CN113564087B - Biological algicide and preparation method and application thereof - Google Patents
Biological algicide and preparation method and application thereof Download PDFInfo
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- 239000003619 algicide Substances 0.000 title claims abstract description 67
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
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- 241000195493 Cryptophyta Species 0.000 claims abstract description 51
- 229920001661 Chitosan Polymers 0.000 claims abstract description 48
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- 238000000034 method Methods 0.000 claims abstract description 20
- 239000007788 liquid Substances 0.000 claims abstract description 19
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 18
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- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 4
- 229960000583 acetic acid Drugs 0.000 claims description 4
- LFVGISIMTYGQHF-UHFFFAOYSA-N ammonium dihydrogen phosphate Chemical compound [NH4+].OP(O)([O-])=O LFVGISIMTYGQHF-UHFFFAOYSA-N 0.000 claims description 4
- 229910000387 ammonium dihydrogen phosphate Inorganic materials 0.000 claims description 4
- 235000005822 corn Nutrition 0.000 claims description 4
- 239000012362 glacial acetic acid Substances 0.000 claims description 4
- 239000008103 glucose Substances 0.000 claims description 4
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 claims description 4
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 claims description 4
- 235000019837 monoammonium phosphate Nutrition 0.000 claims description 4
- 239000006012 monoammonium phosphate Substances 0.000 claims description 4
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 4
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 4
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 4
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
- 239000007787 solid Substances 0.000 claims description 4
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- 235000019698 starch Nutrition 0.000 claims description 4
- 241001052560 Thallis Species 0.000 claims description 3
- 239000008367 deionised water Substances 0.000 claims description 3
- 229910021641 deionized water Inorganic materials 0.000 claims description 3
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- 230000000694 effects Effects 0.000 description 7
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- 239000013543 active substance Substances 0.000 description 4
- 229910000365 copper sulfate Inorganic materials 0.000 description 4
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 4
- 239000002028 Biomass Substances 0.000 description 3
- 229910021380 Manganese Chloride Inorganic materials 0.000 description 3
- GLFNIEUTAYBVOC-UHFFFAOYSA-L Manganese chloride Chemical compound Cl[Mn]Cl GLFNIEUTAYBVOC-UHFFFAOYSA-L 0.000 description 3
- 241000519590 Pseudoalteromonas Species 0.000 description 3
- 238000012851 eutrophication Methods 0.000 description 3
- SURQXAFEQWPFPV-UHFFFAOYSA-L iron(2+) sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Fe+2].[O-]S([O-])(=O)=O SURQXAFEQWPFPV-UHFFFAOYSA-L 0.000 description 3
- 239000011565 manganese chloride Substances 0.000 description 3
- 235000002867 manganese chloride Nutrition 0.000 description 3
- 229940099607 manganese chloride Drugs 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
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- 239000011684 sodium molybdate Substances 0.000 description 3
- 235000015393 sodium molybdate Nutrition 0.000 description 3
- TVXXNOYZHKPKGW-UHFFFAOYSA-N sodium molybdate (anhydrous) Chemical compound [Na+].[Na+].[O-][Mo]([O-])(=O)=O TVXXNOYZHKPKGW-UHFFFAOYSA-N 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000011573 trace mineral Substances 0.000 description 3
- 235000013619 trace mineral Nutrition 0.000 description 3
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 3
- 229910000368 zinc sulfate Inorganic materials 0.000 description 3
- 229960001763 zinc sulfate Drugs 0.000 description 3
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 241000863422 Myxococcus xanthus Species 0.000 description 2
- 241000109965 Pseudoalteromonas marina Species 0.000 description 2
- 238000010170 biological method Methods 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 238000000053 physical method Methods 0.000 description 2
- 230000003248 secreting effect Effects 0.000 description 2
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- 241000894006 Bacteria Species 0.000 description 1
- 241001474374 Blennius Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000192710 Microcystis aeruginosa Species 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 239000005708 Sodium hypochlorite Substances 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 239000013043 chemical agent Substances 0.000 description 1
- 239000004927 clay Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000002360 explosive Substances 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 230000000737 periodic effect Effects 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229920000867 polyelectrolyte Polymers 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000004094 surface-active agent Substances 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
-
- 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
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- Biotechnology (AREA)
- Zoology (AREA)
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Abstract
The invention relates to the technical field of water treatment, in particular to a biological algicide and a preparation method and application thereof. The biological algicide is obtained by modifying chitosan after fermenting and culturing a composite microbial inoculum, wherein the composite microbial inoculum comprises actinomycetes, bacillus, alteromonas and myxobacteria. The preparation method of the biological algicide comprises the following steps: (1) Fermenting and culturing the composite microbial inoculum to obtain composite bacterial liquid; (2) Mixing the compound bacterial liquid in the step (1) with the chitosan solution in proportion, and adopting the chitosan modified compound bacterial agent to obtain the biological algicide. The biological algicide is obtained by modifying chitosan after fermenting and culturing the composite microbial inoculum, has strong environmental change tolerance in the actual use process, has the advantages of simple operation, low cost and no secondary pollution when being applied to algae removal in an algae outbreak water body, and has high algae removal efficiency and strong practicability.
