CN101307293A - Culture collection process - Google Patents
Culture collection process Download PDFInfo
- Publication number
- CN101307293A CN101307293A CNA2008101243255A CN200810124325A CN101307293A CN 101307293 A CN101307293 A CN 101307293A CN A2008101243255 A CNA2008101243255 A CN A2008101243255A CN 200810124325 A CN200810124325 A CN 200810124325A CN 101307293 A CN101307293 A CN 101307293A
- Authority
- CN
- China
- Prior art keywords
- spawn
- bacterial classification
- culture medium
- semisolid
- aseptic
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention provides a spawn preservation method adopting semisolid low-temperature air-tight culture. A spawn is inoculated in an aseptic semisolid culture medium which is vertically stored at an ambient temperature of between 6 and 8 DEG C after aseptic liquid paraffin is added into the culture medium. The spawn is staphylococcus aureus, escherichia coli, pseudomonas aeruginosa, bacillus subtilis, clostridium orogenes, aspergillus niger or candida albicans. The specific process is as follows: preparing the semisolid culture medium, carrying out sub-package and sterilization, and then culturing for 2 to 3 days, inoculating the spawn into the semisolid culture medium in a mode of stab inoculation after asepsis is ensured, with the number of stab inoculation points parallel with the plane of the culture medium being no less than 3; pouring the aseptic liquid paraffin with a height of 1 to 2 cm into the culture medium which is sealed by a sealing film and is stored in a refrigerator at a temperature of between 6 and 8 DEG C. The method can not only prolong the spawn preservation time but also keep a relatively stable spawn concentration, thereby saving much cost for production and test, reducing the treatment and discharge of rejected material after spawn use, and reducing environmental hazard.
Description
Technical field
The present invention relates to a kind of culture collection process.
Background technology
Stipulate according to the Pharmacopoeia of the People's Republic of China, the requisite of the positive contrast of use of bacterial classification in the microbial limit check of medicine, the steriling test, it is a kind of special reference substance, and it is essential to be similarly in microbial limit check, the checking of steriling test methodology institute.Therefore for a long time, the use of bacterial classification and preservation always are the emphasis of Microbiological Lab.Normally used bacterial classification has 7 kinds, is respectively: streptococcus aureus, escherichia coli, Pseudomonas aeruginosa, subtilis, clostridium sporogenes, aspergillus niger, white chain pearl bacterium.At present, domestic drug manufacturing enterprise uses cell preservation technique many years ago always, and is because this method utilization for many years, more stable reliable.This method is: the bacterial classification of buying the low temperature slant preservation from medicine inspecting institute normally uses.The bacterial classification of buying is generally third generation bacterial classification, and the use of bacterial classification can not surpass for 5 generations.Because the shelf time of this method for preserving bacterial classification is short, the bacterial classification instability of preservation.Therefore will use certain density bacterial classification need go down to posterity once every month, use promptly that the third generation just became for the 4th generation after one month, the bacterial classification that is to say purchase only can use will be bought in 3 months again.Every low temperature slant preservation bacterial classification is bought about about 100 circles of the price of coming from province medicine inspecting institute, and 7 bacterial classifications are circle surplus in the of 700, nearly 3000 circles of the drug manufacturing enterprise's cost of every year aspect bacterial classification.The waste very of such method, same bacterial classification use the processing of back waste also to want exactissima diligentia, will pollute environment because of carelessness, even can cause not having the people of relevant knowledge sick, very serious of consequence.Relevant both at home and abroad simultaneously documentation is less, can't solve present problem.
Summary of the invention
The invention provides a kind of culture collection process, make the concentration of bacterial classification keep relative stable when can prolong shelf time of bacterial classification, be produce, test saves cost, reduces processing and discharging that bacterial classification uses the back waste.
The invention provides a kind of culture collection process, adopt semi-solid, low temperature seal cultured method: bacterial classification inoculation in aseptic semisolid medium, is poured in the envrionment temperature of vertically depositing 6～8 ℃ behind the sterile liquid paraffin and preserved.
Described bacterial classification is streptococcus aureus, escherichia coli, Pseudomonas aeruginosa, subtilis, clostridium sporogenes, aspergillus niger or white chain pearl bacterium.
