CN114958638B - Composite microbial agent for reducing microcystin content in aquatic products and application method thereof - Google Patents
Composite microbial agent for reducing microcystin content in aquatic products and application method thereof Download PDFInfo
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- CN114958638B CN114958638B CN202011283822.7A CN202011283822A CN114958638B CN 114958638 B CN114958638 B CN 114958638B CN 202011283822 A CN202011283822 A CN 202011283822A CN 114958638 B CN114958638 B CN 114958638B
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- 239000002131 composite material Substances 0.000 title claims abstract description 53
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- 108010067094 microcystin Proteins 0.000 title claims abstract description 25
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- GLFNIEUTAYBVOC-UHFFFAOYSA-L Manganese chloride Chemical compound Cl[Mn]Cl GLFNIEUTAYBVOC-UHFFFAOYSA-L 0.000 description 1
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- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
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- ZYZCGGRZINLQBL-GWRQVWKTSA-N microcystin-LR Chemical compound C([C@H](OC)[C@@H](C)\C=C(/C)\C=C\[C@H]1[C@@H](C(=O)N[C@H](CCC(=O)N(C)C(=C)C(=O)N[C@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H]([C@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N1)C(O)=O)C(O)=O)C)C1=CC=CC=C1 ZYZCGGRZINLQBL-GWRQVWKTSA-N 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
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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
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/80—Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
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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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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Abstract
The invention relates to a composite microbial agent for reducing the content of microcystin in aquatic products, which comprises Brevundimonas deficiency and Pseudomonas stutzeri, wherein the composite microbial agent is mixed bacterial liquid formed by mixing Brevundimonas deficiency bacterial liquid and Pseudomonas stutzeri bacterial liquid; the Brevundimonas deficiency is Brevundimonas deficiency (Brevundimonas diminuta) Z1, and the preservation number is CGMCC No. 20009; the pseudomonas stutzeri is pseudomonas stutzeri (Pseudomonas stutzeri) ZJ4, and the preservation number is CGMCC No. 20910. The invention obviously improves the degradation capability of the microcystins in the fish intestinal tracts, reduces the absorption amount of the microcystins through the intestinal tracts, and reduces the accumulation amount of the microcystins in the fish bodies; promote denitrification and removal of ammonia nitrogen and nitrate nitrogen in water and growth of chlorella and other green algae, reduce biomass of toxic blue algae, and form virtuous circle of microcystin control in culture environment.
Description
Technical Field
The invention relates to a composite microbial agent for reducing the content of microcystin in aquatic products and a use method thereof, belonging to the technical field of environmental protection.
Background
Along with the development of large-scale and high-density fishery cultivation in China, a large amount of feed which is not eaten by fishes and feces discharged by the fishes in the cultivation process make the problem of eutrophication of pond water quality more serious. Blue algae is used as one of primary producers of water, the propagation speed is too high, the blue algae is easy to become dominant algae, water bloom is formed in a culture pond, the blue algae can be propagated quickly after being treated by chemical agents or algae-dissolving bacteria, and hidden danger cannot be eradicated. The microcystin is an extremely strong liver tumor promoter generated by dominant species of common cyanobacteria bloom in China, can accumulate in aquatic animals including fish, generates toxic and side effects, and is transmitted through food chain media, so that the risk of harm to human health is generated. The existing solution method mainly promotes the conversion of the microcystin in the fish body and the discharge of the microcystin outside by adding antioxidants such as glutathione, vitamin C, ligustrazine (traditional Chinese medicine), but the added antioxidants are easy to be interfered by oxidation factors in the environment to cause a great deal of loss, so that the effect is greatly reduced, the effect can be maintained only by increasing the adding amount, and the cultivation cost is further increased. The other methods are to add probiotics such as lactobacillus to promote the immune function of fish, but the bacteria have proper growth pH value and slightly acidic property, are not suitable for the intestinal microenvironment of neutral and slightly alkaline of main domestic fishes such as grass carp, silver carp and the like, and are difficult to effectively colonize in the intestinal tract of fish and exert the optimal effect.
