CN114836333A - Blue algae purifying agent and preparation and application thereof - Google Patents

Blue algae purifying agent and preparation and application thereof Download PDF

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CN114836333A
CN114836333A CN202110140059.0A CN202110140059A CN114836333A CN 114836333 A CN114836333 A CN 114836333A CN 202110140059 A CN202110140059 A CN 202110140059A CN 114836333 A CN114836333 A CN 114836333A
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谷序文
曹芬
樊俊
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Wuhan Yuqingjia Ecological Technology Co.,Ltd.
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Abstract

The invention discloses a blue algae purifying agent and a preparation method thereof, the invention firstly prepares seed liquid of bacillus cereus and bacillus subtilis respectively, inoculates the two seed liquids into a fermentation tank simultaneously for liquid mixed fermentation, the fermentation culture adopts a segmented temperature control method, fermentation is carried out at different temperatures in the early stage, the middle stage and the later stage of the fermentation, coconut oil is added into the obtained fermentation liquor after the fermentation is finished, the mixture is mixed and stirred uniformly to obtain fermentation mixed liquor, the mixed liquor is concentrated, thalli are dried, detected and packaged, and the blue algae purifying agent is obtained. The invention also discloses a blue algae purifying agent obtained by using the method and application of the blue algae purifying agent in improving the water quality of aquaculture water. The invention adopts a segmented temperature control method in fermentation, effectively improves the number of viable bacteria, increases the spore rate by adding coconut oil after the liquid fermentation is finished, enhances the stress resistance of strains, ensures the stability of product quality, and has good effect when the obtained blue algae purifying agent is used for improving the water quality of the culture water body.

Description

Blue algae purifying agent and preparation and application thereof
Technical Field
The invention relates to a preparation method of a blue algae purifying agent, and also relates to the blue algae purifying agent prepared by the method, and further relates to the application of the blue algae purifying agent in ecological improvement of blue algae bloom and treatment of modern agriculture and water environment.
Background
Water is the environment on which the aquaculture animals depend to live, and the quality of the water directly influences the growth and development of the aquaculture animals and the economic benefit of the aquaculture.
The blue algae is a common harmful algae in culture water, mainly comprises microcystis, spirulina, anabaena, Oscillatoria and the like, wherein the microcystis is the most typical and most harmful.
Blue-green algae is easy to propagate in a large amount in eutrophic water, a sticky and fishy blue-green film is formed on the surface of the water, the blue-green algae can be fully distributed in the whole aquaculture water when the blue-green algae is serious, the blue-green algae has high growth speed and poor stability, and easily dies due to aging, the dead algae are decomposed by bacteria, and a large amount of dissolved oxygen in the water is consumed, so that the fish is anoxic. In addition, most of the blue algae have algae toxins, wherein the most studied algae toxins can directly cause poisoning and death of cultured animals, and the algae toxins have carcinogenicity and pose great threat to human health in terms of liver toxicity and neurotoxicity. The cyanobacterial bloom not only destroys the culture ecological environment and restricts the economic benefit of aquaculture, but also seriously harms the health of human beings.
At present, the method for treating the cyanobacterial bloom by the culture water body mainly comprises a physical method, a chemical method and a biological method. The physical methods comprise manual fishing, flocculation, hydrodynamic control and the like, the safety is good, but the engineering quantity is large, the operation is not easy, the blue algae is easy to erupt again, and the treatment of symptoms and root causes is not fundamental. The chemical method mainly kills the blue algae cells by putting chemical algicide (copper sulfate, potassium permanganate and the like), has no specificity and specificity, can damage fishes or other aquatic organisms while killing the blue algae, destroys the culture ecology, and easily causes secondary pollution of water and is not paid for by putting chemical medicine. The method for biologically treating the blue algae mainly comprises the steps that the aquatic plants control the blue algae, the filter-feeding aristichthys eats granular blue algae, beneficial algae inhibit the blue algae and the like, and is a relatively economic and safe algae control mode.
