic condition, by step 1）The basal medium for obtaining and step 2）The auxiliary culture medium mixing for obtaining, uses aqua sterilisa
Polishing is settled to 1000ml, with the 1M NaOH of sterilizing（4 g are dissolved in 100 ml deionized waters）Adjust pH value to 7.2-7.4, fully shake
Even, rearmounted 4 DEG C of packing is saved backup.
Third object of the present invention there is provided a kind of low serum efficient culture medium of above-mentioned mycoplasma ovine pneumoniae in training
Application in foster mycoplasma ovine pneumoniae.
Fourth object of the present invention there is provided a kind of low serum efficient culture medium of above-mentioned mycoplasma ovine pneumoniae in system
Application in standby mycoplasma ovine pneumoniae vaccine antigen.
Beneficial effects of the present invention are：The low serum efficient culture medium of mycoplasma ovine pneumoniae of the present invention is according to sheep lung
The growth characteristics of scorching mycoplasma, are entered by the combination to many nutrition compositions such as different carbon sources, nitrogen source, protein, inorganic salts
Screening is gone, pH value, osmotic pressure, ionic strength to culture medium etc. are compared analysis, investigated and be adapted to culture sheep lung
The low serum efficient culture medium of scorching mycoplasma.MEM and Hank ' s liquid are mycoplasma growth basal liquids in the culture medium；In culture medium
Glucose for mycoplasma ovine pneumoniae growth carbon source and energy are provided；Sodium Pyruvate can be used as the replacement carbon of mycoplasma growth
Source；Nitrogen source, electrolytes and minerals by adding fresh yeast leachate, for needed for mycoplasma ovine pneumoniae grows offer
Etc. nutrient content；Addition Glu can promote microbial metabolism, promote protein synthesis；Addition Cys and hydrolysis
Lactoprotein provides suitable amino acid and growth factor for mycoplasma ovine pneumoniae growth；Insulin not only have promote glycogen with
The synthesis of aliphatic acid, and the synthesis of protein, lipid and RNA can be promoted；Transferrins is that microorganism obtains trace element
Important way, plays a role with insulin is promoted；By adding horse serum, provide needed for growth to mycoplasma ovine pneumoniae
Cholesterol and required saturation or unrighted acid；Varied bacteria growing can be suppressed by adding penicillin, to mycoplasma unrestraint
Effect, can avoid culture medium pollution and Extending culture base holding time；Addition is phenol red as pH, can determine whether mycoplasma
Upgrowth situation.In the formula of the culture medium in addition to fundamental component, fresh yeast leachate, L- paddy are also added in the medium
The compositions such as glutamine, Cys, lactoalbumin hydrolysate, calf thymus DNA, transferrins, insulin, the addition of mentioned component
The viable bacteria titre of mycoplasma ovine pneumoniae can be significantly improved；Jing is repeatedly it is experimentally confirmed that viable bacteria titre is 10 before being not added with7～108
CCU/ml, viable bacteria titre is up to 10 after addition10CCU/ml.And the best advantage is that low serum contains in culture medium
Amount and culture mycoplasma ovine pneumoniae viable bacteria titre are high.Culture medium horse serum consumption of the present invention is only 5%, is prior art culture
The 1/4 of base serum content；The mycoplasma ovine pneumoniae semi-finished product bacterium solution titre prepared with the inventive method is up to 1010CCU/ml, changes
Good KM2 medium cultures mycoplasma ovine pneumoniae viable bacteria titre is 107CCU/ml, improvement Thiaucourt's culture mediums and TSB-
1 medium culture mycoplasma ovine pneumoniae viable bacteria titre is 108～109CCU/ml, using low serum efficient culture medium of the invention
Culture mycoplasma ovine pneumoniae viable bacteria titre apparently higher than improvement KM2 culture mediums, improvement Thiaucourt's culture mediums and
TSB-1 culture mediums.Low serum efficient culture medium significantly reduces allergy stress reaction of the alloplasm serum to sheep body in vaccine, carries
High bio-safety；Mycoplasma ovine pneumoniae viable bacteria titre is cultivated simultaneously high, reduce production of vaccine cost, be pneumonia of sheep
The development of substance high-quality vaccine is laid a good foundation.
Description of the drawings
Fig. 1 is shown as growing state of the mycoplasma ovine pneumoniae in low serum efficient culture medium of the invention.