With the intensive research, it is found that other bacteria also have the ability to degrade algae toxins, and chinese patent publication No. CN110791449a provides a microcystin degrading bacteria and application thereof, and the patent discloses application of microbacterium or its fermentation product in degrading microcystin, which essentially comprises extracting and concentrating protease substances with algae toxin degrading function in cells of the bacteria after cell disruption treatment, so that the extract must be used with protective agent to reduce denaturation and inactivation due to influence of protein accepting and chemical factors, so as to adapt to complex environment in fish digestive tract. However, ubiquitous microorganisms still develop stress on proteolytic enzymes, including microorganisms in the fish gut, resulting in poor degradation of microcystins.
Disclosure of Invention
The technical problems to be solved by the invention are as follows: aiming at the problems existing in the prior art, a composite microbial inoculum for reducing the content of microcystin in aquatic products and a use method thereof are provided.
In order to achieve the above purpose, the technical scheme of the invention is as follows:
A composite microbial agent for reducing the microcystin content in aquatic products comprises Brevundimonas deficiency and Pseudomonas stutzeri, wherein the composite microbial agent is a mixed bacterial liquid formed by mixing the Brevundimonas deficiency bacterial liquid and the Pseudomonas stutzeri bacterial liquid; the Brevundimonas deficiency is Brevundimonas deficiency (Brevundimonas diminuta) Z1, and the preservation number is CGMCC No. 20009; the pseudomonas stutzeri is pseudomonas stutzeri (Pseudomonas stutzeri) ZJ4, and the preservation number is CGMCC No. 20910.
The invention separates bacteria for efficiently degrading algae toxin from fish body or fish feces, and the bacteria separated from the fish body or the fish feces have stronger field planting and capability of adapting to the environment of the fish digestive tract. Because colonization is host-specific, e.g., bacteria from the human body, it is not necessarily possible to colonize the animal body; colonization also has strain specificity, and some of the same genus of bacteria may be, but some may not. The two bacteria contained in the composite bacterial agent disclosed by the invention can adapt to the internal environment of the fish body.
In the technical scheme, the composite microbial inoculum is a mixed microbial inoculum formed by mixing 10 8～1011 CFU/mL of defective shortwave monad liquid and 10 8～1011 CFU/mL of pseudomonas stutzeri liquid according to the volume ratio of 1-2:10-1.
Further, the compound bacteria are prepared by respectively culturing Brevundimonas imperfecti and Pseudomonas stutzeri, concentrating to 10 11 CFU/mL, and mixing at a volume ratio of 1:2.
The composite microbial inoculum also comprises a synergistic auxiliary agent, wherein the synergistic auxiliary agent accounts for 2-8% of the weight of the mixed microbial inoculum.
Further, the synergistic agent is at least one of glucose, molasses, sucrose, galactose and flour.
The invention screens pseudomonas stutzeri and shortwave pseudomonas defectives from fish bodies or fish feces, can grow in a culture medium which takes MC-LR (microcystin LR) as a sole carbon source and nitrogen source, and can degrade microcystin. And the synergistic additives such as glucose, sucrose, molasses and the like are added in the carbon source, so that the degradation efficiency of the algae toxins can be further improved.
Still further, the co-agent comprises 2% glucose by weight of the mixed liquor and 6% flour by weight of the mixed liquor.
The invention also provides a use method of the composite microbial inoculum, which comprises the following steps:
Firstly, placing shortwave pseudomonas defectives and pseudomonas stutzeri in 500-5000 mL of LB liquid culture medium respectively, culturing for 12-60 hours at 20-40 ℃, concentrating the cultured bacterial liquid to 10 8～1011 CFU/mL respectively, and mixing the concentrated bacterial liquid to prepare mixed bacterial liquid in a volume ratio of 1-2:10-1;
Secondly, adding 2-8% of synergistic auxiliary agent into the mixed bacterial liquid according to the weight percentage of the mixed bacterial liquid, and stirring to dissolve the synergistic auxiliary agent into the mixed bacterial liquid to form a composite bacterial agent;
and thirdly, directly spraying the composite microbial inoculum on the surface of the commercially available puffed compound feed for fish, and air-drying the composite microbial inoculum for use.