The growth and the propagation of the blue algae are controlled by cultivating aquatic plants to compete with the blue algae for nutrient substances, putting excessive bighead carp to prey on granular blue algae and the like, and the modes of regulating and controlling the harmful blue algae by utilizing large-scale aquatic animals and plants have the advantages of ecology and environmental protection, but have slow effect and high cost. In comparison, the microbial algae control technology has the advantages of strong specificity, low cost, easiness in operation, small influence on the environment, safety, high efficiency and the like, and is the most promising mode for regulating and controlling the blue-green algae.
At the present stage, the research of controlling the blue-green algae in the aquaculture water body by using the microecological preparation is not many, the research is mainly focused on the theoretical research of a microorganism algae dissolving mechanism and the like, the actual application reports are few, and the microecological preparation product for purifying the blue-green algae in the water body is almost not available. Mainly because the indigenous microorganisms in the natural water have strong competition, the algae-lysing bacteria are difficult to become dominant species and maintain a certain amount, and the growth and the algae-lysing activity of the algae-lysing bacteria are easily influenced by physicochemical conditions and biological factors in the water environment, so the actual algae-lysing effect is poor. In addition, most algae-lysing bacteria are strains separated and cultured by organic culture media such as meat extract, and the like, so that the strain culture cost is greatly consumed in the actual application, and the nutrient load of a water body is additionally increased.
The invention is mainly used for developing a high-efficiency microecological preparation capable of purifying the blue algae in the water body.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provide a preparation method of a microecological preparation for purifying blue algae, which has the advantages of low production cost, high viable count and spore rate of products and strong stress resistance. The invention also discloses a cyanobacteria purifying agent prepared by the method and the application of the purifying agent in the improvement of cyanobacteria water.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a preparation method of a blue algae purifying agent comprises the following steps:
(a) preparing seed liquid of Bacillus cereus and Bacillus subtilis;
(b) inoculating the seed liquid of the two strains into the same liquid fermentation culture medium for mixed fermentation culture, adopting a segmented temperature control method in the fermentation culture process, fermenting at different temperatures in the early stage, the middle stage and the later stage of fermentation, wherein the fermentation temperature is higher in the early stage of fermentation, the fermentation temperature is lower in the middle stage of fermentation, and the fermentation temperature is increased in the later stage of fermentation;
in the early stage of fermentation, the thalli are in a lag phase and in an early logarithmic growth phase, and the higher tank temperature is favorable for accelerating germination of the thalli and shortening the lag phase time; in the middle stage of fermentation, when thalli enter a period of logarithmic growth exuberance, the thalli generate heat seriously due to metabolism, local overhigh temperature is easy to appear, and the control temperature is properly reduced to avoid the thalli from aging and dying due to overheating; in the later stage of fermentation, the temperature of the tank is increased, so that the fermentation time is prolonged, and the number of produced bacteria is increased.
(c) After fermentation, adding coconut oil into the fermentation liquor obtained in the step (b), mixing and stirring uniformly to obtain a fermentation mixed liquor, concentrating the mixed liquor, drying the thalli, detecting and packaging to obtain the blue algae purifying agent.
Wherein the inoculation amounts of the Bacillus cereus and Bacillus subtilis liquid in the step (b) are equal and are 3-8% of the weight of the liquid fermentation medium; the fermentation culture conditions are as follows: the initial pH value is 7.0-7.5, the rotating speed is 150-180 r/min, the tank pressure is 0.04-0.06MPa, the fermentation time is 24-28 h, the temperature is controlled at 35-39 ℃ for 1-6 h, the temperature is controlled at 28-32 ℃ for 6-20 h, and the temperature is controlled at 33-37 ℃ from 20h to the end of fermentation; the liquid fermentation medium consists of the following components: 4-6% of cyanobacteria powder, 0.2-0.5% of ammonium sulfate, 0.1-0.2% of dipotassium hydrogen phosphate, 0.05% of magnesium sulfate, 0.05% of ferrous sulfate, 0.01% of manganese sulfate, 0.01% of sodium chloride and the balance of water; after the liquid fermentation medium is prepared according to the formula, the pH value is adjusted to 7.0-7.5.
