CN114133025A - Preparation method of microbial agent carrier material and application of microbial agent carrier material in high-density aquaculture - Google Patents
Preparation method of microbial agent carrier material and application of microbial agent carrier material in high-density aquaculture Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/02—Aerobic processes
- C02F3/10—Packings; Fillings; Grids
- C02F3/105—Characterized by the chemical composition
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F2003/001—Biological treatment of water, waste water, or sewage using granular carriers or supports for the microorganisms
- C02F2003/003—Biological treatment of water, waste water, or sewage using granular carriers or supports for the microorganisms using activated carbon or the like
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/105—Phosphorus compounds
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2209/00—Controlling or monitoring parameters in water treatment
- C02F2209/02—Temperature
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- C02F2209/06—Controlling or monitoring parameters in water treatment pH
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- C02F2209/14—NH3-N
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
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Abstract
A preparation method of a microbial agent carrier material and application in high-density aquaculture belong to the field of microbial technology and application, the microbial agent carrier material is prepared by five steps of bamboo charcoal pretreatment, culture medium preparation, bacterial liquid preparation, bacterial mud preparation, microorganism immobilization and sodium alginate wrapping, and is matched with a slow-release oxygen-increasing material to be applied to high-density aquaculture; can effectively regulate the stable structure of the intestinal microflora of the fishes, inhibit the growth of mixed bacteria, improve the utilization rate of bait, reduce nitrite and the like, and solve the problems of poor water quality stability, poor quality and safety of aquatic products and difficult improvement of culture density and unit benefit.
Description
Technical Field
The invention relates to the field of microbial technology and application, in particular to a preparation method of a microbial agent carrier material and application of the microbial agent carrier material in high-density aquaculture.
Background
With the rapid development and intensive operation of the aquaculture industry, a number of problems still exist:
(1) in conventional aquaculture in the current market, the content of ammonia nitrogen and nitrite is too high, the pH value is not proper, the Chemical Oxygen Demand (COD) is too high, the content of dissolved oxygen is too low, in a high-density aquaculture system, the deposition amount of organic pollutants such as residual baits, excrement, animal and plant carcasses and the like is large, the residual amount of medicine, excessive exogenous pollutants and the like are added, and the organic matters generate a large amount of harmful substances such as ammonia nitrogen, nitrite, hydrogen sulfide and the like under the decomposition action of anaerobic microorganisms, so that aquaculture animals are directly damaged;
(2) for the abnormality, the conventional treatment means is to continuously add medicines, antibiotics, chemical reagents and the like, and although the addition of the medicines can solve the problems transiently, the problems of too high content of ammonia nitrogen and nitrite, uncomfortable pH value, too high Chemical Oxygen Demand (COD), too low content of dissolved oxygen and the like cannot be fundamentally solved. But also introduces a large amount of toxic and harmful substances, which seriously harms the safety and health of food;
(3) particularly for the water body of high-density culture, the self-cleaning capacity of the water body is very limited due to large culture density and limited culture area, and diseases are caused by the deterioration of water quality at the middle and later stages of culture;
(4) the water quality stability is poor, and the quality and the safety of aquatic products are poor; the culture density and unit benefit are difficult to improve; the pollution problem of the aquaculture industry is increasingly prominent;
(5) the strain is low in load and quick and not lasting in release, and common bacterial liquid or powder needs to be frequently put in; the existing microbial preparation is difficult to be used in the flowing and open water environment.
Disclosure of Invention
In view of the above, the present invention aims to provide a preparation method of a microbial agent carrier material and an application thereof in high-density aquaculture, wherein the microbial agent carrier material decomposes pollutants in a water body under artificial promotion engineering conditions, reduces the concentrations of ammonia nitrogen and nitrite in the water body of an aquaculture pond, and repairs a polluted environment.
