CN112940990A - Preparation method and application of microbial agent activated and expanded by nano bubbles - Google Patents
Preparation method and application of microbial agent activated and expanded by nano bubbles Download PDFInfo
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- CN112940990A CN112940990A CN202110411548.5A CN202110411548A CN112940990A CN 112940990 A CN112940990 A CN 112940990A CN 202110411548 A CN202110411548 A CN 202110411548A CN 112940990 A CN112940990 A CN 112940990A
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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
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/50—Treatments combining two or more different biological or biochemical treatments, e.g. anaerobic and aerobic treatment or vermicomposting and aerobic treatment
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/80—Soil conditioners
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G5/00—Fertilisers characterised by their form
- C05G5/20—Liquid fertilisers
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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
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Abstract
The invention belongs to the technical field of preparation methods of microbial agents, in particular to a preparation method and application of a microbial agent activated and expanded by nano bubbles, which solves the problems that the number of colonies is limited and the effective viable bacteria rate is limited after the expansion culture of EM stock seeds in the prior art; the preparation method of the microbial agent activated by nano bubbles and expanded cultured comprises the following steps: adding sterile brown sugar water, honey and strain liquid into a primary fermentation tank together, and fermenting by combining an expanding culture process and a micro-nano bubble generation device; and (3) filling the activated strain liquid, sterile water and sterile brown sugar water into a secondary fermentation tank together, and fermenting by combining an expanding culture process and using a micro-nano bubble generating device. The invention greatly improves the effective viable bacteria rate of the EM original composite bacterial colony group, has higher purity and reduces the mixed bacteria rate to be below 0.1 percent.
Description
Technical Field
The invention relates to the technical field of preparation methods of microbial agents, in particular to a preparation method and application of a microbial agent activated and expanded by nano bubbles.
Background
EM is an abbreviation of English (Effective Microorganisms), and Chinese translates into: an "effective microbiota". It is composed of more than 80 microorganisms of 5 families, 10 genera, such as photosynthetic bacteria, lactic acid bacteria flora, yeast flora, actinomycete flora, filamentous flora, and the like, and is successfully researched by professor of Botiaea caraghua of university of Youki, Japan, and the like, which are famous applied microbiology in the world. The EM technology is a bioengineering technology with the largest application range in the world at present, and compared with a common biological preparation, the EM technology has the advantages of complex structure, stable performance and complete functions, and shows unprecedented high-tech level. According to the related data report, EM is widely applied to various fields of planting industry, breeding industry, environmental protection, human body health care and the like.
Due to the wide range of EM functions, the EM has strong advantages in the market, and various EM technical products in the market are produced at present. However, the number of colonies after the traditional EM stock culture is expanded is limited, and the effective viable bacteria rate is limited; and the biological activity has no obvious advantages compared with the original microorganism system in the nature, and the microbial agent is easy to be dissimilatory and has high mixed bacteria rate after being applied and put. Based on the statement, the invention provides a preparation method and application of a microbial agent activated and expanded by nano bubbles.
Disclosure of Invention
The invention aims to solve the problems that the number of colonies is limited after EM stock is expanded and cultured and the effective viable bacteria rate is limited in the prior art; and the biological activity has no obvious advantages compared with the original microbial system in the nature, and the problems of easy metamorphosis and high mixed bacteria rate after application and delivery are solved.
The preparation method of the microbial agent activated by nano bubbles and expanded culture comprises the following steps:
s1, strain activation:
s11, fermentation tank and auxiliary tool processing procedure;
s12, adding sterile brown sugar water, honey and strain liquid into the primary fermentation tank together, stirring uniformly, performing sealed fermentation by combining an expanding culture process and a micro-nano bubble generation device, and activating the strain liquid together;
s2, preparing microbial agent bacterial liquid:
s21, filling the activated strain liquid and sterile water into a secondary fermentation tank, adding sterile brown sugar water, uniformly stirring, sealing the fermentation tank, and performing anaerobic fermentation for 7-10 days by combining an expanding culture process and a micro-nano bubble generation device;
s22, after fermentation is completed, filling the fermentation finished product into a finished product barrel to obtain the required microbial inoculum;
s3, post-treatment process:
the fermentor was washed and the wash was collected in a closed loop with water.