Wherein, the inoculation amount of the Bacillus cereus and the Bacillus subtilis in the step (b) is preferably 5 percent of the weight of the liquid fermentation medium; the fermentation culture conditions are preferably: the initial pH value is 7.5, the rotating speed is 180r/min, the tank pressure is 0.04-0.06MPa, the fermentation time is 26h, the temperature is controlled at 37 ℃ for 1-6 h, the temperature is controlled at 30 ℃ for 6-20 h, and the temperature is controlled at 35 ℃ from 20h to the end of fermentation; the content of each component of the liquid fermentation medium is preferably as follows: according to the weight percentage, the blue algae powder is 5 percent, the ammonium sulfate is 0.3 percent, the dipotassium phosphate is 0.2 percent, the magnesium sulfate is 0.05 percent, the ferrous sulfate is 0.05 percent, the manganese sulfate is 0.01 percent, the sodium chloride is 0.01 percent, and the balance is water; after the liquid fermentation medium is prepared according to the formula, the pH value is adjusted to 7.5.
Wherein, the volume of the coconut oil added into the fermentation liquor after the fermentation in the step (c) is finished is 8-12% of the volume of the fermentation liquor, and the mixing and stirring time is 1-2 hours. Preferably, the volume of coconut oil added is 10% of the volume of the fermentation broth, and the mixing and stirring time is 2 hours. The method for concentrating the mixed solution is to collect thalli after plate-frame filtration; the method for drying the thallus is a conventional physical drying method.
The cyanobacteria purifying agent obtained by the method and the application of the cyanobacteria purifying agent in the improvement of the aquatic environment of the culture water body also belong to the protection scope of the invention.
Compared with the prior art, the invention has the following advantages:
(1) the invention adopts the composite microecological preparation to decompose blue algae and degrade algal toxins, and has the advantages of high efficiency, environmental protection and safety to cultured animals compared with the traditional chemical algae killing product;
(2) the preparation of the blue algae purifying agent adopts a high-density liquid mixed bacteria fermentation process. The fermentation liquor of the technology takes the cyanobacteria powder as a substrate, thereby reducing the production cost and simultaneously improving the quick adaptability and effectiveness of the preparation to the environment when in use. The viable count is improved by combining mixed fermentation with a segmented temperature control technology, 10% coconut oil in volume is added after fermentation is finished, and the coconut oil is mixed and stirred for 2 hours, so that the spore rate is improved, the stress resistance of a strain is enhanced, and the stability of the product quality is ensured;
(3) the cyanobacteria purifying agent can effectively remove cyanobacteria and degrade cyanobacteria toxin, and has the advantages of simple operation, low cost and easy popularization and application.
The details of the present invention can be obtained from the following description and the attached drawings.
Drawings
FIG. 1 is a process route diagram of the liquid mixed fermentation method of the cyanobacteria purifying agent.
FIG. 2 is a graph showing the comparison of viable count growth in fermentation solutions under different temperature control modes.
Detailed Description
The advantages and features of the present invention will become more apparent from the following description of the embodiments of the invention, taken in conjunction with the accompanying drawings and examples. These examples are illustrative only and do not limit the scope of the present invention in any way. It will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention, and that such changes and modifications may be made without departing from the spirit and scope of the invention.
Experimental Material
The strains used in the invention are purchased from agricultural microorganism center of China Committee for culture Collection of microorganisms, and the product number of Bacillus cereus is as follows: ACCC03320, the product number of the Bacillus subtilis is as follows: ACCC 03538.
EXAMPLE 1 preparation of cyanobacteria scavenger
1. Respectively inoculating commercially available Bacillus cereus and Bacillus subtilis strains in triangular flasks filled with LB liquid culture medium for culture at 37 deg.C for 24 hr to obtain seed solutions of the two strains.
2. And (3) inoculating the two seed bacterial liquids obtained in the step (1) to the same liquid fermentation medium according to 5% of the weight of the liquid fermentation medium for mixed bacterial fermentation culture. The liquid fermentation medium consists of the following components: according to the weight percentage, the blue algae powder is 5 percent, the ammonium sulfate is 0.3 percent, the dipotassium phosphate is 0.2 percent, the magnesium sulfate is 0.05 percent, the ferrous sulfate is 0.05 percent, the manganese sulfate is 0.01 percent, the sodium chloride is 0.01 percent, and the balance is water. After the liquid fermentation medium is prepared according to the formula, the pH value is adjusted to 7.5. In the fermentation culture process, a segmented temperature control method is adopted, the initial pH value is 7.5, the rotating speed is 180r/min, the tank pressure is 0.04-0.06MPa, the fermentation time is 26h, the temperature is controlled at 37 ℃ for 1-6 h, the temperature is controlled at 30 ℃ for 6-20 h, and the temperature is controlled at 35 ℃ after 20h and fermentation.