The technical scheme adopted by the invention for realizing the purpose is as follows:
a preparation method of a microbial agent carrier material comprises the following steps: bamboo charcoal pretreatment, culture medium preparation, bacterial liquid preparation, bacterial mud preparation, microorganism immobilization and sodium alginate wrapping;
s1, bamboo charcoal pretreatment: a1, putting bamboo charcoal into a drying oven at 105 ℃ for drying for 4 hours; a2, packaging the dried bamboo charcoal into 1L triangular bottles, wherein each bottle contains 150g of bamboo charcoal; a3, injecting 250ml of distilled water into each a2 triangular flask, and covering a plastic septum; a4, placing the triangular flask into a high-pressure steam sterilization pot, and sterilizing at 121 ℃ for 20 min; a5, taking out the triangular flask a4, and pouring water out of the triangular flask for later use after the triangular flask is cooled to room temperature;
s2, preparing a culture medium: b1, preparing an LB broth culture medium, sequentially filling the prepared LB broth culture medium into a 500ml triangular flask and a 250ml triangular flask, covering a plastic film, and marking an LB on the flask body;
b2, placing the LB broth culture medium which is subpackaged in the triangular flask into a high-pressure steam sterilization pot, and sterilizing at the temperature of 121 ℃ for 20 min;
s3, preparing a bacterial liquid: c1, remove the seed from the freezer, inoculate 1 loop to the sterilized 250ml Erlenmeyer flask medium in S2 with 10. mu.l of inoculating loop. Placing the inoculated culture medium in a shaking table at 32 ℃ and 200r/min for 24 h; c2, taking the culture medium after shake culture out of the shaker, and inoculating the culture medium into the sterilized 500ml triangular flask culture medium in S2 according to the inoculation amount of 10 percent; c3, placing the culture medium inoculated with the c2 at 32 ℃ for shake cultivation at 200r/min for 24 hours, and observing the growth condition of the bacteria and whether the bacteria are polluted;
s4, preparing bacterial sludge and fixing microorganisms: d1, taking out the sterilized centrifugal tubes, pouring the bacterial liquid amplified and cultured in S3 into the corresponding centrifugal tubes in sequence at 5000r/min, centrifuging for 5min, and removing supernatant to obtain bacterial sludge; d2, taking a triangular flask which is subjected to dry heat sterilization, re-suspending the bacterial sludge by using sterilized distilled water, adjusting OD600 of the re-suspended bacterial sludge to 1.2 +/-0.1, pouring bacterial liquid after adjusting OD600 into the triangular flask containing the bamboo charcoal sterilized in S1, pouring 250ml of the bacterial liquid into each bottle, covering a plastic film, putting the bottles into a shaking table at 32 ℃, and culturing for 24 hours at 200 r/min;
s5, wrapping with sodium alginate: e1, preparing a 4% sodium alginate solution, dissolving the solution in a water bath kettle at 80 ℃, taking out the solution after dissolving, and cooling the solution for later use; e2, preparing 1.38% calcium chloride aqueous solution; e3, taking out the triangular flask in the S4 after fixed culture from the shaking table, and pouring out the supernatant; e4, pouring the cooled 4% sodium alginate solution into a triangular flask filled with bamboo charcoal in S4, uniformly mixing and wrapping, taking out one particle of bamboo charcoal wrapped with sodium alginate by using forceps, putting into 1.38% calcium chloride solution, standing, wrapping and curing for 4 h; e5, pouring the calcium chloride solution, taking out the bamboo charcoal wrapped in e4, and storing in a self-sealing bag to obtain the microbial agent carrier material.
Preferably, in S1, the bamboo charcoal is a porous material carrier, is a micron-sized three-dimensional honeycomb structure biological carbon material, and has a density of 1.05g/cm3The solid material of (a), the microbial load in the bamboo charcoal>10 hundred million cfu/g.
Preferably, in S2, the bacterial liquid is a composite microbial liquid, the types of strains in the bacterial liquid include QB-1, QX-1, PSB and lactic acid bacteria, and the specific microbial strains include photosynthetic bacteria, pseudomonas, nitrobacteria, denitrifying bacteria and lactic acid bacteria.
Preferably, in S5, the volume ratio of the 4% sodium alginate solution to the 1.38% calcium chloride aqueous solution is 1: 10.
The invention also discloses application of the microbial agent carrier material in high-density aquaculture.
Another object of the present invention is to provide a stable and sustainable solution for water purification and ecological conservation in high-density aquaculture environment, which solves the problem that the existing microbial preparation is difficult to be used in flowing and open water environment.
The technical scheme adopted by the invention for realizing the purpose is as follows:
the microbial agent carrier material prepared by the method. The slow-release oxygen-increasing material is put into a high-density aquaculture farm, the slow-release oxygen-increasing material just sinks to a water bottom floating mud layer to play a role, the mud and water cannot be lost and float away, the mud and water are treated simultaneously, the microbial strains are solidified on a honeycomb-like micron porous bamboo charcoal material carrier and are stably attached to the water bottom, a unique artificial microenvironment is provided for the beneficial microbial strains, and the effect of once putting and long-term effectiveness is achieved; the microbial agent carrier can decompose pollutants in the water body of a high-density aquaculture farm, reduce the concentration of ammonia nitrogen and nitrite in the water body of an aquaculture pond, restore the polluted environment and also assist in improving the biological activity of microorganisms by utilizing the microorganisms under the condition of artificially promoting engineering.