Preferably, the fermentation tank and auxiliary equipment treating step in step S11 is specifically performed as follows: cleaning the needed fermentation tank and auxiliary appliances, sterilizing by ultraviolet rays, heating the fermentation tank with water for one day, and keeping the heating temperature at 35 ℃.
Preferably, the sterile brown water in steps S12 and S21 is prepared by the following method: adding brown sugar into 100 deg.C boiled water, decocting for 2-3min, dissolving brown sugar, and cooling to 35 deg.C.
Preferably, the temperature of the expanding culture process in the steps S12 and S21 is kept at 35 ℃.
The invention also provides an application method of the microbial agent activated and spread-cultured by adopting the nano bubbles, which is a three-linkage application method formed by a micro-nano bubble generating device, the microbial agent and a liquid organic fertilizer.
Preferably, the liquid organic fertilizer is a humic acid-containing water-soluble fertilizer.
The preparation method and the application of the microbial agent activated and expanded by the nano bubbles have the following beneficial effects:
1. the microbial agent provided by the invention is based on an EM (effective microorganism) stock composite bacterial colony group, a nano-micron gas-liquid interface technology is organically integrated into the whole process of the EM stock composite bacterial colony group expansion culture, namely in the process links of activating strains and expanding culture, a micro-nano bubble generating device is used for injecting nano bubbles, so that the effective viable bacteria rate of the EM stock composite bacterial colony group is greatly improved, the purity is higher, the mixed bacteria rate is reduced to be below 0.1 percent (the national standard specification is less than 10 percent, namely the product is qualified), and the harmless index reaches and is far superior to the national standard (GB 20287-2006).
2. The microbial agent produced by the method through the nano-micron gas-liquid interface technology is in a liquid state, is dark brown in color, and has sour taste and wine aroma and sweet taste; the microbial agent prepared by the invention has the colony number more than 6 times higher than that of the EM bacteria sold in the market at present, the safety of seed culture is fully verified, and the microbial agent is suitable for organic fertilizer fermentation production, agricultural seed culture, soil improvement, tail water treatment of seed culture, water environment treatment, sewage treatment and the like; the application process of the items is combined with a nano-micron gas-liquid interface technology, so that the use effect of the microbial agent can be optimal.
3. The invention also provides an application method of the microbial agent activated and expanded by the nano bubbles, namely a three-linkage application method consisting of the micro-nano bubble generating device, the microbial agent and a liquid organic fertilizer (containing humic acid water-soluble fertilizer), which is applied to the fields of agricultural seed culture, soil improvement, seed culture tail water treatment, water environment treatment and sewage treatment, has been practiced for many years, has safe and reliable effect and is worthy of popularization and application.
4. The micro-nano bubble generating device generates micro-bubbles in a physical mode, and in the implementation process of the triple linkage application method:
physical action: mainly shows the air flotation effect generated by micron bubbles, and has the effects of liquid-liquid separation and liquid-solid separation, deconstruction of long-term retention pollutants in water molecular groups and separation of pesticide residue antibiotics;
chemical action: the quasi-nano bubbles exist in water for several days, so that the three-nitrogen conversion in an underwater anaerobic environment is greatly promoted, a series of effects such as the oxidation-reduction potential of a water body are improved, and the effective utilization rate of a liquid organic fertilizer (containing humic acid water-soluble fertilizer) is improved;
biological effect: the nano bubbles can be stored under water for more than several weeks, so that the activity of the microbial agent is greatly improved and maintained, and the on-line in-situ and muddy water co-treatment is realized by indirectly strengthening the nitrogen fixation function of microorganisms, improving the bottom sediment of the substrate, preventing the eutrophication of the water body, repairing the self-purification capacity of the breeding water body and the like.
