CN105695374A - Bacillus diluent, bacillus diluent liquid preparation and liquid preparation storage method - Google Patents
Bacillus diluent, bacillus diluent liquid preparation and liquid preparation storage method Download PDFInfo
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- CN105695374A CN105695374A CN201610251912.5A CN201610251912A CN105695374A CN 105695374 A CN105695374 A CN 105695374A CN 201610251912 A CN201610251912 A CN 201610251912A CN 105695374 A CN105695374 A CN 105695374A
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- diluent
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- 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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- 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
- C12N1/04—Preserving or maintaining viable microorganisms
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Abstract
The invention discloses a bacillus diluent. The bacillus diluent is prepared from, by weight, 0.8% of sodium chloride, 5-50% of glycerol and the balance sterilization water. The PH value of the diluent is 8.6-10.6. The invention further discloses a bacillus diluent liquid preparation, and the concentration of bacillus diluent is 0.5*10<8>/ml. The invention further discloses a bacillus diluent liquid preparation storage method. The method comprises the steps of regulating diluted liquid with glycine and sodium hydroxide till PH becomes 10.0, and storing the liquid at normal temperature in the dark. By the adoption of the diluent, it can be guaranteed that spore count of bacilli in the liquid preparation and strain reactivation rate are high under corresponding storage conditions, the using effect in the clinical application process is guaranteed, and long-term effective storage of the bacillus diluent liquid preparation is made possible.
Description
Technical field
The invention belongs to microorganism field, be specifically related to the diluent of a kind of bacillus cereus liquid preparation, and this diluent dilutes the bacillus cereus liquid preparation obtained and the store method of bacillus cereus liquid preparation。
Background technology
Along with socioeconomic development, people are increasing to the demand of aquaculture products, and aquaculture progressively develops to the direction of industrialization, large-scale cultivation。The disabling additive such as highdensity aquaculture model, antibiotic abuse, olaquindox illegal uses also culture fishery to be developed in a healthy way and causes serious harm, and the negative effect such as residual toxin, pollution of ecological environment, threat health becomes the bottleneck of whole industry healthy development。In recent years, microbial ecological agent popularization and application in animal and veterinary, human health etc., great many of experiments and production practices confirm the application curative effect of microbial ecological agent, and its application in marine industry is also increasingly by the attention of people。Microbial ecological agent has the advantage such as green, environmental protection, and in cultivation, microbial ecological agent can be used to solve aquaculture and pollutes the problem with disease。Nature widely distributed bacillus cereus, multiple enzyme can be produced, be one of animals and plants microecosystem dominant population, play an important role in aquaculture。
Bacillus cereus is as additive used for aquiculture, and some researchs prove that it can promote breed variety acceleration of growth, reduce feed coefficient, maintains intestinal microecology balance, strengthens animal body immunity, improves resistance against diseases and reduces pollution etc.。
Compared with the natural water such as lake, river, artificial cultivation pond is the ecosystem of manual control, various physicochemical properties therein and the sufficiently complex fragility of biotic factor relation, and along with the carrying out of cultivation, water body environment is easily subject to the impact of each side factor。Microorganism in artificial cultivation pond is the important component part of its ecosystem, and microbe species in water body and content are easily subject to the impact of external environment condition。In breeding process, add the beneficial microbe of external source, it is possible to change the microbe species in water body and quantity with comparalive ease。By artificial regulatory, the microorganism composition of Optimal culture water body environment and quantity, promote to decompose the organic pollution such as remaining feedstuff and excretion refuse that produce in the breeding process of pond, it is suppressed that the growth and breeding of pathogenic microorganism。Bacillus cereus application in aquaculture has been carried out exploratory development by many aquaculture experts, and result display bacillus cereus has good denitrogenation dephosphorization activity, uses bacillus cereus can effectively reduce COD in water body (COD), NO in breeding water body2 ——N、NO3 ——N、NH4 +N and PO4 3—The concentration of P, improves breeding water body quality。