3. And (3) after the fermentation is finished, adding coconut oil into the fermentation liquor obtained in the step (2), wherein the volume of the added coconut oil is 10% of the volume of the fermentation liquor, and uniformly mixing and stirring for 2 hours to obtain a fermentation mixed liquor.
4. Concentrating the mixed solution, drying the thalli, detecting and packaging to obtain the blue algae purifying agent.
Example 2 preparation of a cyanobacteria scavenger
1. Respectively inoculating commercially available Bacillus cereus and Bacillus subtilis strains in triangular flasks filled with LB liquid culture medium for culture at 37 deg.C for 24 hr to obtain seed solutions of the two strains.
2. And (3) inoculating the two kinds of seed bacterial liquids obtained in the step (1) to the same liquid fermentation culture medium according to 8% of the weight of the liquid fermentation culture medium for mixed bacterial fermentation culture. The liquid fermentation medium consists of the following components: the blue algae powder comprises, by weight, 6% of blue algae powder, 0.2% of ammonium sulfate, 0.2% of dipotassium phosphate, 0.05% of magnesium sulfate, 0.05% of ferrous sulfate, 0.01% of manganese sulfate, 0.01% of sodium chloride and the balance of water. After the liquid fermentation medium is prepared according to the formula, the pH value is adjusted to 7.5. In the fermentation culture process, a segmented temperature control method is adopted, the initial pH value is 7.5, the rotating speed is 165r/min, the tank pressure is 0.04-0.06MPa, the fermentation time is 24h, the temperature is controlled for 1-6 h at 35 ℃, 6-20 h at 32 ℃, and the temperature is controlled for 20h until the end of fermentation at 33 ℃.
3. And (3) after the fermentation is finished, adding coconut oil into the fermentation liquor obtained in the step (2), wherein the volume of the added coconut oil is 8% of the volume of the fermentation liquor, and uniformly mixing and stirring for 2 hours to obtain a fermentation mixed liquor.
4. Concentrating the mixed solution, drying the thalli, detecting and packaging to obtain the blue algae purifying agent.
EXAMPLE 3 preparation of cyanobacteria scavenger
1. Respectively inoculating commercially available Bacillus cereus and Bacillus subtilis strains in triangular flasks filled with LB liquid culture medium for culture at 37 deg.C for 24 hr to obtain seed solutions of the two strains.
2. And (3) inoculating the two seed bacterial liquids obtained in the step (1) to the same liquid fermentation medium according to the weight of 3% of the liquid fermentation medium for mixed bacterial fermentation culture. The liquid fermentation medium consists of the following components: the blue algae powder comprises, by weight, 4% of blue algae powder, 0.5% of ammonium sulfate, 0.1% of dipotassium phosphate, 0.05% of magnesium sulfate, 0.05% of ferrous sulfate, 0.01% of manganese sulfate, 0.01% of sodium chloride and the balance of water. After the liquid fermentation medium is prepared according to the formula, the pH value is adjusted to 7.0. In the fermentation culture process, a segmented temperature control method is adopted, the initial pH value is 7.0, the rotating speed is 150r/min, the tank pressure is 0.04-0.06MPa, the fermentation time is 28h, the temperature is controlled at 39 ℃ for 1-6 h, the temperature is controlled at 28 ℃ for 6-20 h, and the temperature is controlled at 37 ℃ after 20h and fermentation are finished.
3. And (3) after the fermentation is finished, adding coconut oil into the fermentation liquor obtained in the step (2), wherein the volume of the added coconut oil is 12% of the volume of the fermentation liquor, and uniformly mixing and stirring for 1 hour to obtain a fermentation mixed liquor.
4. Concentrating the mixed solution, drying the thalli, detecting and packaging to obtain the blue algae purifying agent.