Drawings
FIG. 1 is a process flow diagram of a method for preparing a microbial inoculum by nano-bubble activation and propagation according to the present invention;
FIG. 2 is a report of detection of beneficial bacterial communities by using nanobubble activated and expanded microbial agents according to an embodiment of the present invention;
FIG. 3 is a safety detection report of a microbial inoculum activated and expanded by nanobubbles according to an embodiment of the present invention.
Detailed Description
The present invention will be further illustrated with reference to the following specific examples.
Example one
The invention provides a preparation method of a microbial agent activated and expanded by nano bubbles, which comprises the following steps:
s1, strain activation:
s11, fermentation tank and auxiliary appliance treatment process, wherein the specific operation is that after the required fermentation tank and auxiliary appliance are cleaned and sterilized by ultraviolet rays, the fermentation tank is heated by water feeding for one day, and the heating temperature is kept at 35 ℃;
s12, adding brown sugar into boiled water of 100 ℃ for boiling for 2min, cooling to 35 ℃ after dissolving the brown sugar to obtain sterile brown sugar water, adding the sterile brown sugar water, honey and strain liquid into a primary fermentation tank together, stirring uniformly, performing sealed fermentation by combining a spread culture process and a micro-nano bubble generation device, keeping the temperature at 35 ℃ in the spread culture process, and activating the strain liquid together;
s2, preparing microbial agent bacterial liquid:
s21, filling the activated strain liquid and sterile water into a secondary fermentation tank, adding sterile brown sugar water, uniformly stirring, sealing the fermentation tank, performing anaerobic fermentation for 7 days by combining an expanding culture process and a micro-nano bubble generation device, and keeping the temperature in the expanding culture process at 35 ℃;
s22, after fermentation is completed, filling the fermentation finished product into a finished product barrel to obtain the required microbial inoculum;
s3, post-treatment process:
the fermentor was washed and the wash was collected in a closed loop with water.
Example two
The invention provides a preparation method of a microbial agent activated and expanded by nano bubbles, which comprises the following steps:
s1, strain activation:
s11, fermentation tank and auxiliary appliance treatment process, wherein the specific operation is that after the required fermentation tank and auxiliary appliance are cleaned and sterilized by ultraviolet rays, the fermentation tank is heated by water feeding for one day, and the heating temperature is kept at 35 ℃;
s12, adding brown sugar into boiled water of 100 ℃ for boiling for 3min, cooling to 35 ℃ after dissolving the brown sugar to obtain sterile brown sugar water, adding the sterile brown sugar water, honey and strain liquid into a primary fermentation tank together, stirring uniformly, performing sealed fermentation by combining a spread culture process and a micro-nano bubble generation device, keeping the temperature at 35 ℃ in the spread culture process, and activating the strain liquid together;
s2, preparing microbial agent bacterial liquid:
s21, filling the activated strain liquid and sterile water into a secondary fermentation tank, adding sterile brown sugar water, uniformly stirring, sealing the fermentation tank, performing anaerobic fermentation for 8 days by combining an expanding culture process and a micro-nano bubble generation device, and keeping the temperature in the expanding culture process at 35 ℃;
s22, after fermentation is completed, filling the fermentation finished product into a finished product barrel to obtain the required microbial inoculum;
s3, post-treatment process:
the fermentor was washed and the wash was collected in a closed loop with water.