Further, the synergistic agent is at least one of glucose, molasses, sucrose, galactose and flour.
According to the invention, through feeding of the bacteria-containing feed, the relative abundance of specific bacteria in fish intestinal tracts in the colonization flora can be improved, so that the degradation capability of microcystins in fish intestinal tracts is improved, the absorption amount of the microcystins through intestinal tracts is reduced, and the accumulation amount of the microcystins in fish bodies is reduced. Meanwhile, the bacteria-containing feed which is not eaten by fishes and the feces discharged by the fishes can accelerate the decomposition and conversion of the feed residues under the action of the composite microbial inoculum in the feed, so that the bacteria are amplified in the water body; the screened specific strain has the effect of promoting the growth of the green algae, so that the nutrition competition of the green algae and the blue algae in the culture water body is promoted, the biomass of the blue algae is reduced, the amount of blue algae eaten by fish is reduced from the source, and the virtuous circle of culture is formed.
The application method of the composite microbial inoculum specifically comprises the following steps:
Firstly, respectively placing shortwave pseudomonas defectives and pseudomonas stutzeri in 1000mL of LB liquid culture medium, culturing for 36 hours at 28 ℃, respectively concentrating the cultured bacterial liquid to 10 11 CFU/mL, and mixing the concentrated bacterial liquid in a volume ratio of 1:2 to prepare mixed bacterial liquid;
Secondly, adding 2% of glucose and 6% of flour into the mixed bacterial liquid according to the weight percentage of the mixed bacterial liquid, and stirring to dissolve the glucose and the flour into the mixed bacterial liquid to form a composite bacterial agent;
and thirdly, directly spraying the composite microbial inoculum on the surface of the commercially available puffed compound feed for fish, and air-drying the composite microbial inoculum for use.
Further, the viable count of the shortwave monad defect in the composite microbial inoculum is 0.5X10 7CFU/g～2×107 CFU/g, and the viable count of the pseudomonas stutzeri is 1× 7CFU/g～4×107 CFU/g.
The invention has the advantages that the degradation capability of the microcystins in the fish intestinal canal is obviously improved by the composite microbial inoculum, the absorption amount of the microcystins through the intestinal canal is reduced, and the accumulation amount of the microcystins in the fish body is further reduced; meanwhile, a large amount of feeds which are not eaten by fishes promote the growth of scattered microbial inoculum, so that the denitrification removal of ammonia nitrogen and nitrate nitrogen in the water body can be promoted, the growth of green algae such as chlorella and the like can be promoted, the nutrition competition of the green algae and blue algae in the culture water body can be finally promoted, the biomass of the toxic blue algae can be reduced, and the virtuous circle of microcystin control in the culture environment can be formed.
Detailed Description
The present invention will be described in further detail with reference to examples. The invention is not limited to the examples given.
The chemical reagents and materials used in the invention are all commercially available.
Example 1
The embodiment provides a composite microbial inoculum for reducing the microcystin content in aquatic products, which comprises mixed bacterial liquid formed by mixing 10 11 CFU/mL of shortwave monad liquid and 10 11 CFU/mL of pseudomonas stutzeri liquid according to the volume ratio of 1:2. Screening short wave monad Brevundimonas diminutaZ from silver carp excrement, wherein the preservation number is CGMCC No. 20209, the preservation unit is China general microbiological culture Collection center, the address is North Star Xway No. 1, 3 of the Korean area of Beijing, and the preservation date is 2020, 10 months and 19 days; and Pseudomonas stutzeri Pseudomonas stutzeriZJ4 with the preservation number of CGMCC No. 2091, the preservation unit of China general microbiological culture Collection center, the address of the China general microbiological culture Collection center, the No. 3 of Xili No. 1 in the Guangxi district North Star of Beijing, and the preservation date of the China general microbiological culture Collection center of 2020, 10 months and 19 days. Both strains can be grown in a medium with MC-LR as the sole carbon source and nitrogen source to degrade microcystin. By adding the two bacteria into the fish feed, the relative abundance of the two bacteria in the intestinal tract of the silver carp can be obviously improved, which indicates that the silver carp can be better planted in the intestinal tract of the silver carp. Both bacteria are facultative heterotrophic bacteria, feed which is not eaten by fishes and excrement discharged by fishes can be just used as a substrate for culturing the same, pseudomonas stutzeri Pseudomonas stutzeriZJ4 can promote denitrification removal of ammoniacal nitrogen and nitrate nitrogen in a water body, brevundimonas defecate Brevundimonas diminuta Z1 can promote growth of green algae such as chlorella and the like, promote nutrition competition of green algae and blue algae in a culture water body, and reduce biomass of the blue algae.