Preparation of blue algae scavenger without adding coconut oil in comparison example
1. Respectively inoculating commercially available Bacillus cereus and Bacillus subtilis strains in triangular flasks filled with LB liquid culture medium for culture at 37 deg.C for 24 hr to obtain seed solutions of the two strains.
2. And (3) inoculating the two seed bacterial liquids obtained in the step (1) to the same liquid fermentation medium according to 5% of the weight of the liquid fermentation medium for mixed bacterial fermentation culture. The liquid fermentation medium consists of the following components: according to the weight percentage, the blue algae powder is 5 percent, the ammonium sulfate is 0.3 percent, the dipotassium phosphate is 0.2 percent, the magnesium sulfate is 0.05 percent, the ferrous sulfate is 0.05 percent, the manganese sulfate is 0.01 percent, the sodium chloride is 0.01 percent, and the balance is water. After the liquid fermentation medium is prepared according to the formula, the pH value is adjusted to 7.5. In the fermentation culture process, a segmented temperature control method is adopted, the initial pH value is 7.5, the rotating speed is 180r/min, the tank pressure is 0.04-0.06MPa, the fermentation time is 26h, the temperature is controlled at 37 ℃ for 1-6 h, the temperature is controlled at 30 ℃ for 6-20 h, and the temperature is controlled at 35 ℃ after 20h and fermentation.
3. And after fermentation, concentrating the fermentation liquor, drying the thalli, detecting and packaging to obtain the blue algae purifying agent without adding coconut oil.
Experimental example 1 comparison of fermentation effects of a segmented temperature control method and a conventional temperature control method
1. Test method
Seed liquids of Bacillus cereus and Bacillus subtilis are prepared according to the step 1 in the embodiment 1, and in the step 2, three groups of tests are set for comparing fermentation effects, wherein:
and (3) treatment A: controlling the temperature of 30 ℃ in the whole fermentation process;
and (B) treatment: controlling the temperature of 37 ℃ in the whole fermentation process;
and C, treatment: and (3) adopting a segmented temperature control method to control the temperature in a segmented manner, wherein the temperature is controlled to be 37 ℃ within 1-6 h, 30 ℃ within 6-20 h and 35 ℃ within 20h after the fermentation is finished.
Extracting each group of fermentation liquor every 4h in the fermentation process to detect the number of viable bacteria, and obtaining a result shown in figure 2; the number of viable bacteria in each fermentation broth was measured at the end of fermentation and the results are shown in Table 1.
2. Test results
2.1 comparison of viable count growth during fermentation
As can be seen from FIG. 2, the temperature of the whole process of treatment A is controlled at a lower temperature of 30 ℃, the bacterial count in the early stage rises slowly, the bacterial count increases more quickly but highly limitedly and enters a stable stage quickly along with the log increase period, and the bacterial count can only be maintained at a certain level in the later stage of fermentation without further increase; the whole process of the treatment B is controlled at a higher temperature of 37 ℃, the bacterial count in the early stage rises quickly, but the bacterial strain ages quickly, and the growth rate of the bacterial count in the middle and later stages of fermentation is lower than that of the treatment A and the treatment C; the treatment C adopts a segmented temperature control method to control the temperature in a segmented mode, has obvious advantages compared with the treatment A and the treatment B, the bacteria number in the early stage rises quickly, meanwhile, the logarithmic growth with high speed can be realized in the middle stage, and the bacteria number in the later stage of fermentation is also slowly increased.
2.2 comparison of viable count in fermentation solutions under different temperature control modes
TABLE 1 comparison of viable count in fermentation solutions under different temperature control modes
Group A (30 ℃ C.) Group B (37 ℃ C.) Group C (short temperature control)
Viable count (Yi/mL) 79 70 114
The data in Table 1 show that the number of bacteria in the fermentation liquor in the segmented temperature control mode reaches 114 hundred million/mL, which is obviously superior to the number of bacteria in the fermentation liquor obtained by the conventional temperature control mode with the whole fixed temperature of 30 ℃ and 37 ℃.
Test example 2 comparison of spore formation rates for different post-treatments
1. Test method
The spore formation rates of the fermentation mixtures before concentration and drying were compared among inventive examples 1, 2, 3 and the control, and the results are shown in Table 2.