But, bacillus preparation application in aquaculture, also in primary developmental stage, lacks the theoretical direction of maturation in actual application, and many problems need to be studied further。On present fish market, the product of circulation mainly has liquid and pulverous bacillus cereus product, but the market product quality of the liquid formulation of bacillus cereus is unstable, do not have unified standard, application effect gap big, causes bacillus cereus liquid preparation popularization resistance in aquaculture industry bigger。
Summary of the invention
An object of the present invention is in that according to the above industry issue existed, it is provided that a kind of bacillus cereus diluent。
The technical solution adopted for the present invention to solve the technical problems is: a kind of bacillus cereus diluent, includes following component by weight percentage: the sodium chloride of 0.8%, and the glycerol of 5 50%, and the aquesterilisa of surplus, the pH value of diluent is 8.6 10.6。
Preferably, described a kind of bacillus cereus diluent, include following component by weight percentage: the sodium chloride of 0.8%, the glycerol of 15%, and the aquesterilisa of surplus, the pH value of diluent is 10.0。
The two of the purpose of the present invention are to provide the bacillus cereus liquid preparation adopting above-mentioned diluted to obtain。
The technical solution adopted for the present invention to solve the technical problems is: a kind of bacillus cereus liquid preparation, the concentration of bacillus cereus is 0.5 × 108/ml。
The three of the purpose of the present invention are to provide the store method of the bacillus cereus liquid preparation adopting above-mentioned diluted to obtain, and the method is able to maintain that the spore state of bacillus cereus and ensures the resurrection rate of strain。
The technical solution adopted for the present invention to solve the technical problems is: the store method of a kind of bacillus cereus liquid preparation, comprises the following steps:
Utilizing the liquid that glycine and sodium hydroxide regulate after diluting is 10.0 to pH value, is placed in room temperature, the preservation of backlight place, in order to using。
The invention has the beneficial effects as follows:
The present invention proposes the dilution suitable in relatively common fermentation of bacillus product and preservation, and there is good ferment effect, it is possible under simple preservation condition, bacillus cereus is carried out the preservation up to 9 months, maintain its bacillus cereus and be in spore state, improve the resurrection rate of bacillus cereus。
Diluent of the present invention and its store method have that cost is low, simple to operate, strain resurrection rate high, not only well maintain spore number and the resurrection rate of bacillus cereus liquid preparation, but also simplify operating procedure, reduce input cost, greatly facilitate the development of liquid microecological bacillus preparation。
Detailed description of the invention
Below in conjunction with embodiment, the present invention is described in further detail。
The invention discloses a kind of bacillus cereus diluent, include following component by weight percentage: the sodium chloride of 0.8%, the glycerol of 5 50%, and the aquesterilisa of surplus, the pH value of diluent is 8.6 10.6。
As preferred embodiment, including following component by weight percentage: the sodium chloride of 0.8%, the glycerol of 15%, and the aquesterilisa of surplus, the pH value of diluent is 10.0。
The invention also discloses a kind of bacillus cereus liquid preparation, successful for fermentation bacillus cereus product is collected, and bacillus cereus stock solution is carried out plate count, and then utilize aforementioned bacillus cereus diluent to be diluted obtaining to stock solution, the concentration of bacillus cereus is 0.5 × 108/ml。
The collection of fermentation of bacillus stock solution and counting
The bacillus cereus stock solution fermented is collected, processes, and fermenation raw liquid is carried out plate count, step: successively fermenation raw liquid is diluted 10 respectively—1、10—2、10—3、10—4、10—5、10—6、10—7 -----, bacterium colony is counted。
The dilution of bacillus cereus stock solution
Utilize 0.8%NaCl that bacillus cereus stock solution is diluted, be simultaneously introduced glycerol to final concentration respectively 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, bacillus cereus final concentration of 0.5 × 108Individual/ml, is then placed in room temperature, backlight place by the solution after dilution。Within every 2 weeks, utilize simple dyeing that Bacillus amount is observed, utilize plate count law technology simultaneously。