The mixed bacterial liquid can be added with a synergistic auxiliary agent to improve the degradation efficiency of the bacterial agent on the microcystin, wherein the synergistic auxiliary agent accounts for 2-8% of the weight of the mixed bacterial liquid, and the synergistic auxiliary agent is at least one of glucose, molasses, sucrose, galactose and flour.
The shortwave monad Brevundimonas diminutaZ and the pseudomonas stutzeri Pseudomonas stutzeriZJ4 are respectively inoculated into culture mediums containing microcystins with different concentrations (the culture mediums are taken as basic culture mediums, microcystins with different concentrations are added as unique carbon sources and nitrogen sources, the basic culture medium formula is MgSO4·7H2O 1g/L, KH2PO40.5g/L, K2HPO44.0g/L, NaCl 1.0 g/L,CaCl220 mg/L,FeSO45 mg/L,ZnCl25 mg/L, MnCl2·4H2O 5mg/L,CuCl20.5mg/L）, experiments show that the 48h degradation efficiency of the shortwave monad Brevundimonas diminuta Z1 to the microcystins MC-LR with the concentration of 0.45-8 mg/L is better and reaches more than 64 percent, while the 48h degradation efficiency of the pseudomonas stutzeri Pseudomonas stutzeri ZJ is particularly less different from that of the microcystins MC-LR with the concentration of <0.6 mg/L and reaches more than 43 percent, and the advantages of the microcystins degrading bacteria on the microcystins with different concentrations can be fully exerted.
In addition, the synergistic auxiliary agents such as glucose, flour, molasses, sucrose, galactose and the like are added into the culture medium, so that the degradation efficiency of the two bacteria on the microcystin MC-LR can be further obviously improved, the degradation efficiency of the defect Brevundimonas Brevundimonas diminutaZ1 on the microcystin MC-LR is improved by 12.8%, and the degradation efficiency of the Pseudomonas stutzeri Pseudomonas stutzeriZJ4 on the microcystin MC-LR is improved by 18.3%.
Example 2
The embodiment also provides a use method of the composite microbial inoculum, which specifically comprises the following steps:
Firstly, respectively placing shortwave monad Brevundimonas diminuta Z and pseudomonas stutzeri Pseudomonas stutzeriZJ4 in 500mL of LB liquid culture medium, culturing for 18 hours at 35 ℃, respectively concentrating the cultured bacterial liquid to 10 11 CFU/mL, and mixing the concentrated bacterial liquid in a volume ratio of 1:2 to prepare mixed bacterial liquid;
Secondly, adding 2% of glucose and 6% of flour into the mixed bacterial liquid according to the weight percentage of the mixed bacterial liquid, and stirring to dissolve the glucose and the flour into the mixed bacterial liquid to form a composite bacterial agent; the viable count of the shortwave monad Brevundimonas diminutaZ to be defective in the composite bacterial agent is 0.5X10 7CFU/g～2×107 CFU/g, and the viable count of the pseudomonas stutzeri Pseudomonas stutzeriZJ to be 1X 10 7CFU/g～4×107 CFU/g;
and thirdly, spraying the composite microbial inoculum on the surface of the commercially available puffed compound feed for fish, and directly putting the fish into a culture pond for use after air drying.