The ratio of spores (%) - (number of spores/total number of bacteria) × 100%
The detection culture medium is beef extract peptone culture medium
2. Test results
Table 2 shows the results of comparing the spore formation rates of the fermentation mixtures before concentration and drying in examples 1, 2 and 3 of the present invention and the comparative examples.
TABLE 2 comparison of sporulation rates for different post-treatments
Example 1 Example 2 Example 3 Comparative example
Number of viable bacteria (hundred million/ml) 114 108 105 114
Ratio of spores (%) 99.8 98.4 98.5 81.3
As shown in Table 2, the spore rates of the examples 1, 2 and 3 are higher than that of the comparative example by more than 20%, which shows that after the fermentation is completed, coconut oil is added into the fermentation liquid, and the mixture is uniformly mixed, so that the spore rate of the fermentation liquid can be obviously improved, the stress resistance of the strain is enhanced, and the stability of the product quality is ensured.
Test example 3 application of the present invention to practical aquaculture ponds
The test is carried out in the scientific farm of the farm for protecting the good country and the good fortune in the river and the summer of Wuhan city in 2020, 8 months, 20 days to 2020, 9 months and 9 days. In 8 and 9 months in summer, the temperature is high, the culture enters the middle and later stages, the pond accumulates more residual feed and excrement, the water eutrophication is serious, and the pond is a high season of blue algae. 4 grass carp aquaculture ponds A, B, C and D with serious blue-green algae condition are selected in the test, the water surfaces of the 4 ponds are all provided with a large number of blue-green algae floating, the pond water is aged and blackened, and the number of blue-green algae cells in the 4 ponds is detected to be more than 9 multiplied by 10 5 cell/mL, the content of microcystin (abbreviated as 'MC') exceeds 0.2 mu g/L.
The pond A is treated by applying the blue algae purifying agent in the example 1, and water is splashed in the whole pond after water is collected on the day of starting the test, wherein the using amount is 200 g/mu.m; the pond B is treated by applying a control example (a blue algae purifying agent without coconut oil is added), and water is splashed in the whole pond after water is completely collected on the day of starting the test, wherein the using amount is 200 g/mu.m; locally sprinkling copper sulfate to the pond C to kill algae in a conventional blue algae treatment mode, and changing 1/3 pond water after 2 days; pond D was an untreated control. During the test period, other input articles are not applied to each pool, only normal feeding and management are carried out, sampling is carried out for 1 time every 5 days, the change conditions of the number of the cyanobacteria cells and the MC content in the water body are detected, and the results are shown in a table 3:
TABLE 3 variation of the number of cyanobacterial cells and MC content
Figure BSA0000232072440000081
As can be seen from table 3, the indices for each pond varied during the test as follows:
after the blue algae purifying agent is applied to the pond A, the number of the blue algae is gradually reduced from 92006cell/mL to 1057cell/mL, the MC content is gradually reduced from 0.229 mu g/L to 0.002 mu g/L, the removal rates respectively reach 98.85% and 99.12%, and the blue algae can be continuously controlled to propagate and prevent the blue algae from erupting again.
After the pond B is fed with the blue algae purifying agent without coconut oil, the number of the blue algae is gradually reduced from 91893 cells/mL to 23547 cells/mL, the MC content is gradually reduced from 0.217 mu g/L to 0.058 mu g/L, the removal rates are 74.38% and 73.27%, the effect of removing the blue algae and the MC is obvious, but the effect is slightly worse than that of the blue algae purifying agent in the patent.
After the copper sulfate is applied to the pond C, the number of blue-green algae is rapidly reduced to a very low level, after 10 days, the blue-green algae rapidly grows again to form advantages, the number of the blue-green algae and the MC content are not reduced or increased reversely compared with those before treatment, and the culture environment is more deteriorated.
The pond D is not treated at all, and the number of blue algae and the MC content are continuously increased.
The preferred embodiments of the present invention have been described in detail, however, the present invention is not limited to the details of the above embodiments, and various simple modifications can be made to the technical solution of the present invention within the technical idea of the present invention, and these simple modifications are within the protective scope of the present invention.

