CN110938576A - Preparation method of efficient spore flora for decontamination of aquaculture - Google Patents
Preparation method of efficient spore flora for decontamination of aquaculture Download PDFInfo
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- CN110938576A CN110938576A CN201911386127.0A CN201911386127A CN110938576A CN 110938576 A CN110938576 A CN 110938576A CN 201911386127 A CN201911386127 A CN 201911386127A CN 110938576 A CN110938576 A CN 110938576A
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Abstract
The invention discloses a method for preparing efficient spore flora for decontamination of aquaculture, which solves the problems that the prior spore flora in an aquaculture pond has less dominant strains, so that the spore flora can not efficiently and quickly explain macromolecular organic matters in water in the aquaculture pond, thereby influencing the water quality of the aquaculture pond and further causing the cultured aquatic organisms to be incapable of healthy and quick growth, separates and preferentially cultivates the spore flora with comprehensive super-strong degradation function and ultra-low oxygen consumption from pond mud by screening the indigenous dominant spore strains and adopting the advanced microbial engineering technology, then cultures, optimizes and compounds the spore flora to ensure that the strains are pure, thereby regulating and protecting the water quality in the aquaculture pond by using a small amount of spore flora when the aquaculture pond is decontaminated and degraded, thereby ensuring that the water quality in the aquaculture pond can lead the aquatic organisms to grow healthily and rapidly.
Description
Technical Field
The invention relates to the technical field of preparation methods of spore flora, in particular to a preparation method of efficient spore flora for decontamination of aquaculture.
Background
The sludge at the bottom of the aquaculture pond contains a large amount of spore bacteria, and the spore bacteria can accelerate the decomposition of macromolecular organic matters in the aquaculture pond, so that the water quality in the aquaculture pond can be conveniently regulated and protected, and the healthy and rapid growth of organisms in the aquaculture pond can be ensured; however, the spore bacteria in the existing aquaculture pond have the following problems: the dominant bacteria in the spore bacteria in the existing aquaculture pond are less, so that the spore bacteria can not efficiently and quickly explain macromolecular organic matters in the water body of the aquaculture pond, the water quality of the aquaculture pond is influenced, and cultured aquatic organisms can not grow healthily and quickly.
Disclosure of Invention
The invention aims to provide a preparation method of a high-efficiency spore flora for decontamination of aquaculture, aiming at solving the problem.
The invention is realized by the following steps:
a high-efficiency spore flora culture optimization method for aquaculture decontamination comprises the following steps:
s1: sample collection
Collecting sludge at the bottom of a 5-15 cm aquaculture pond, putting the sludge into a 50ml sterilization centrifugal tube, and then carrying the sludge back to a laboratory for separation;
s2: preparation of enrichment Medium
The raw materials for preparing the enrichment medium are as follows: ultrapure water, peptone, yeast extract, starch, MgSO4, KH2PO4 and Na2HPO 4;
dissolving peptone, yeast extract, starch, MgSO4, KH2PO4 and Na2HPO4 in ultrapure water by weight, adjusting the pH of the solution to 7.8, and sterilizing at 121 deg.C for 20 min;
s3: preparation of isolation Medium
The raw materials for preparing the separation culture medium are as follows: ultrapure water, MnSO4, glucose, agar, peptone, yeast extract and KH2PO 4;
dissolving glucose, agar, peptone, yeast extract and KH2PO4 in ultrapure water by weight, heating and boiling the solution, adding 1ml of 3.08% MnSO4, and sterilizing the solution at 121 deg.C for 30 min;
s4: bacterial colony culture
Inoculating the collected sample into an enrichment medium according to the amount of 1/5, culturing for 24 hours at 30 ℃, placing the cultured bacterial suspension into a water bath kettle at 80 ℃ for heating for 10-20 minutes to kill other bacteria which can not form spores, then diluting the treated bacterial suspension by 10-fold gradient, absorbing diluents with different concentrations, inoculating the bacterial suspension into a spore isolation medium by a plate coating method, culturing for 24-48 hours at 30 ℃, picking suspected bacterial colonies, firstly carrying out simple dyeing observation, picking bacillus bacterial colonies for pure culture, and storing on an inclined plane.
Further, the preparation raw material of the enrichment medium contained 1000ml of ultrapure water, 10g of peptone, 3g of yeast extract, 3g of starch, 0.1g of MgSO4, 1.5g of KH2PO4, and 2g of Na2HPO 4.
Further, the isolation medium was prepared from 1000ml of ultrapure water, 1ml of MnSO4, 5g of glucose, 20g of agar, 5g of peptone, 4g of yeast extract, and 5g of KH2PO 4.