The invention also discloses the store method of the bacillus cereus liquid preparation that a kind of cost is low, simple to operate, spore resurrection rate is high, comprise the following steps: utilize glycine and sodium hydroxide to regulate the liquid pH value after diluting to 10, be placed in room temperature, backlight place preserves, bacillus cereus can be made to maintain the effect of spore state, and the holding time was up to 9 months。
PH condition after bacillus cereus dilution is groped
Liquid pH after utilizing glycine and sodium hydroxide to regulate dilution is respectively as follows: 8.6,9.0,9.4,9.8,10.0,10.4,10.6, is simultaneously introduced glycerol to final concentration of 15%, bacillus cereus final concentration of 0.5 × 108Individual/ml, utilizes simple dyeing that Bacillus amount is observed for every 2 weeks, utilizes plate count law technology simultaneously。
Bacillus cereus diluent in the present invention, liquid formulation and store method not only well maintain strain amount and the resurrection rate of bacillus cereus liquid preparation, and, simplify operating procedure, reduce input cost, appear on the market but without method of the present invention clinically at present。
From specific experiment data result, the diluent formula of the present invention and preservation condition, it is possible to realize under conventional storage conditions, maintain spore state be basically unchanged up to 9 months, this be bacillus cereus liquid preparation standardization preservation provide guarantee。
Above-described embodiment is illustrative principles of the invention and effect thereof only; and the embodiment that part is used, for the person of ordinary skill of the art, without departing from the concept of the premise of the invention; can also making some deformation and improvement, these broadly fall into protection scope of the present invention。
Claims (4)
1. a bacillus cereus diluent, it is characterised in that include following component by weight percentage: the sodium chloride of 0.8%, the glycerol of 5 50%, and the aquesterilisa of surplus, the pH value of diluent is 8.6 10.6。
2. a kind of bacillus cereus diluent according to claim 1, it is characterised in that including following component by weight percentage: the sodium chloride of 0.8%, the glycerol of 15%, and the aquesterilisa of surplus, the pH value of diluent is 10.0。
3. one kind adopts the bacillus cereus liquid preparation that diluted as claimed in claim 1 obtains, it is characterised in that: the concentration of bacillus cereus is 0.5 × 108/ml。
4. the store method of the bacillus cereus liquid preparation adopting diluted as claimed in claim 1 to obtain, it is characterised in that comprise the steps:
Utilizing the liquid that glycine and sodium hydroxide regulate after diluting is 10.0 to pH value, is placed in room temperature, the preservation of backlight place。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610251912.5A CN105695374A (en) | 2016-04-21 | 2016-04-21 | Bacillus diluent, bacillus diluent liquid preparation and liquid preparation storage method |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610251912.5A CN105695374A (en) | 2016-04-21 | 2016-04-21 | Bacillus diluent, bacillus diluent liquid preparation and liquid preparation storage method |
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| CN105695374A true CN105695374A (en) | 2016-06-22 |
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| CN201610251912.5A Pending CN105695374A (en) | 2016-04-21 | 2016-04-21 | Bacillus diluent, bacillus diluent liquid preparation and liquid preparation storage method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113200611A (en) * | 2020-11-05 | 2021-08-03 | 青岛诺安百特生物技术有限公司 | Composition for improving liquid microecological preparation bottle expansion and application thereof |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105385599A (en) * | 2016-01-21 | 2016-03-09 | 南京医科大学第一附属医院 | Coprophilous fungus preservation solution and method for preserving coprophilous fungi |
-
2016
- 2016-04-21 CN CN201610251912.5A patent/CN105695374A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105385599A (en) * | 2016-01-21 | 2016-03-09 | 南京医科大学第一附属医院 | Coprophilous fungus preservation solution and method for preserving coprophilous fungi |
Non-Patent Citations (3)
| Title |
|---|
| 彭孝斌: ""预防医学情报杂志"", 《预防医学情报杂志》 * |
| 方光远等: ""几种不同条件下细菌菌种保存试验"", 《金陵科技学院学报》 * |
| 钟志宏等: ""甘油保存菌种方法的改良"", 《实用预防医学》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113200611A (en) * | 2020-11-05 | 2021-08-03 | 青岛诺安百特生物技术有限公司 | Composition for improving liquid microecological preparation bottle expansion and application thereof |
| CN113200611B (en) * | 2020-11-05 | 2023-02-03 | 青岛诺安百特生物技术有限公司 | Composition for improving liquid microecological preparation bottle expansion and application thereof |
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