Detailed process is as follows:
(1) according to Pharmacopoeia of the People's Republic of China regulation, add purified water and make semisolid medium in the dehydrated medium of following prescription by mass, packing, sterilization were cultivated 2-3 days then, determined aseptic:
Peptone 10
Sodium-chlor 5
Beef extract powder 3
Agar 4
(2) with the bacterial classification percutaneous puncture-inoculation in semisolid medium, be parallel to substratum plane percutaneous puncture-inoculation and be no less than 3 points.
(3) pour the high sterile liquid paraffin of 1～2cm into,, vertically deposit in 6～8 ℃ the envrionment temperature and preserve with sealing film phonograph seal.
Discover by arrangement, most of bacterial strains can't be all dead immediately under the situation of anoxic, low temperature, low nutriment, but there is the bacterial classification more than 80% to be in dormant state, in case living environment is changed into the condition that is fit to its existence, will recover immediately, through about 18 hours time, the concentration of its bacteria suspension can return to about 95% of the former bacterial concentration of dormancy.Utilize this kind physilogical characteristics of bacterial classification can solve bacterial classification shelf time weak point, the bacterial classification problem of unstable of preservation.Find through test of many times, adopt the present invention the shelf time of bacterial classification can be prolonged, the concentration of bacterial classification keeps relative stablizing simultaneously, and the concentration of preserving 12 months bacteria suspension still can reach about 90% of stoste, and the bacteria suspension concentration of preserving 18 months is also at more than 85% of stoste.This kind method is simple, injury to bacterial classification is very low, the probability of the successful recovery of bacterial classification is very big, and the time less of cost when bacterial classification uses, need about 20 hours from recovery to only using, and low temperature, slant strains preservation rule need about 40 hours, have improved whole production efficient, for produce, test saved a large amount of costs.We are example with the drug manufacturing enterprise: if each bacterial classification was preserved 12 months with this method, on average needing bacterial classification is 7 kinds.Method originally is annual need buy bacterial classification 4 times, spends nearly 3000 circles.After adopting this method, the annual bacterial classification of purchase that only needs only needs about 700 circles, can save nearly 2100 circles aspect the bacterial classification purchase.Save production cost nearly 70%.The present invention simultaneously reduced processing and the discharging that bacterial classification uses the back waste, reduced the discharging of waste water, refuse, reduced the harm to environment.
Embodiment
In dehydrated medium, add purified water and make semisolid medium.By mass, dehydrated medium is composed as follows:
Peptone 10
Sodium-chlor 5
Beef extract powder 3
Agar 4
Packing, sterilization.Pack in the every test tube substratum of 10～20ml is vertically put into incubator and was cultivated 2～3 days, observes, test, determines to carry out next step operation after aseptic.This step is not destroyed by assorted bacterium for the purity of the bacterial classification that guarantees to inoculate.Ready bacterial classification is a small amount of with the inoculating needle picking, and percutaneous puncture-inoculation is parallel to percutaneous puncture-inoculation 3 points on the circular flat in semisolid medium, determine to be seeded in the substratum to guarantee bacterial classification.Pour the high sterile liquid paraffin of 1～2cm into,, bottleneck is sealed, vertically deposit in 6～8 ℃ the refrigerator and preserve, get final product with sealing film with test tube plug jail.
Recovery 18-24 hour relative bacterial concentration behind each culture presevation different time (i.e. the ratio of Fu Su bacteria suspension concentration and stoste bacteria suspension concentration) is as shown in table 1.
Relative bacterial concentration behind each culture presevation different time of table 1
Claims (6)
1, a kind of culture collection process is characterized in that adopting semi-solid, low temperature seal cultured method: bacterial classification inoculation in aseptic semisolid medium, is poured in the envrionment temperature of vertically depositing 6～8 ℃ behind the sterile liquid paraffin and preserved.
2, culture presevation way as claimed in claim 1 is characterized in that described bacterial classification is streptococcus aureus, escherichia coli, Pseudomonas aeruginosa, subtilis, clostridium sporogenes, aspergillus niger or white chain pearl bacterium.
3, culture presevation way as claimed in claim 1 is characterized in that the bacterial classification percutaneous puncture-inoculation in aseptic semisolid medium, is parallel to substratum plane percutaneous puncture-inoculation and is no less than 3 points.