Example 3
Silver carp culture is used as an experiment, the silver carp throwing density in a culture pond is 1.6kg/m 2, the daily feeding amount is 2.5% of the weight, and the concentration of the blue algae with toxin production and microcystis aeruginosa is kept at 1X 10 6～5×106/ml. After 42 days, the relative abundance of Brevibacterium deficiency Brevundimonas diminuta and Pseudomonas stutzeri Pseudomonas stutzeri in the intestinal flora of the silver carp in the culture pond of the bacterial-containing feed of the example 2 is increased, wherein Brevundimonas deficiency Brevundimonas diminuta is increased from 0.057% to 4.46%, pseudomonas stutzeri Pseudomonas stutzeri is increased from 0.032% to 1.74% and the total antioxidant capacity in the liver of the silver carp is remarkably improved from 12.78U/mg to 16.13U/mg, 26.21% and the MDA content of the liver of the silver carp is remarkably reduced from 9.4nmol/mg to 6.1nmol/mg, 35.11% and the algal toxin content in the muscle of the silver carp is remarkably reduced from 61.64 ng/g to 47.37 ng/g. Experimental results show that the compound bacterial agent can promote the improvement of the abundance of microcystins degrading microorganisms in fish intestinal tracts, so that the degradation capability of the microcystins in the fish intestinal tracts is improved, the absorption amount of the microcystins through the intestinal tracts is reduced, and the accumulation amount of the microcystins in fish bodies is reduced.
Example 4
The chlorella Chlorella ellipsoidea and the microcystis aeruginosa are co-cultured, the initial concentration of the chlorella is 3 multiplied by 10 5～4×105/ml, then the short-wave-strain is added for Brevundimonas diminutaZ, 7.3 multiplied by 10 4/ml, the chlorella is observed after 42 days to be used as a control without the short-wave-strain, the chlorella concentration in the control is 4.2 multiplied by 10 6/ml, the chlorella producing the toxic is 1.7 multiplied by 10 7/ml, the chlorella concentration in the treatment of the short-wave-strain adding Brevundimonas diminutaZ is 8.1 multiplied by 10 6/ml, the chlorella producing the toxic is 7.6 multiplied by 10 6/ml, and the short-wave-strain adding Brevundimonas diminutaZ promotes the biomass improvement of the chlorella and reduces the biomass of the toxic blue-green algae. Experimental results show that Brevundimonas defective Brevundimonas diminutaZ has a promoting effect on the growth of green alga Chlorella Chlorella ellipsoidea.
Example 5
The embodiment also provides a use method of the composite microbial inoculum, which specifically comprises the following steps:
Firstly, respectively placing the shortwave monad Brevundimonas diminuta Z and the pseudomonas stutzeri Pseudomonas stutzeriZJ in 1000mL of LB liquid culture medium, culturing for 60 hours at 28 ℃, respectively concentrating the cultured bacterial liquid to 10 8 CFU/mL, and mixing the concentrated bacterial liquid in a volume ratio of 1:2 to prepare mixed bacterial liquid;
Secondly, adding molasses and galactose accounting for 2 percent of the weight percent of the mixed bacterial liquid into the mixed bacterial liquid, and stirring to dissolve the molasses and galactose in the mixed bacterial liquid to form a composite bacterial agent; the viable count of the shortwave monad Brevundimonas diminutaZ to be defective in the composite bacterial agent is 0.5X10 7CFU/g～1×107 CFU/g, and the viable count of the pseudomonas stutzeri Pseudomonas stutzeriZJ to be 1X 10 7CFU/g～1.5×107 CFU/g;
thirdly, spraying the composite microbial inoculum on the surface of the commercial puffed compound feed for the fish, and directly putting the fish into a culture pond for use after air drying, wherein the daily feeding amount is generally 2.5% of the weight of the fish.