Preferably, the slow-release oxygen increasing material comprises an oxygen releasing agent, attapulgite, stearic acid and PVP.
Preferably, the microbial agent carrier material and the slow-release oxygen increasing material both have the density of 1.05g/cm3The solid material of (1).
Preferably, the mass ratio of the microbial agent carrier material to the slow-release oxygen-increasing material is 2: 1.
Preferably, the microbial agent carrier material is applied to improving the water body environment in high-density aquaculture.
Preferably, the microbial agent carrier material is applied to improving the culture density of farmers and reducing the occurrence of fish diseases in high-density aquaculture.
Compared with the prior art, the invention has the following beneficial effects:
(1) the invention relates to a preparation method of a microbial agent carrier material and application thereof in high-density aquaculture, which mainly uses microbial agent products (QB-1, QX-1, PSB and lactobacillus); the solid state is used as an auxiliary material (microorganism bottom-sinking slow-release material and slow-release oxygen-increasing material), a stable and sustainable water quality purification product and an ecological preservation solution are provided for a high-density aquaculture environment, the use of pesticides and antibiotics is greatly reduced, the difficult problems that the water quality stability is poor, the quality and the safety of aquatic products are poor, and the aquaculture density and unit benefit are difficult to improve are mainly solved, the conversion of the development mode of the aquaculture industry is accelerated, and a new development mode with green, low carbon and continuous development is created;
(2) micro-meterThe density of the biological agent carrier material is 1.05g/cm3The sludge just sinks to the floating sludge layer at the bottom of the water to play a role, and the sludge and the water are not lost and floated, so that the sludge and the water are treated at the same time; meanwhile, the slow-release oxygen increasing material is matched for application, so that the oxygen is released slowly and durably, and the water body is oxygenated continuously;
(3) the lactobacillus is a gram-positive bacterium which can generate lactic acid, the lactobacillus can be used as bait material, the intestinal microbial community structure of the fish can be effectively adjusted to be stable, meanwhile, the mixed bacteria can be inhibited, the utilization rate of the bait can be improved by the generated metabolite, meanwhile, the lactobacillus preparation can be added into the water body, the water quality can be effectively improved, and the fish disease can be reduced. The microbial agent carrier material can effectively regulate the stable structure of the intestinal microbial community of the fish, inhibit the growth of mixed bacteria, improve the utilization rate of bait, reduce nitrite and the like;
(4) the microbial agent carrier material has the advantages of improving the water body environment, inhibiting the growth of pathogenic bacteria, recovering a clean water environment, degrading toxic and harmful substances, increasing the oxygen content, effectively avoiding the situation that the healthy growth of water body animals is hindered due to overhigh ammonia content, forming a specific population when beneficial microorganisms reach a certain amount in a water body, inhibiting the growth of harmful microorganisms by the beneficial microorganisms in the survival competition, improving the quality of the water body to a great extent, keeping the water body in a good stable state, and solving the problem of the discharge of traditional cultured pesticides, antibiotics and wastewater from the source;
(5) the invention takes the solid microorganism bottom-sinking material as the carrier and the liquid microbial inoculum as the water aqua, solves the problems of the traditional aquaculture pesticide, antibiotics, waste water discharge, environmental pollution destruction and the like from the source, realizes the ecological and naturalization of the aquaculture, really recovers the self-cleaning capability of the water body, and achieves the fundamental aims of long-term treatment, long-term cleanness and excellent ecology;
(6) according to the invention, microbial strains are solidified on the honeycomb-like micron porous bamboo charcoal material carrier, and the material is stably attached when being settled to the water bottom, so that the problem that the existing microbial preparation is difficult to use in flowing and open water body environments is solved, a unique artificial microenvironment is provided for beneficial microbial flora, and the effect of one-time release and long-term effectiveness is achieved;
(7) the invention utilizes harmless biological agents to inhibit pathogenic bacteria such as vibrio and the like in aquaculture, realizes large-scale culture in factories, increases the yield and reduces the feed coefficient; the microorganism in the natural environment or the specific microorganism is used for decomposing pollutants under the condition of artificially promoting engineering, reducing the concentration of ammonia nitrogen and nitrite in the water body of the culture pond and restoring the polluted environment.