A preparation process flow of efficient spore flora for decontamination of aquaculture comprises the following steps:
s1: preparation of bacillus subtilis liquid
Placing the bacillus subtilis strain in a liquid culture medium for culturing to prepare bacillus subtilis liquid, and then heating a proper amount of brown sugar in the bacillus subtilis liquid to propagate the bacillus subtilis;
s2: fermentation of
Carrying out temperature and humidity control on the bacillus subtilis liquid, mixing the bacillus subtilis liquid with the jade bran, carrying out temperature control culture and fermenting;
s3: drying and packaging
And drying the mixture of the fermented jade bran and the bacillus subtilis liquid, weighing and bagging the dried product after drying, and sealing the product after bagging to finish the packaging of the finished product.
Furthermore, the mixture of the jade bran and the bacillus subtilis liquid is stirred and fermented in a mixer during fermentation, and the temperature-controlled culture is carried out on the mixture of the jade bran and the bacillus subtilis liquid in the fermentation process so as to ensure that the bacillus subtilis can be rapidly propagated.
Compared with the prior art, the invention has the beneficial effects that: according to the invention, through screening indigenous dominant spore strains, an advanced microbial engineering technology is adopted, spore flora with comprehensive super-strong degradation function and ultralow oxygen consumption is separated and preferably cultured from pond mud, and then the spore flora is cultured, optimized and compounded, so that the strains are pure, and therefore, a small amount of spore flora can be used for regulating and protecting the water quality in the aquaculture pond when the aquaculture pond is subjected to decontamination and degradation, so that the water quality in the aquaculture pond can be ensured to enable aquatic organisms to grow healthily and rapidly, and meanwhile, the spore flora has an obvious effect on decomposing macromolecular organic matters in aquaculture water.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are required to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
FIG. 1 is a flow chart of the preparation process of the efficient spore flora of the present invention;
FIG. 2 is a comparison chart of experimental data of a highly effective spore flora of the present invention;
FIG. 3 is a second comparison chart of experimental data of the highly effective spore flora of the present invention;
FIG. 4 is a third comparison chart of experimental data of the highly effective spore flora of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings of the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention. Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.
Example (b): referring to fig. 1, 2, 3 and 4: a high-efficiency spore flora culture optimization method for aquaculture decontamination comprises the following steps:
s1: sample collection
Collecting sludge at the bottom of a 5-15 cm aquaculture pond, putting the sludge into a 50ml sterilization centrifugal tube, and then carrying the sludge back to a laboratory for separation;
s2: preparation of enrichment Medium
The raw materials for preparing the enrichment medium are as follows: ultrapure water, peptone, yeast extract, starch, MgSO4, KH2PO4 and Na2HPO 4;
dissolving 10g of peptone, 3g of yeast extract, 3g of starch, 0.1g of MgSO4, 1.5g of KH2PO4 and 2g of Na2HPO4 in 1000ml of ultrapure water by weight, adjusting the pH of the solution to 7.8, and then sterilizing at 121 ℃ for 20 min;
s3: preparation of isolation Medium
The raw materials for preparing the separation culture medium are as follows: ultrapure water, MnSO4, glucose, agar, peptone, yeast extract and KH2PO 4;
dissolving 5g of glucose, 20g of agar, 5g of peptone, 4g of yeast extract and 5g of KH2PO4 in 1000ml of ultrapure water by weight, heating and boiling the dissolved solution, adding 1ml of 3.08% MnSO4, and sterilizing the dissolved solution at 121 ℃ for 30 min;
s4: bacterial colony culture
Inoculating the collected sample into an enrichment medium according to the amount of 1/5, culturing for 24 hours at 30 ℃, placing the cultured bacterial suspension into a water bath kettle at 80 ℃ for heating for 10-20 minutes to kill other bacteria which can not form spores, then diluting the treated bacterial suspension by 10-fold gradient, absorbing diluents with different concentrations, inoculating the bacterial suspension into a spore isolation medium by a plate coating method, culturing for 24-48 hours at 30 ℃, picking suspected bacterial colonies, firstly carrying out simple dyeing observation, picking bacillus bacterial colonies for pure culture, and storing on an inclined plane.