Example 6
The embodiment also provides a use method of the composite microbial inoculum, which specifically comprises the following steps:
Firstly, respectively placing shortwave monad Brevundimonas diminuta Z and pseudomonas stutzeri Pseudomonas stutzeriZJ4 in 5000mL of LB liquid culture medium, culturing at 40 ℃ for 36 hours, respectively concentrating the cultured bacterial liquid to 10 9 CFU/mL, and mixing the concentrated bacterial liquid in a volume ratio of 1:2 to prepare mixed bacterial liquid;
Step two, adding sucrose accounting for 2 percent of the mixed bacterial liquid in percentage by weight into the mixed bacterial liquid, and stirring to dissolve the sucrose in the mixed bacterial liquid to form a composite bacterial agent; the viable count of the shortwave monad Brevundimonas diminutaZ to be defective in the composite bacterial agent is 0.5X10 7CFU/g～1×107 CFU/g, and the viable count of the pseudomonas stutzeri Pseudomonas stutzeriZJ to be 1X 10 7CFU/g～1.5×107 CFU/g;
thirdly, spraying the composite microbial inoculum on the surface of the commercial puffed compound feed for the fish, and directly putting the fish into a culture pond for use after air drying, wherein the daily feeding amount is generally 2.5% of the weight of the fish.
In addition to the embodiments described above, other embodiments of the invention are possible. All technical schemes formed by equivalent substitution or equivalent transformation fall within the protection scope of the invention.
Claims (8)
1. A composite microbial agent for reducing the content of microcystin in aquatic products is characterized by comprising Brevundimonas imperfecti and Pseudomonas stutzeri; the Brevundimonas deficiency is Brevundimonas deficiency (Brevundimonas diminuta) Z1, and the preservation number is CGMCC No. 20009; the pseudomonas stutzeri is pseudomonas stutzeri (Pseudomonas stutzeri) ZJ4, and the preservation number is CGMCC No. 20910; the composite bacterial agent is a mixed bacterial liquid formed by mixing 10 8～1011 CFU/mL of shortwave monad liquid and 10 8～1011 CFU/mL of pseudomonas stutzeri liquid according to a volume ratio of 1:2.
2. The composite microbial agent for reducing the microcystin content in aquatic products according to claim 1, wherein the composite microbial agent is prepared by respectively culturing Brevundimonas imperfecti and Pseudomonas stutzeri, concentrating to 10 11 CFU/mL, and mixing the concentrated bacterial solutions in a volume ratio of 1:2.
3. The composite microbial agent for reducing the microcystin content in aquatic products according to claim 1, wherein the composite microbial agent further comprises a synergistic agent, and the synergistic agent accounts for 2% -8% of the weight of the mixed microbial liquid.
4. The composite microbial agent for reducing the microcystin content in aquatic products according to claim 3, wherein the synergistic agent is at least one of glucose, molasses, sucrose, galactose and flour.
5. The composite microbial inoculant for reducing the microcystin content in an aquatic product according to claim 4, wherein the synergistic agent comprises 2% of glucose by weight of the mixed microbial inoculant and 6% of flour by weight of the mixed microbial inoculant.
6. The method of using a composite microbial agent according to any one of claims 1 to 5, comprising the steps of:
firstly, placing shortwave pseudomonas defectives and pseudomonas stutzeri in 500-5000 mL of LB liquid culture medium respectively, culturing for 12-60 hours at 20-40 ℃, concentrating the cultured bacterial liquid to 10 8～1011 CFU/mL respectively, and mixing the concentrated bacterial liquid in a volume ratio of 1:2 to prepare mixed bacterial liquid;
Secondly, adding 2-8% of synergistic auxiliary agent into the mixed bacterial liquid according to the weight percentage of the mixed bacterial liquid, and stirring to dissolve the synergistic auxiliary agent into the mixed bacterial liquid to form a composite bacterial agent;
and thirdly, directly spraying the composite microbial inoculum on the surface of the commercially available puffed compound feed for fish, and air-drying the composite microbial inoculum for use.