4, as claim 1 or 2 or 3 described culture presevation ways, it is characterized in that described semisolid medium preparation method is as follows: in the dehydrated medium of following prescription by mass, add purified water and make semisolid medium, packing, sterilization were cultivated 2-3 days then, determined aseptic:
Peptone 10
Sodium-chlor 5
Beef extract powder 3
Agar 4
5, as claim 1 or 2 or 3 described culture presevation ways, the height that it is characterized in that sterile liquid paraffin is 1-2cm.
6,, it is characterized in that pouring into behind the sterile liquid paraffin with sealing film phonograph seal as claim 1 or 2 or 3 described culture presevation ways.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008101243255A CN101307293A (en) | 2008-06-26 | 2008-06-26 | Culture collection process |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008101243255A CN101307293A (en) | 2008-06-26 | 2008-06-26 | Culture collection process |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101307293A true CN101307293A (en) | 2008-11-19 |
Family
ID=40123981
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2008101243255A Pending CN101307293A (en) | 2008-06-26 | 2008-06-26 | Culture collection process |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101307293A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102517240A (en) * | 2012-01-12 | 2012-06-27 | 山东省千佛山医院 | Solid culture medium suitable for co-growing Candida albicans and staphylococcus |
| CN111808753A (en) * | 2020-07-22 | 2020-10-23 | 河北化工医药职业技术学院 | Penicillin bottle preservation method for strains |
-
2008
- 2008-06-26 CN CNA2008101243255A patent/CN101307293A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102517240A (en) * | 2012-01-12 | 2012-06-27 | 山东省千佛山医院 | Solid culture medium suitable for co-growing Candida albicans and staphylococcus |
| CN102517240B (en) * | 2012-01-12 | 2015-08-12 | 山东省千佛山医院 | Be applicable to the solid medium of Candida albicans and staphylococcus syntrophism |
| CN111808753A (en) * | 2020-07-22 | 2020-10-23 | 河北化工医药职业技术学院 | Penicillin bottle preservation method for strains |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Gómez-Rivas et al. | Selective antimicrobial action of chitosan against spoilage yeasts in mixed culture fermentations | |
| CN101974607B (en) | Reagent and kit for detecting sensitivity of culture medium | |
| Mensah et al. | Use of pineapple waste for single cell protein (SCP) production and the effect of substrate concentration on the yield | |
| CN102660461A (en) | Microbial preparation for shortening tobacco fermentation period and application of microbial preparation | |
| Lorenzetti et al. | Alginate/PVA Beads for Levan Production by Z ymomonas mobilis | |
| WO2023142723A1 (en) | Method for fast sterility detection in sterile drug substances | |
| Mohapatra et al. | Production of tannase by the immobilized cells of Bacillus licheniformis KBR6 in Ca‐alginate beads | |
| CN105062891A (en) | Enzyme-containing composition and method for enhancing stability of liquid microbial inoculant | |
| CN101307293A (en) | Culture collection process | |
| Sar et al. | Repeated batch fermentation of immobilized E. coli expressing Vitreoscilla hemoglobin for long-term use | |
| CN104862259A (en) | Complex microbial agent for high-organic-load and medium-temperature biogas fermentation as well as preparation method and application thereof | |
| CN104152373A (en) | Bacterial strain capable of efficiently degrading pendimethalin and application thereof | |
| CN1358838A (en) | Preservation method for live bacterial preparation | |
| CN105494320A (en) | Method for preserving straws and preparing marsh gas | |
| CN102061279A (en) | Method for producing rhodopseudomonas palustris fermentation liquor by high-density fermentation | |
| CN102212492A (en) | Bacillus culture medium and culture method | |
| CN1884509A (en) | Preparation method of hybrid hydrolase formulation for promoting methane yield from fermentation | |
| CN104313116A (en) | Columbia blood agar base culture medium | |
| Chang et al. | Growth and production of cholesterol oxidase by alginate‐immobilized cells of Rhodococcus equi No. 23 | |
| CN105255736A (en) | Preservation method of hypsizigus marmoreus strain | |
| CN201301321Y (en) | Microbial cultivation oxygen increase shaking bottle | |
| CN104651254A (en) | Enterococcus faecalis agar medium and use thereof | |
| CN105347855A (en) | Hypsizygus marmoreus strain medium and preparation method thereof | |
| CN104651448A (en) | Liquid thioglycollate medium and use thereof | |
| CN103525737B (en) | Cultivate enrichment medium and the application thereof of rhodopseudomonas |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Open date: 20081119 |