Specific embodiment
Preparation source used of the invention：
MEM：GIBCO Products, article No. 61100-087.
Glucose：Beijing Chemical Plant's product.
Sodium Pyruvate：AMRESCO Products, article No. is 0342.
Glu：Sigma Products, article No. is V900419.
Cys：AMRESCO Products, article No. is J994.
Lactoalbumin hydrolysate：Beijing Suo Laibao Science and Technology Ltd product, article No. is L8100.
Transferrins：Sigma Products, article No. is T8158.
Insulin：HUMOBIO Products, article No. is HAK4570.
Calf thymus DNA：Sigma Products, article No. is D1501.
Penicillin：For Shanghai Gongyi Veterinary Medicines Plant's product.
Horse serum：Hyclone Products, article No. is SH30074.03.
It is phenol red：Sigma Products, article No. is P3532.
Mass concentration is 1% phenol red preparation：Phenol red 1.0 g is weighed, in putting glass mortar, 0.1M NaOH is added dropwise over
（0.4 g is dissolved in 10 ml deionized waters）, it is ground to and is completely dissolved.By in the ml measuring bottles of phenol red suction 100 of dissolving, deionization is used
Water is carefully washed and phenol red liquid is remained in lower mortar deionized water is added into measuring bottle, finally to 100 ml.
The preparation of Hank ' s liquid：Weigh 5.0 g sodium chloride（NaCl）, 0.4 g potassium chloride（KCl）, 1.6 g phosphoric acid hydrogen two
Sodium（Na2HPO4。12H2O）, 0.1 g potassium dihydrogen phosphates（KH2PO4), 0.2 g magnesium sulfate（MgSO4.7H2O）, 10 g glucose, use
800 ml deionized water dissolvings are simultaneously settled to 1000 ml, 115 DEG C of autoclavings 20 minutes, put 4 DEG C and save backup.
Mass concentration is the preparation of 25% fresh yeast leachate：The g of yeast cake 500 is taken, the ml of deionized water 2000 is added
In, stirring and dissolving uses concentrated hydrochloric acid：Deionized water is with=1:1（Volume ratio）PH value is adjusted to 4.5-5.0,80 DEG C of water-baths（Temperature in bottle）
30 minutes, 3000 revs/min were centrifuged 20 minutes, take supernatant.By supernatant with 1 M NaOH（4 g are dissolved in 100 ml deionizations
Water）Adjust pH value to 7.8-8.0, boil, put and filtered with double-layer filter paper after room temperature is cooled, then mend deionized water to 2000 ml, -20
DEG C save backup.
Embodiment 1：
1st, the preparation of culture medium of the present invention：
Basal medium：
（1）MEM 6.5 g
（2）The g of Sodium Pyruvate 6.0
（3）The g of glucose 6.0
（4）Mass concentration is 1% phenol red 2.5 ml
（5）The ml of Hank ' s liquid 500
（6）The ml of deionized water 300.
Auxiliary culture medium：
（7）Mass concentration is 25% ml of fresh yeast leachate 30
（8）Mass concentration is 3% ml of Glu 16
（9）Mass concentration is 10% ml of Cys 5.0
（10）Mass concentration is the ml of 15% lactoalbumin hydrolysate 32.5
（11）Transferrins（10 mg/ml）1.0 ml
（12）Insulin（10 mg/ml）1.0 ml
（13）Calf thymus DNA（2mg/ml） 10.0 ml
（14）Penicillin (200,000 IU/ml) 0.25 ml
（15）The ml of sterile horse blood serum 50
By in basal medium（1）-（5）Composition dissolves in one by one 300ml deionized waters（6）In, 115 DEG C of autoclaving 20min.Treat
After being cooled to room temperature, the auxiliary culture medium of the micron membrane filter EKs of Jing 0.22 is aseptically added（7）-（15）Composition,
Mixing, with aqua sterilisa polishing 1000ml is settled to, and adjusts pH value to 7.4 with the 1M NaOH of sterilizing, is dispensed after fully shaking up, and puts 4 DEG C
Save backup.
Embodiment 2：
The preparation of culture medium of the present invention：
Basal medium：
（1）MEM 6.0 g
（2）The g of Sodium Pyruvate 6.0
（3）The g of glucose 6.0
（4）Mass concentration is 1% phenol red 2.5 ml
（5）The ml of Hank ' s liquid 500
（6）The ml of deionized water 300.