7. The method of using the composite microbial inoculant of claim 6, comprising the steps of:
firstly, respectively placing shortwave pseudomonas defectives and pseudomonas stutzeri in 1000mL of LB liquid culture medium, culturing for 36 hours at 28 ℃, respectively concentrating the cultured bacterial liquid to 10 11 CFU/mL, and mixing the concentrated bacterial liquid in a volume ratio of 1:2 to prepare mixed bacterial liquid;
Secondly, adding 2% of glucose and 6% of flour into the mixed bacterial liquid according to the weight percentage of the mixed bacterial liquid, and stirring to dissolve the glucose and the flour into the mixed bacterial liquid to form a composite bacterial agent;
and thirdly, directly spraying the composite microbial inoculum on the surface of the commercially available puffed compound feed for fish, and air-drying the composite microbial inoculum for use.
8. The method for using the composite microbial inoculant according to claim 7, wherein the viable count of the shortwave monad defect in the composite microbial inoculant is 0.5X10 7CFU/g～2×107 CFU/g, and the viable count of the pseudomonas stutzeri is 1X 10 7CFU/g～4×107 CFU/g.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH05112417A (en) * | 1991-02-28 | 1993-05-07 | Mercian Corp | Red tide-controlling feed for fish |
| CN105567589A (en) * | 2015-12-29 | 2016-05-11 | 深圳市铁汉生态环境股份有限公司 | Brevundimonas diminuta and application thereof |
| CN106635857A (en) * | 2015-11-04 | 2017-05-10 | 中国石油化工股份有限公司 | Pseudomonas stutzeri and culture application thereof |
| CN107739722A (en) * | 2017-09-15 | 2018-02-27 | 常州大学 | A kind of method that algal toxin degradation bacterium is screened in the turbulent waves fish guts from Taihu Lake |
| CN107986456A (en) * | 2017-12-14 | 2018-05-04 | 泰州永达生物科技有限公司 | A kind of prevention and suppressing method applied to water body cyanobacteria |
| CN111471609A (en) * | 2019-12-26 | 2020-07-31 | 中国科学院水生生物研究所 | Pseudomonas with algae-lysing activity and application thereof |
-
2020
- 2020-11-17 CN CN202011283822.7A patent/CN114958638B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH05112417A (en) * | 1991-02-28 | 1993-05-07 | Mercian Corp | Red tide-controlling feed for fish |
| CN106635857A (en) * | 2015-11-04 | 2017-05-10 | 中国石油化工股份有限公司 | Pseudomonas stutzeri and culture application thereof |
| CN105567589A (en) * | 2015-12-29 | 2016-05-11 | 深圳市铁汉生态环境股份有限公司 | Brevundimonas diminuta and application thereof |
| CN107739722A (en) * | 2017-09-15 | 2018-02-27 | 常州大学 | A kind of method that algal toxin degradation bacterium is screened in the turbulent waves fish guts from Taihu Lake |
| CN107986456A (en) * | 2017-12-14 | 2018-05-04 | 泰州永达生物科技有限公司 | A kind of prevention and suppressing method applied to water body cyanobacteria |
| CN111471609A (en) * | 2019-12-26 | 2020-07-31 | 中国科学院水生生物研究所 | Pseudomonas with algae-lysing activity and application thereof |
Non-Patent Citations (5)
| Title |
|---|
| 一株降解微囊藻毒素的缺陷短波单胞菌的分离鉴定及降解特性研究;谷青;赵琪;史全良;;环境科学学报(第01期);第201页摘要 * |
| 微囊藻毒素降解菌EMB分离鉴定及其降解特性;尹玉芬;胡梁斌;周威;杨静东;周益军;石志琦;;江苏农业学报(第03期);第631-636页 * |
| 斯氏假单胞菌对草鱼养殖水体水质的调控作用;郑佳佳;张小平;胡彩虹;李卫芬;;渔业现代化(第03期);第5-9页 * |
| 溶藻细菌及其测定评价方法的研究进展;赵传鹏, 浦跃朴, 尹立红;东南大学学报(医学版)(第03期);第202-206页 * |
| 赵冬志等.中国典型海域赤潮灾害发生规律.北京：海洋出版社,2010,第311页. * |
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