Drawings
FIG. 1 shows the ammonia nitrogen detection result of the first experiment in the case of water quality purification of the culture water body of Acipenser sinensis grain, which should be collected in Hubei province.
FIG. 2 is a result of nitrite detection in the first experiment in case of water quality purification of Acipenser sinensis grain aquaculture water in Hubei province.
FIG. 3 is a second experimental ammonia nitrogen detection result in case of water quality purification of the culture water body of the Acipenser sinensis paddy in Hubei province.
FIG. 4 is a second experimental nitrite detection result in case of water quality purification of sturgeon grain aquaculture water in Hubei province.
FIG. 5 shows the ammonia nitrogen detection result of the fish tank experiment.
Detailed Description
The invention is further described with reference to the following drawings and detailed description. The examples are given solely for the purpose of illustrating the invention and are not intended to limit the scope of the invention. The examples provided below serve as a guide for further modifications by a person skilled in the art and do not constitute a limitation of the invention in any way.
Example 1:
a preparation method of a microbial agent carrier material comprises the following steps: bamboo charcoal pretreatment, culture medium preparation, bacterial liquid preparation, bacterial mud preparation, microorganism immobilization and sodium alginate wrapping;
s1, bamboo charcoal pretreatment: a1, weighing 750g of bamboo charcoal by using a balance, and drying the bamboo charcoal in an oven at 105 ℃ for 4 h; a2, subpackaging the dried bamboo charcoal into 1L triangular bottles, 150g each bottle, and subpackaging 4 bottles; a3, injecting 250ml of distilled water into each triangular flask, and covering a plastic diaphragm; a4, placing the triangular flask into a high-pressure steam sterilization pot, and sterilizing at 121 ℃ for 20 min; a5, taking out the triangular flask, and pouring water out of the triangular flask for later use after the triangular flask is cooled to room temperature;
s2, preparing a culture medium: b1, preparing 1.1L of LB broth culture medium, and sequentially filling 1.1L of LB broth culture medium into 4 triangular flasks with 500ml, wherein each flask is filled with 200 ml; 2 triangular flasks of 500ml are filled with 100ml of the mixture in each flask; 2 triangular bottles of 250ml are filled in 50ml of each bottle, a plastic film is covered on each bottle, and the bottle body is marked with LB; b2, placing the LB broth culture medium into an autoclave for sterilization at the temperature of 121 ℃ for 20 min;
s3, preparing a bacterial liquid: c1, remove the seed from the freezer, inoculate 1 loop with 10. mu.l of inoculating loop into sterilized medium (50 ml). Placing the inoculated culture medium in a shaking table at 32 ℃ and 200r/min for 24 h; c2, taking out the culture medium after shake culture from a shaking table, inoculating the culture medium into 100ml and 200ml sterilized triangular bottles according to the inoculation amount of 10%, and marking corresponding labels; c3, placing the inoculated culture medium in a shaking table at 32 ℃ and 200r/min for 24 hours, and observing the growth condition of the bacteria and whether the bacteria are polluted;
s4, preparing bacterial sludge and fixing microorganisms: d1, taking out the sterilized centrifugal tubes, pouring the bacterial liquid amplified and cultured in S3 into the corresponding centrifugal tubes in sequence, centrifuging for 5min at a speed of 5000r/min, removing supernatant to obtain bacterial sludge, and recording the volume of each thallus in sequence; d2, taking a triangular flask which is subjected to dry heat sterilization, re-suspending the bacterial sludge by using sterilized distilled water, adjusting OD600 of the re-suspended bacterial sludge to 1.2 +/-0.1, pouring bacterial liquid after adjusting OD600 into the triangular flask containing the bamboo charcoal sterilized in S1, pouring 250ml of the bacterial liquid into one bottle, covering a plastic film, putting the bottle into a shaking table at 32 ℃, and culturing for 24 hours at 200 r/min;
s5, wrapping with sodium alginate: e1, adding 200ml of water into 8g of sodium alginate, preparing 200ml of 4% sodium alginate, dissolving the solution in a water bath kettle at 80 ℃, taking out the solution after dissolution, and cooling the solution for later use; e2, preparing 2L of 1.38% calcium chloride aqueous solution; e3, taking out the triangular flask after fixed culture from the shaking table, and pouring out the supernatant; e4, pouring cooled sodium alginate into a triangular flask filled with bamboo charcoal, mixing uniformly, putting one granule of bamboo charcoal into calcium chloride solution with tweezers, and standing for wrapping for 4 h; e5, pouring the calcium chloride solution, taking out the wrapped bamboo charcoal, and storing in a self-sealing bag to obtain the microbial agent carrier material.