Auxiliary culture medium：
（7）Mass concentration is 25% ml of fresh yeast leachate 35
（8）Mass concentration is 3% ml of Glu 17
（9）Mass concentration is 10% ml of Cys 5.0
（10）Mass concentration is 15% ml of lactoalbumin hydrolysate 35.0
（11）Transferrins（10 mg/ml）1.0 ml
（12）Insulin（10 mg/ml）0.8 ml
（13）Calf thymus DNA（2mg/ml） 10.0 ml
（14）Penicillin (200,000 IU/ml) 0.25 ml
（15）The ml of sterile horse blood serum 50
By in basal medium（1）-（5）Composition dissolves in one by one 300ml deionized waters（6）In, 115 DEG C of autoclaving 20min.Treat
After being cooled to room temperature, the auxiliary culture medium of the micron membrane filter EKs of Jing 0.22 is aseptically added（7）-（15）Composition,
Mixing, with aqua sterilisa polishing 1000ml is settled to, and adjusts pH value to 7.4 with the 1M NaOH of sterilizing, is dispensed after fully shaking up, and puts 4 DEG C
Save backup.
Embodiment 3：
Using culture medium of the present invention, improvement KM2 culture mediums, improvement Thiaucourt's culture mediums, TSB-1 culture mediums to sheep lung
Scorching mycoplasma Y98 strains are compared test：
1st, the preparation of culture medium of the present invention
Basal liquid：
（1）MEM 6.0 g
（2）The g of Sodium Pyruvate 5.0
（3）The g of glucose 6.0
（4）Mass concentration is 1% phenol red 2.5 ml
（5）The ml of Hank ' s liquid 500
（6）The ml of deionized water 300.
Auxiliary culture medium：
（7）Mass concentration is 25% ml of fresh yeast leachate 30
（8）Mass concentration is 3% ml of Glu 16.7
（9）Mass concentration is 10% ml of Cys 5.0
（10）Mass concentration is 15% ml of lactoalbumin hydrolysate 33.4
（11）Transferrins（10 mg/ml）1.0 ml
（12）Insulin（10 mg/ml）1.0 ml
（13）Calf thymus DNA（2mg/ml） 10.0 ml
（14）Penicillin (200,000 IU/ml) 0.25 ml
（15）The ml of sterile horse blood serum 50
By in basal medium（1）-（5）Composition dissolves in one by one 300ml deionized waters（6）In, 115 DEG C of autoclaving 20min.Treat
After being cooled to room temperature, the auxiliary culture medium of the micron membrane filter EKs of Jing 0.22 is aseptically added（7）-（15）Composition,
Mixing, with aqua sterilisa polishing 1000ml is settled to, and adjusts pH value to 7.4 with the 1M NaOH of sterilizing, is dispensed after fully shaking up, and puts 4 DEG C
Save backup.
2nd, the preparation of Thiaucourt's culture mediums is improved（1000ml）
Basal liquid：
The g of PPLO meat soups 21.0
The ml of deionized water 700.
115 DEG C of min of autoclaving 20；
Culture medium：
Basal liquid 700ml
The ml of 50% Sodium Pyruvate 4.0
The ml of 50% glucose 2.0
Mass concentration is 1% phenol red 2.5 ml
Mass concentration is 25% ml of fresh yeast leachate 100
Penicillin（200000 IU/ml）1.0 ml
The ml of 10% thaliium acetate 1.0
The ml of sterile horse blood serum 200
It is standby that the 1M NaOH of mixing sterilizing adjust pH value to dispense to the micron membrane filter EKs of 7.4, Jing 0.22.
3rd, TSB-1 culture mediums（1000ml）
Pancreas peptone soybean broth 30g
The g of lactose 10
DNA 0.02g
Mass concentration is 1% phenol red 2.5 ml
Penicillin（200000 IU/ml）1.0 ml
The ml of 10% thaliium acetate 1.0
The ml of aseptic Swine serum 200
Deionized water is settled to 1000ml, and pH value is adjusted to the micron membrane filter EKs of 7.4, Jing 0.22 with the 1M NaOH of sterilizing
Packing is standby.