EXAMPLE III
The invention provides a preparation method of a microbial agent activated and expanded by nano bubbles, which comprises the following steps:
s1, strain activation:
s11, fermentation tank and auxiliary appliance treatment process, wherein the specific operation is that after the required fermentation tank and auxiliary appliance are cleaned and sterilized by ultraviolet rays, the fermentation tank is heated by water feeding for one day, and the heating temperature is kept at 35 ℃;
s12, adding brown sugar into boiled water of 100 ℃ for boiling for 2min, cooling to 35 ℃ after dissolving the brown sugar to obtain sterile brown sugar water, adding the sterile brown sugar water, honey and strain liquid into a primary fermentation tank together, stirring uniformly, performing sealed fermentation by combining a spread culture process and a micro-nano bubble generation device, keeping the temperature at 35 ℃ in the spread culture process, and activating the strain liquid together;
s2, preparing microbial agent bacterial liquid:
s21, filling the activated strain liquid and sterile water into a secondary fermentation tank, adding sterile brown sugar water, uniformly stirring, sealing the fermentation tank, performing anaerobic fermentation for 10 days by combining an expanding culture process and a micro-nano bubble generation device, and keeping the temperature in the expanding culture process at 35 ℃;
s22, after fermentation is completed, filling the fermentation finished product into a finished product barrel to obtain the required microbial inoculum;
s3, post-treatment process:
the fermentor was washed and the wash was collected in a closed loop with water.
The microbial agent prepared in the first embodiment of the invention is detected according to the national mandatory standard GB20287-2006 agricultural microbial agent, and the obtained result is as follows:
the colony numbers of the microbial inoculum prepared in the first embodiment of the invention and the commercial EM bacteria in different culture media are compared and tested, and the following results are obtained:
| number of commercially available EM bacterial colonies | EXAMPLE one microbial inoculum colony count | |
| Honey sugar water culture medium | 4.97×106 | 3.29×107 |
| Broth culture medium | 4.23×105 | 2.67×106 |
| Nutrient agar culture medium | 1.96×105 | 1.17×106 |
The invention also provides an application method of the microbial agent activated and spread-cultured by adopting the nano bubbles, which is a three-linkage application method formed by a micro-nano bubble generating device, the microbial agent and a liquid organic fertilizer, and the application method specifically comprises the following operations:
the microbial agent provided by the invention has been practiced for many years when applied to agricultural breeding, has safe and reliable effect, and can provide a general scheme:
the technical scheme of 'three-linkage' in aquaculture or ecological mixed culture of rice is as follows:
1. every season of aquaculture (early summer to winter), conventional farmers typically invest in the following biologicals: the fertilizer is a water-fertilizing paste, 100 (calcium) is quickly supplemented, an antidote, EM water-adjusting king, nitrite-adjusting medicine, brown sugar, bottom cover medicine and the like, and the total cost is about 1000 yuan/mu.
2. The microbial agent cultured by adopting the nano-micro gas-liquid interface technology, the liquid organic fertilizer (containing humic acid water-soluble fertilizer) and the micro-nano bubble generating device matched with the nano-micro gas-liquid interface technology in the using process of the bacteria/fertilizer are called as the 'triple linkage' technology of the nano-micro gas-liquid interface technology in agricultural breeding application.
3. The use of the microbial inoculum: about 1 jin per mu per week. The microbial inoculum can be directly put into the aquaculture water body (sprayed after dilution to ensure good dispersion), and the microbial inoculum diluted by 200 times is recommended to be added into the feed. When the microbial inoculum is directly fed or mixed, the microbial inoculum is prevented from being fed in 2 meters of the nozzles of the aerator and the micro-nano bubble generating device; after the microbial inoculum is directly fed or mixed and fed, the aerator and the micro-nano bubble generating device are recommended to be started up for 1 hour or more.
Description of the drawings: the food is kept wet, and the food is fed after waking for half an hour. If 5 jin of water should be put in the foodstuff and the ideal humidity can be reached (the fist is released after grasping and the state is loose), then the microbial inoculum put in 5 jin of water is: 5 jin (2500 g) ÷ 200 times =12.5 g (bacterial agent)
4. Liquid fertilizer: the academy of the water-soluble fertilizer (organic fertilizer) containing humic acid is that the fertilizer is only required to be put in every season when adult fish is cultured (the putting in of fry/shrimp fry/crab fry and the like is recommended to be finished 4-5 times in 1 month), and about 20 jin =300 yuan/mu. The throwing mode is uniform and direct throwing, and after throwing, the aerator and the micro-nano bubble generating device are recommended to be started for 1 hour or more. As the culture water body becomes thin after the microbial inoculum is used, the organic liquid fertilizer is an important supplement.