Example 2:
in the case of water quality purification of the sturgeon valley culture water in hubei province, the microbial inoculum carrier material prepared in the embodiment 1 is matched with the slow-release oxygen increasing material to be applied to sturgeon valley culture projects in Qingjiang (province) and penaeus vannamei culture projects in Qingjiang (Shenzhen), and the steps are as shown in fig. 1-4: the application effect is ideal, after the immobilized microorganism product is put into use, the reduction of ammonia nitrogen, nitrite and the like in water is obvious, and the oxygen content in the water body is obviously increased.
The development of intensive production mode of aquaculture industry goes through the stages of pond, open flowing water pond, net cage mode and the like, and now enters the development stage of industrial recirculating aquaculture. Compared with the first three modes, the industrial culture has the advantages that the dependence degree on the environment and resources is reduced; the influence degree on the environment is reduced, and the industrial culture is the development direction of the aquaculture industry. However, domestic recirculating aquaculture enterprises lack relevant experience, professional recirculating aquaculture technicians and referable industrial recirculating aquaculture technical data, the aquaculture density is not easy to control, and the water quality cannot be effectively regulated and controlled. The strains and preparations developed by companies can well control the concentration of ammonia nitrogen, nitrite and the like in water which are toxic to fish, improve the breeding density of farmers and increase the economic benefit.
In the embodiment, QB-1 microorganism bottom-sinking slow-release materials are added into a culture pond according to the dosage of 0.1 kg/cubic water, and the indexes of water temperature, pH, dissolved oxygen, ammonia nitrogen, nitrate and nitrite in the water are periodically detected to monitor the water quality. In this example a total of two monitoring runs were performed.
The results of the two tests show that the ammonia nitrogen and nitrite in the experimental group added with the microbial bottom-settling material are obviously lower than those in the blank group. And the difference between the nitrite groups of the two experimental groups is obviously smaller than that of the blank group. The invention has obvious effect in aquaculture application, can reduce the pollution of aquaculture water and improve the aquaculture density.
Example 3:
the ammonia nitrogen detection in the fish tank experiment is carried out, as shown in fig. 5, 3 50L fish tanks are selected in the experiment and are randomly divided into a 50g QB-1 group, a 100gQB-1 group and a blank group, 50g of microorganism bottom-sinking slow-release material QB-1 is added in the 50g QB-1 group, 100gQB-1 group of microorganism bottom-sinking slow-release material QB-1 is added in the blank group, and 50g of common biochar is added in the blank group to serve as blank control. The experimental result shows that compared with the blank fish tank added with the microorganism bottom-sinking slow-release material QB-1, the ammonia nitrogen in the water can maintain a lower level, the water quality of the fish tank is ensured, and the threshold of the breeding technology is reduced. Because the ammonia nitrogen can be controlled at a lower level, the water changing frequency of the fish tank can be reduced, and the culture workload is reduced.
The microbial agents of the present invention are currently applied in a number of aquaculture projects, for example: the application effect is ideal, after the microbial inoculum product is put into use, the reduction of ammonia nitrogen, phosphorus, nitrite and the like in water is obvious, and the oxygen content in the water body is obviously increased.
The long-term reasonable application of the microecological preparation can ensure that the ecological advantages of beneficial microbial floras are formed in the culture water area, and the beneficial cycle effect of promoting the healthy development of culture activities is achieved. The microorganisms living in the pond or the specific microorganisms are used for decomposing pollutants under the condition of artificially promoting engineering, reducing the concentration of ammonia nitrogen and nitrite in the water body of the culture pond and repairing the polluted environment.
The application of bioremediation technology is to ensure sustainable utilization and industrial sustainable development of aquaculture resources, and is an important field of biotechnology research and development at present, wherein development and application of environmental bioremediation technology are key points of environmental and ecological technology research and development. Bioremediation technology has been widely used abroad to treat water area environment as an efficient and energy-saving environmental protection technology. With the increasing prominence of the pollution problem of the aquaculture industry, people explore methods and ways for improving the culture environment and purifying the culture water quality to the utmost extent.