4th, KM2 culture mediums are improved（500ml）
The ml of 1.7% lactoalbumin hydrolysate Hank ' s liquid 150
MEM 2.5 g
The g of glucose 2.0
The g of Sodium Pyruvate 1.0
Mass concentration is 1% phenol red 1.25 ml
Mass concentration is 25% ml of fresh yeast leachate 10
Penicillin（200000 IU/ml）0.5 ml
The ml of 10% thaliium acetate 0.5
The ml of sterile horse blood serum 100
Deionized water is settled to 500ml, and pH value is adjusted to the micron membrane filter EKs of 7.4, Jing 0.22 with the 1M NaOH of sterilizing
Packing is standby.
5th, the culture of mycoplasma ovine pneumoniae is by mycoplasma ovine pneumoniae Y98 strains（Protect purchased from Chinese veterinary microorganism bacterial classification
Tibetan center CVCC, numbering CVCC384）Low serum efficient culture medium of the invention is inoculated with respectively（Serum content is 5%）, improvement KM2 training
Foster base（Serum content is 20%）, improvement Thiaucourt's culture mediums（Serum content is 20%）, TSB-1 culture mediums（Serum contains
Measure as 20%）, after seed subculture rejuvenation, respectively in the ratio inoculation correspondence culture medium of 10% (V/V), 37 DEG C are incubated, when
Its colour changed into yellow, the pH value of culture medium when 7.4 are down to 6.5～6.8, aseptic taking-up culture.
6th, viable bacteria titre（CCU）Determine.Method is as follows：12 test tubes are taken, often pipe plus the correspondence ml of culture medium 4.5, the 1st
The well-grown mycoplasma ovine pneumoniae cultures of 0.5 ml are added in pipe, is fully mixed with oscillator, the pipette for renewing, from
1st pipe is drawn 0.5 ml and is added in the 2nd pipe, 10 times is carried out successively and is diluted to the 11st pipe, and 0.5 ml cultures are discarded in the 11st pipe
Liquid；The dilution factor for obtaining nutrient solution is respectively 10-1-10-11, the 12nd pipe is correspondence culture medium control.Developmental tube sets 3 repetitions.Will
Test tube puts quiescent culture in 37 DEG C of constant incubators, and color change is observed daily, and Continuous Observation 10 days occurs color change most
High dilution is the viable bacteria titre of the culture, uses color changing units（CCU）Represent.Test is repeated 3 times.
7th, result：
Mycoplasma ovine pneumoniae is inoculated with the 4 kinds of secondary growth of culture medium 3 test CCU measurement results and is shown in Table 1.
Table 1
3 times result of the test shows, under similarity condition, using improvement KM2 culture mediums, improvement Thiaucourt's culture mediums,
The growth time of TSB-1 medium culture mycoplasma ovine pneumoniaes is 48h, and the growth time of culture medium of the present invention is 24h, than
Prior art incubation time shortens 1/2.It is equal using improvement KM2 medium cultures 3 CCU measurement results of mycoplasma ovine pneumoniae
For 107CCU/ml, 3 CCU measurement results of improvement Thiaucourt's medium cultures mycoplasma ovine pneumoniae are 108～109
3 CCU measurement results of CCU/ml, TSB-1 medium culture mycoplasma ovine pneumoniae are 108 ～109CCU/ml；The present invention is low
Serum efficient culture medium culture mycoplasma ovine pneumoniae, growing state is shown in Fig. 1.Medium culture mycoplasma ovine pneumoniae of the present invention
3 times CCU measurement results are 1010 CCU/ml.As a result show, under similarity condition, using low serum efficient culture medium of the invention
Viable bacteria titre significantly larger than improves KM2 culture mediums, is significantly higher than improvement Thiaucourt's culture mediums and TSB-1 culture mediums.With
Upper result explanation, the mycoplasma ovine pneumoniae culture medium of the present invention has that serum content is low, growth is rapid and viable bacteria titre is high
Feature.
Finally it should be noted that：The preferred embodiments of the present invention are the foregoing is only, the present invention is not limited to,
Although being described in detail to the present invention with reference to the foregoing embodiments, for a person skilled in the art, it still may be used
To modify to the technical scheme described in foregoing embodiments, or equivalent is carried out to which part technical characteristic.