A preparation process flow of efficient spore flora for decontamination of aquaculture comprises the following steps:
s1: preparation of bacillus subtilis liquid
Placing the bacillus subtilis strain in a liquid culture medium for culturing to prepare bacillus subtilis liquid, and then heating a proper amount of brown sugar in the bacillus subtilis liquid to propagate the bacillus subtilis;
s2: fermentation of
The method comprises the following steps of carrying out temperature and humidity control on bacillus subtilis liquid, mixing the bacillus subtilis liquid with jade bran together, carrying out temperature control culture and fermentation, putting the jade bran and the bacillus subtilis liquid into a mixer during fermentation, stirring and fermenting, and carrying out temperature control culture on the mixture of the jade bran and the bacillus subtilis liquid during fermentation so as to ensure that the bacillus subtilis can be rapidly propagated;
s3: drying and packaging
And drying the mixture of the fermented jade bran and the bacillus subtilis liquid, weighing and bagging the dried product after drying, and sealing the product after bagging to finish the packaging of the finished product.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (5)
1. An efficient spore flora culture optimization method for aquaculture decontamination is characterized in that: the method comprises the following steps:
s1: sample collection
Collecting sludge at the bottom of a 5-15 cm aquaculture pond, putting the sludge into a 50ml sterilization centrifugal tube, and then carrying the sludge back to a laboratory for separation;
s2: preparation of enrichment Medium
The raw materials for preparing the enrichment medium are as follows: ultrapure water, peptone, yeast extract, starch, MgSO4, KH2PO4 and Na2HPO 4;
dissolving peptone, yeast extract, starch, MgSO4, KH2PO4 and Na2HPO4 in ultrapure water by weight, adjusting the pH of the solution to 7.8, and sterilizing at 121 deg.C for 20 min;
s3: preparation of isolation Medium
The raw materials for preparing the separation culture medium are as follows: ultrapure water, MnSO4, glucose, agar, peptone, yeast extract and KH2PO 4;
dissolving glucose, agar, peptone, yeast extract and KH2PO4 in ultrapure water by weight, heating and boiling the solution, adding 1ml of 3.08% MnSO4, and sterilizing the solution at 121 deg.C for 30 min;
s4: bacterial colony culture
Inoculating the collected sample into an enrichment medium according to the amount of 1/5, culturing for 24 hours at 30 ℃, placing the cultured bacterial suspension into a water bath kettle at 80 ℃ for heating for 10-20 minutes to kill other bacteria which can not form spores, then diluting the treated bacterial suspension by 10-fold gradient, absorbing diluents with different concentrations, inoculating the bacterial suspension into a spore isolation medium by a plate coating method, culturing for 24-48 hours at 30 ℃, picking suspected bacterial colonies, firstly carrying out simple dyeing observation, picking bacillus bacterial colonies for pure culture, and storing on an inclined plane.
2. The method for optimizing the culture of the efficient spore flora for decontaminating aquaculture of claim 1, wherein the preparation raw material of the enrichment medium comprises 1000ml of ultrapure water, 10g of peptone, 3g of yeast extract, 3g of starch, 0.1g of MgSO4, 1.5g of KH2PO4 and 2g of Na2HPO 4.
3. The method for optimizing the culture of the efficient spore flora for decontaminating aquaculture of claim 2, wherein the raw material prepared by the separation medium comprises 1000ml of ultrapure water, 1ml of MnSO4, 5g of glucose, 20g of agar, 5g of peptone, 4g of yeast extract and 5g of KH2PO 4.
4. The process of claim 3, wherein the process comprises the following steps:
s1: preparation of bacillus subtilis liquid
Placing the bacillus subtilis strain in a liquid culture medium for culturing to prepare bacillus subtilis liquid, and then heating a proper amount of brown sugar in the bacillus subtilis liquid to propagate the bacillus subtilis;
s2: fermentation of
Carrying out temperature and humidity control on the bacillus subtilis liquid, mixing the bacillus subtilis liquid with the jade bran, carrying out temperature control culture and fermenting;
s3: drying and packaging
And drying the mixture of the fermented jade bran and the bacillus subtilis liquid, weighing and bagging the dried product after drying, and sealing the product after bagging to finish the packaging of the finished product.
5. The process flow of claim 3, wherein the mixture of corn bran and the bacillus subtilis solution is stirred and fermented in a mixer during fermentation, and the temperature-controlled culture is performed on the mixture of corn bran and the bacillus subtilis solution during fermentation to ensure the bacillus subtilis can be rapidly propagated.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586156A (en) * | 2012-03-10 | 2012-07-18 | 安徽联大生物环保科技有限公司 | Method for simply and rapidly separating aerobic denitrification bacillus |
| CN102618474A (en) * | 2012-04-10 | 2012-08-01 | 江苏今世缘酒业股份有限公司 | Bacillus subtilis and separate culture method for same |
| CN103875890A (en) * | 2012-12-19 | 2014-06-25 | 西安格润牧业发展有限公司 | Microbe forage and preparation method thereof |
| CN107043725A (en) * | 2017-04-25 | 2017-08-15 | 西安鑫汉宝生物科技有限公司 | Method of bacillus subtilis and bacillus coagulans mixed fermentation and application thereof |
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- 2019-12-29 CN CN201911386127.0A patent/CN110938576A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102586156A (en) * | 2012-03-10 | 2012-07-18 | 安徽联大生物环保科技有限公司 | Method for simply and rapidly separating aerobic denitrification bacillus |
| CN102618474A (en) * | 2012-04-10 | 2012-08-01 | 江苏今世缘酒业股份有限公司 | Bacillus subtilis and separate culture method for same |
| CN103875890A (en) * | 2012-12-19 | 2014-06-25 | 西安格润牧业发展有限公司 | Microbe forage and preparation method thereof |
| CN107043725A (en) * | 2017-04-25 | 2017-08-15 | 西安鑫汉宝生物科技有限公司 | Method of bacillus subtilis and bacillus coagulans mixed fermentation and application thereof |
Non-Patent Citations (1)
| Title |
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| 石俭 等: "高效产芽孢除磷菌株的筛选鉴定及影响其除磷的因素" * |
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