The working principle of the invention is as follows: mainly uses microbial agent products (QB-1, QX-1, PSB and lactobacillus); the solid state is used as an auxiliary material (microorganism bottom-sinking slow-release material and slow-release oxygen-increasing material), a stable and sustainable water quality purification product and an ecological preservation solution are provided for a high-density aquaculture environment, the use of pesticides and antibiotics is greatly reduced, the difficult problems that the water quality stability is poor, the quality and the safety of aquatic products are poor, and the aquaculture density and unit benefit are difficult to improve are mainly solved, the conversion of the development mode of the aquaculture industry is accelerated, and a new development mode with green, low carbon and continuous development is created; because the density of the microbial agent carrier material is 1.05g/cm3The sludge just sinks to the floating sludge layer at the bottom of the water to play a role, and the sludge and the water are not lost and floated, so that the sludge and the water are treated at the same time; meanwhile, the slow-release oxygen increasing material is matched for application, so that the oxygen is released slowly and durably, and the water body is oxygenated continuously; the microbial agent carrier material can effectively regulate the stable structure of the intestinal microbial community of the fish, inhibit the growth of mixed bacteria, improve the utilization rate of bait, reduce nitrite and the like; the microbial agent carrier material has the advantages of improving the water body environment, inhibiting the growth of pathogenic bacteria, recovering a clean water environment, degrading toxic and harmful substances, increasing the oxygen content, effectively avoiding the situation that the healthy growth of water body animals is hindered due to overhigh ammonia content, forming a specific population when beneficial microorganisms reach a certain amount in a water body, inhibiting the growth of harmful microorganisms by the beneficial microorganisms in the survival competition, improving the quality of the water body to a great extent, keeping the water body in a good stable state, and solving the problem of the discharge of traditional cultured pesticides, antibiotics and wastewater from the source; solid microorganism sediment materials are used as carriers, liquid microbial agents are used as water agents, the problems of traditional aquaculture pesticides, antibiotics, waste water discharge, environmental pollution destruction and the like are solved from the source, the ecological and naturalization of aquaculture is realized, the self-cleaning capacity of water is really recovered, and the fundamental aims of long-term treatment, long-term cleanness and excellent ecology are achieved; by solidifying on the honeycomb-like micron porous bamboo charcoal material carrier, the material is stably attached when being settled to the water bottom, thereby solving the problem that the existing microbial preparation is difficult to be used in flowing and open water body environment, providing a unique artificial microenvironment for beneficial microbial flora and achieving the aim ofThe effect is long and effective after one-time putting; the harmless biological agent is used for inhibiting pathogenic bacteria such as vibrio and the like in aquaculture, so that the large-scale culture of a factory is realized, the yield is increased, and the bait coefficient is reduced; the microorganism in the natural environment or the specific microorganism is used for decomposing pollutants under the condition of artificially promoting engineering, reducing the concentration of ammonia nitrogen and nitrite in the water body of the culture pond and restoring the polluted environment.
The above-mentioned embodiments are only preferred embodiments of the present invention, and the scope of the claims of the present invention should not be limited by the above-mentioned embodiments, because the modifications, equivalent variations, improvements, etc. made in the claims of the present invention still fall within the scope of the present invention.