All any modification, equivalent substitution and improvements within the spirit and principles in the present invention, made etc., should be included in the present invention's
Within protection domain.
Claims (4)
1. the low serum efficient culture medium of a kind of mycoplasma ovine pneumoniae, it is characterised in that per the low serum efficient culture mediums of 1000ml
It is made up of basal medium and auxiliary culture medium；
Contain following components in the basal medium：
（1）The g of MEM 6.0～7.0,
（2）The g of Sodium Pyruvate 5.0～6.0,
（3）The g of glucose 5.0～6.0,
（4）Mass concentration is 1% phenol red 2.0～2.5 ml,
（5）The ml of Hank ' s liquid 500,
（6）The ml of deionized water 300；
Contain following components in the auxiliary culture medium：
（7）Mass concentration is 25% ml of fresh yeast leachate 30～50,
（8）Mass concentration is 3% ml of Glu 16～17,
（9）Mass concentration is 10% ml of Cys 4.0～6.0,
（10）Mass concentration is 15% ml of lactoalbumin hydrolysate 32.0～35.0,
（11）0.8～the 1.5ml of transferrins of 10 mg/ml,
（12）The ml of insulin 0.8～1.5 of 10 mg/ml,
（13）The ml of calf thymus DNA 8.0～10.0 of 2mg/ml,
（14）The ml of penicillin 0.25 of 200000 IU/ml,
（15）The ml of sterile horse blood serum 50.
2. the preparation method of the low serum efficient culture medium of a kind of mycoplasma ovine pneumoniae according to claim 1, its feature
It is to comprise the following steps：
1）The preparation of basal medium：
In taking the basal medium component described in claim 1（1）-（5）300ml deionized waters are added one by one（6）In, fully
Mixing, is cooled to room temperature standby after 115 DEG C of autoclaving 20min；
2）The preparation of auxiliary culture medium：
Learn from else's experience described in the claim 1 of 0.22 micron membrane filter EK（7）-（15）Composition, is sufficiently mixed, and obtains final product auxiliary
Culture medium；
3）Mixing constant volume：
Under aseptic condition, by step 1）The basal medium for obtaining and step 2）The auxiliary culture medium mixing for obtaining, uses aqua sterilisa
Polishing is settled to 1000ml, adjusts pH value to 7.2-7.4 with the 1M NaOH of sterilizing, fully shakes up, and rearmounted 4 DEG C of packing is saved backup.
3. a kind of low serum efficient culture medium of mycoplasma ovine pneumoniae according to claim 1 is in culture pneumonia of sheep original
Application in body.
4. a kind of low serum efficient culture medium of mycoplasma ovine pneumoniae according to claim 1 is preparing pneumonia of sheep original
Application in body vaccine antigen.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110938560A (en) * | 2019-11-19 | 2020-03-31 | 成都天邦生物制品有限公司 | Rapid and efficient mycoplasma hyopneumoniae culture medium and preparation method thereof |
| CN111635876A (en) * | 2020-06-16 | 2020-09-08 | 武汉科前生物股份有限公司 | Mycoplasma hyopneumoniae culture medium and preparation method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103074246A (en) * | 2012-08-31 | 2013-05-01 | 南京天邦生物科技有限公司 | Low serum efficiency mycoplasma gallisepticam attenuated strain culture medium and preparation method thereof |
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| CN103074246A (en) * | 2012-08-31 | 2013-05-01 | 南京天邦生物科技有限公司 | Low serum efficiency mycoplasma gallisepticam attenuated strain culture medium and preparation method thereof |
| CN103110583A (en) * | 2013-03-05 | 2013-05-22 | 西南民族大学 | Preparation technology of mycoplasma ovipneumoniae inactivated vaccine |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110938560A (en) * | 2019-11-19 | 2020-03-31 | 成都天邦生物制品有限公司 | Rapid and efficient mycoplasma hyopneumoniae culture medium and preparation method thereof |
| CN110938560B (en) * | 2019-11-19 | 2023-08-18 | 成都史纪生物制药有限公司 | Rapid and efficient mycoplasma hyopneumoniae culture medium and preparation method thereof |
| CN111635876A (en) * | 2020-06-16 | 2020-09-08 | 武汉科前生物股份有限公司 | Mycoplasma hyopneumoniae culture medium and preparation method and application thereof |
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