Claims (10)

1. A preparation method of a blue algae purifying agent comprises the following steps:
(a) preparing seed liquid of Bacillus cereus and Bacillus subtilis;
(b) inoculating the seed liquid of the two strains into the same liquid fermentation culture medium for fermentation culture, adopting a segmented temperature control method in the fermentation culture process, fermenting at different temperatures in the early stage, the middle stage and the later stage of fermentation, wherein the fermentation temperature is higher in the early stage of fermentation, the fermentation temperature is lower in the middle stage of fermentation, and the fermentation temperature is increased in the later stage of fermentation;
(c) after fermentation, adding coconut oil into the fermentation liquor obtained in the step (b), mixing and stirring uniformly to obtain a fermentation mixed liquor, concentrating the mixed liquor, drying the thalli, detecting and packaging to obtain the blue algae purifying agent.
2. The method of claim 1, wherein:
the inoculation amounts of the Bacillus cereus and Bacillus subtilis liquid in the step (b) are 3-8% of the weight of the liquid fermentation medium;
the fermentation culture conditions are as follows: the initial pH value is 7.0-7.5, the rotating speed is 150-180 r/min, the tank pressure is 0.04-0.06MPa, the fermentation time is 24-28 h, the temperature is controlled at 35-39 ℃ for 1-6 h, the temperature is controlled at 28-32 ℃ for 6-20 h, and the temperature is controlled at 33-37 ℃ for 20h till the end of fermentation.
3. The method of claim 1, wherein:
the liquid fermentation medium in the step (b) consists of the following components: the fertilizer comprises, by weight, 4-6% of cyanobacteria powder, 0.2-0.5% of ammonium sulfate, 0.1-0.2% of dipotassium hydrogen phosphate, 0.05% of magnesium sulfate, 0.05% of ferrous sulfate, 0.01% of manganese sulfate, 0.01% of sodium chloride and the balance of water. After the liquid fermentation medium is prepared according to the formula, the pH value of the liquid fermentation medium is adjusted to 7.0-7.5.
4. The method of claim 1, wherein:
and (c) after the fermentation in the step (c) is finished, adding coconut oil into the fermentation liquor in an amount which is 8-12% of the volume of the fermentation liquor, and mixing and stirring for 1-2 hours.
5. The method of claim 1, wherein:
the method for concentrating the mixed solution in the step (c) is to collect thalli after plate-frame filtration; the method for drying the thalli is a conventional physical drying method.
6. The method of claim 2, wherein:
the inoculation amounts of the Bacillus cereus and the Bacillus subtilis in the step (b) are both 5% of the weight of the liquid fermentation medium;
the fermentation culture conditions are as follows: the initial pH value is 7.5, the rotating speed is 180r/min, the tank pressure is 0.04-0.06MPa, the fermentation time is 26h, the temperature is controlled at 37 ℃ for 1-6 h, the temperature is controlled at 30 ℃ for 6-20 h, and the temperature is controlled at 35 ℃ after 20h and fermentation is finished.
7. A method according to claim 3, characterized by:
the liquid fermentation medium in the step (b) consists of the following components: the blue algae powder comprises, by weight, 5% of blue algae powder, 0.3% of ammonium sulfate, 0.2% of dipotassium phosphate, 0.05% of magnesium sulfate, 0.05% of ferrous sulfate, 0.01% of manganese sulfate, 0.01% of sodium chloride and the balance of water. After the liquid fermentation medium is prepared according to the formula, the pH value is adjusted to 7.5.
8. The method of claim 4, wherein:
after the fermentation in the step (c) is finished, the volume of the coconut oil added into the fermentation liquid is 10% of the volume of the fermentation liquid, and the mixing and stirring time is 2 hours.
9. A cyanobacteria scavenger obtained by the method according to any one of claims 1 to 8.
10. The use of the cyanobacteria scavenger according to claim 9 in the improvement of the aquatic environment of aquaculture water.