5. Under the conventional condition, the micro-nano bubble generating device recommends to start up at night, and recommends to start up for 1 hour when adding the microbial inoculum and the liquid fertilizer.
6. By adopting the 'triple linkage' technology, the investment of other biological agents for farmers can be greatly reduced, and the total cost is 160 (microbial inoculum) +300 (liquid fertilizer) +200 or so (other) = about 660 yuan/mu/season.
7. Pre-treatment of the breeding season:
a. one week before water is injected into the fishpond or the paddy field, the microbial inoculum diluted by 200 times is uniformly sprayed, and the amount of the microbial inoculum diluted by 1 jin per square meter.
b. After water injection and 3-10 days before stocking, the microbial agent is sprayed on the water surface, one mu of pond is used for 2 jin according to the depth of one meter, and the usage amount is properly increased in places with poor water quality. The process is matched with the micro-nano bubble generating device for use, and the effect is better.
The microbial agent can promote the activation and growth of normal flora and beneficial algae in a culture ecological system and keep the ecological balance of culture water; the bait is mixed for feeding, so that the absorption function and the disease and stress resistance of the fish are directly enhanced, and the robust growth is promoted; in addition, the photosynthetic bacteria in the microbial agent stock solution can remove the ammonium nitrite in the water by utilizing the denitrification of the hydrogen sulfide, the organic acid, the ammonia and the amino acid in the water, thereby purifying the excrement and the residual bait in the culture pond, improving the water quality and reducing the diseases of fishes and shrimps.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (6)
1. The preparation method of the microbial agent activated by nano bubbles and expanded culture is characterized by comprising the following steps:
s1, strain activation:
s11, fermentation tank and auxiliary tool processing procedure;
s12, adding sterile brown sugar water, honey and strain liquid into the primary fermentation tank together, stirring uniformly, performing sealed fermentation by combining an expanding culture process and a micro-nano bubble generation device, and activating the strain liquid together;
s2, preparing microbial agent bacterial liquid:
s21, filling the activated strain liquid and sterile water into a secondary fermentation tank, adding sterile brown sugar water, uniformly stirring, sealing the fermentation tank, and performing anaerobic fermentation for 7-10 days by combining an expanding culture process and a micro-nano bubble generation device;
s22, after fermentation is completed, filling the fermentation finished product into a finished product barrel to obtain the required microbial inoculum;
s3, post-treatment process:
the fermentor was washed and the wash was collected in a closed loop with water.
2. The method of claim 1, wherein the step S11 of processing the fermenter and the auxiliary tool comprises the following steps: cleaning the needed fermentation tank and auxiliary appliances, sterilizing by ultraviolet rays, heating the fermentation tank with water for one day, and keeping the heating temperature at 35 ℃.
3. The method for preparing microbial inoculant by nanobubble activation and propagation according to claim 1, wherein the sterile brown sugar water of steps S12 and S21 is prepared by the following steps: adding brown sugar into 100 deg.C boiled water, decocting for 2-3min, dissolving brown sugar, and cooling to 35 deg.C.
4. The method of claim 1, wherein the steps S12 and S21 are performed in a step S12.
5. An application method of the microbial inoculum activated and spread cultured by the nanobubbles according to any one of claims 1 to 4, which is characterized in that the application method is a 'three-linkage' application method which is composed of a nanobubble generating device, the microbial inoculum and a liquid organic fertilizer.
6. The method for applying the microbial inoculant activated by nanobubbles and spread cultured according to claim 5, wherein the liquid organic fertilizer is a humic acid-containing water-soluble fertilizer.
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| CN114875078A (en) * | 2022-06-16 | 2022-08-09 | 东华大学 | Process for preparing short chain fatty acids |
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| CN114875078B (en) * | 2022-06-16 | 2023-11-17 | 东华大学 | Process for preparing short-chain fatty acids |
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