Claims (11)
1. A preparation method of a microbial agent carrier material comprises the following steps: bamboo charcoal pretreatment, culture medium preparation, bacterial liquid preparation, bacterial mud preparation, microorganism immobilization and sodium alginate wrapping; s1, bamboo charcoal pretreatment: a1, putting bamboo charcoal into a drying oven at 105 ℃ for drying for 4 hours; a2, packaging the dried bamboo charcoal into 1L triangular bottles, wherein each bottle contains 150g of bamboo charcoal; a3, injecting 250ml of distilled water into each a2 triangular flask, and covering a plastic septum; a4, placing the triangular flask into a high-pressure steam sterilization pot, and sterilizing at 121 ℃ for 20 min; a5, taking out the triangular flask a4, and pouring water out of the triangular flask for later use after the triangular flask is cooled to room temperature; s2, preparing a culture medium: b1, preparing an LB broth culture medium, sequentially filling the prepared LB broth culture medium into a 500ml triangular flask and a 250ml triangular flask, covering a plastic film, and marking an LB on the flask body; b2, placing the LB broth culture medium which is subpackaged in the triangular flask into a high-pressure steam sterilization pot, and sterilizing at the temperature of 121 ℃ for 20 min; s3, preparing a bacterial liquid: c1, remove the seed from the freezer, inoculate 1 loop to the sterilized 250ml Erlenmeyer flask medium in S2 with 10. mu.l of inoculating loop. Placing the inoculated culture medium in a shaking table at 32 ℃ and 200r/min for 24 h; c2, taking the culture medium after shake culture out of the shaker, and inoculating the culture medium into the sterilized 500ml triangular flask culture medium in S2 according to the inoculation amount of 10 percent; c3, placing the culture medium inoculated with the c2 at 32 ℃ for shake cultivation at 200r/min for 24 hours, and observing the growth condition of the bacteria and whether the bacteria are polluted; s4, preparing bacterial sludge and fixing microorganisms: d1, taking out the sterilized centrifugal tubes, pouring the bacterial liquid amplified and cultured in S3 into the corresponding centrifugal tubes in sequence at 5000r/min, centrifuging for 5min, and removing supernatant to obtain bacterial sludge; d2, taking a triangular flask which is subjected to dry heat sterilization, re-suspending the bacterial sludge by using sterilized distilled water, adjusting OD600 of the re-suspended bacterial sludge to 1.2 +/-0.1, pouring bacterial liquid after adjusting OD600 into the triangular flask containing the bamboo charcoal sterilized in S1, pouring 250ml of the bacterial liquid into each bottle, covering a plastic film, putting the bottles into a shaking table at 32 ℃, and culturing for 24 hours at 200 r/min; s5, wrapping with sodium alginate: e1, preparing a 4% sodium alginate solution, dissolving the solution in a water bath kettle at 80 ℃, taking out the solution after dissolving, and cooling the solution for later use; e2, preparing 1.38% calcium chloride aqueous solution; e3, taking out the triangular flask in the S4 after fixed culture from the shaking table, and pouring out the supernatant; e4, pouring the cooled 4% sodium alginate solution into a triangular flask filled with bamboo charcoal in S4, uniformly mixing and wrapping, taking out one particle of bamboo charcoal wrapped with sodium alginate by using forceps, putting into 1.38% calcium chloride solution, standing, wrapping and curing for 4 h; e5, pouring the calcium chloride solution, taking out the bamboo charcoal wrapped in e4, and storing in a self-sealing bag to obtain the microbial agent carrier material.
2. The microbial inoculant carrier material of claim 1, wherein: s1, the bamboo charcoal is a porous material carrier, is a micron-sized three-dimensional honeycomb structure biological carbon material and has a density of 1.05g/cm3The solid material of (a), the microbial load in the bamboo charcoal>10 hundred million cfu/g.
3. The microbial inoculant carrier material of claim 1, wherein: in S2, the bacterial liquid is a compound microorganism bacterial liquid, the types of the strains in the bacterial liquid comprise QB-1, QX-1, PSB and lactic acid bacteria, and the specific microorganism bacterial comprises photosynthetic bacteria, pseudomonas, nitrobacteria, denitrifying bacteria and lactic acid bacteria.
4. The microbial inoculant carrier material of claim 1, wherein: in S5, the volume ratio of a 4% sodium alginate solution to a 1.38% calcium chloride aqueous solution is 1: 10.
5. The use of the microbial inoculant carrier material of claim 1 in high density aquaculture.
6. Use according to claim 5, characterized in that: the microbial agent carrier material is used together with a slow-release oxygen increasing material.
7. The microbial inoculant carrier material and the slow-release oxygen increasing material as claimed in claim 6, wherein: the microbial agent carrier material and the slow-release oxygen increasing material both have the density of 1.05g/cm3The solid material of (1).
8. The slow release oxygen increasing material according to claim 6, wherein: the slow-release oxygen increasing material comprises an oxygen releasing agent, attapulgite, stearic acid and PVP.
9. The microbial inoculant carrier material and the slow-release oxygen increasing material as claimed in claim 6, wherein: the mass ratio of the microbial agent carrier material to the slow-release oxygen increasing material is 2: 1.
10. The use of the microbial inoculant carrier material of claim 1 for modifying the environment of a body of water in high density aquaculture.
11. The use of the microbial inoculant carrier material of claim 1 for increasing farmer density and reducing fish disease occurrence in high density aquaculture.
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