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Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101211679B1 (en) * 2011-06-02 2012-12-12 (주)지앤브이 Method for producing the decomposing agent of microorganism and decomposing agent of microorganism made thereby
CN103160453A (en) * 2013-02-26 2013-06-19 武汉施瑞福生物技术有限公司 Preparation method of microecologics for nitrite degradation, microecologics and use thereof
CN103193329A (en) * 2013-04-15 2013-07-10 中国水产科学研究院淡水渔业研究中心 Biological agent for controlling excessive propagation of blue-green algae of aquaculture water and preparation method of biological agent
KR20140077645A (en) * 2012-12-14 2014-06-24 주식회사 한국아쿠아서비스 Microbial agent formulations for improving water quality and sediment quality
CN105087426A (en) * 2015-06-25 2015-11-25 中国水产科学研究院南海水产研究所 Industrial liquid-solid combined fermentation method for bacillus cereus preparation for dissolving oscillatoria in aquaculture pond
WO2017081105A1 (en) * 2015-11-09 2017-05-18 Institute Of Genetics And Developmental Biology, Chinese Academy Of Sciences Bacillus strains and agents with beneficial properties
CN106967655A (en) * 2017-05-24 2017-07-21 武汉绿富农业生物工程技术股份有限公司 A kind of algae phase modifier and preparation method and application
CN107265657A (en) * 2017-06-20 2017-10-20 森信必高投资有限公司 Active microorganism combination biological agent is used for the method for administering blue-green algae
CN108277179A (en) * 2018-03-07 2018-07-13 武汉光谷创赢生物技术开发有限公司 A kind of high density fermentation and spray drying process of bacillus subtilis
CN109593681A (en) * 2018-12-28 2019-04-09 天津坤禾生物科技集团股份有限公司 A kind of composite bacteria agent and its preparation method and application for preventing and treating aquaculture cyanobacterial bloom

Patent Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR101211679B1 (en) * 2011-06-02 2012-12-12 (주)지앤브이 Method for producing the decomposing agent of microorganism and decomposing agent of microorganism made thereby
KR20140077645A (en) * 2012-12-14 2014-06-24 주식회사 한국아쿠아서비스 Microbial agent formulations for improving water quality and sediment quality
CN103160453A (en) * 2013-02-26 2013-06-19 武汉施瑞福生物技术有限公司 Preparation method of microecologics for nitrite degradation, microecologics and use thereof
CN103193329A (en) * 2013-04-15 2013-07-10 中国水产科学研究院淡水渔业研究中心 Biological agent for controlling excessive propagation of blue-green algae of aquaculture water and preparation method of biological agent
CN105087426A (en) * 2015-06-25 2015-11-25 中国水产科学研究院南海水产研究所 Industrial liquid-solid combined fermentation method for bacillus cereus preparation for dissolving oscillatoria in aquaculture pond
WO2017081105A1 (en) * 2015-11-09 2017-05-18 Institute Of Genetics And Developmental Biology, Chinese Academy Of Sciences Bacillus strains and agents with beneficial properties
CN106967655A (en) * 2017-05-24 2017-07-21 武汉绿富农业生物工程技术股份有限公司 A kind of algae phase modifier and preparation method and application
CN107265657A (en) * 2017-06-20 2017-10-20 森信必高投资有限公司 Active microorganism combination biological agent is used for the method for administering blue-green algae
CN108277179A (en) * 2018-03-07 2018-07-13 武汉光谷创赢生物技术开发有限公司 A kind of high density fermentation and spray drying process of bacillus subtilis
CN109593681A (en) * 2018-12-28 2019-04-09 天津坤禾生物科技集团股份有限公司 A kind of composite bacteria agent and its preparation method and application for preventing and treating aquaculture cyanobacterial bloom

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
G. PRABAKARAN等: "Coconut water as a cheap source for the production of δ endotoxin of Bacillus thuringiensis var. israelensis, a mosquito control agent", ACTA TROPICA, vol. 105, no. 1, pages 35 - 38, XP022392559, DOI: 10.1016/j.actatropica.2007.09.002 *
XIANGDONG BI等: "Effects of Bacillus subtilis on the growth, colony maintenance, and attached bacterial community composition of colonial cyanobacteria", ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH, vol. 26, pages 14977 - 14987 *
张克强等: "芽孢杆菌菌剂在水产养殖中的应用初探", 海洋科学, vol. 30, no. 09, pages 88 - 91 *

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