Description
Technical Field
The invention relates to the technical field of water treatment, in particular to a biological algicide and a preparation method and application thereof.
Background
With the increasing progress of urban mass, the water is enriched with a great amount of nutrient elements (especially nitrogen and phosphorus), which continuously promotes the accelerated development of primary production and algae eutrophication. The water eutrophication is mainly characterized in that a plurality of phytoplankton is subjected to explosive increment in a large range and short time under the condition of sufficient nutrition, and even is gathered in a high density, so that the transparency of water quality is reduced, and the disastrous water ecology abnormal phenomenon of the normal life activities of aquatic organisms is caused. Eutrophication is favorable for periodic propagation and dominant propagation environments of plankton such as blue algae and benthos, and the like, and the microcystis aeruginosa bloom is often exploded in China, so that the problem of large-scale water resource pollution is caused.
The algae removal methods adopted at home and abroad at present comprise a physical method, a chemical method and a biological method. The physical method mainly comprises mechanical dredging, algae filtering, clay algae removal, air floatation algae removal and the like, has high cost and high cost, is difficult to apply to blue algae control with a large area, and is limited in practical application; the chemical method mainly adopts chemical agents such as copper sulfate, hydrogen peroxide, surfactant, sodium hypochlorite and the like, and the method is easy to produce secondary pollution and generates potential harm to water organisms. The biological method is to control the increase of algae number by utilizing natural enemies of algae such as bacteria, viruses, zooplankton preying on red tide organisms, aquatic plants, macroalgae and the like or certain substances generated by the algae for inhibiting the growth of the algae so as to achieve the aim of treatment.
Disclosure of Invention
In order to overcome the defects and shortcomings in the prior art, the invention aims to provide the biological algicide, which is obtained by modifying chitosan after fermenting and culturing the composite microbial inoculum, and has strong environment change resistance and high algicide efficiency in the actual use process.
The invention also aims to provide a preparation method of the biological algicide, which comprises the steps of fermenting and culturing the composite microbial inoculum, and adopting chitosan modified composite microbial inoculum to prepare the biological algicide; the preparation method has the advantages of simple operation, low cost and no secondary pollution, and the prepared biological algicide has high algicide efficiency.
The aim of the invention is achieved by the following technical scheme: a biological algicide is obtained by modifying chitosan after fermenting and culturing a composite microbial inoculum, wherein the composite microbial inoculum comprises actinomycetes, bacillus, alteromonas and myxobacteria.
The microbial inoculum of the biological algicide adopted by the invention is a composite microbial inoculum, and the prepared composite microbial inoculum has stronger environmental change tolerance capability and higher algicide treatment efficiency in the actual use process by matching actinomycetes, bacillus, alteromonas and myxobacteria.
Further, the composite microbial inoculum comprises the following components in parts by weight: 3 to 5 parts of actinomycetes, 3 to 5 parts of bacillus, 1 to 3 parts of alteromonas and 0.5 to 1 part of myxobacteria.
Further, the composite microbial inoculum comprises the following components in parts by weight: 3 to 5 parts of streptomyces microflavus, 3 to 5 parts of bacillus subtilis, 1 to 3 parts of pseudoalteromonas hainana and 0.5 to 1 part of myxococcus aurantiacus.
Further, actinomycetes are Streptomyces microflavus (Streptomyces microflavus) (ACCC 40027); the bacillus adopts bacillus subtilis (Bacillus subtilis) (CGMCC 1.9083); alteromonas is pseudoalteromonas maritima (Pseudoalteromonas marina) (CGMCC 1.15847); myxobacteria were myxococcus aurantiacus (Myxococcus xanthus, CGMCC 1.3865).
According to the invention, the microbial inoculum is matched and the dosage proportion is controlled, so that each microbial inoculum cooperatively grows and cooperatively treats algae in an algae outbreak water body, and the prepared composite microbial inoculum has strong environmental change resistance in the actual use process and has excellent algae removal effect.
The other object of the invention is achieved by the following technical scheme: the preparation method of the biological algicide comprises the following steps:
(1) Fermenting and culturing the prepared composite microbial inoculum to obtain composite bacterial liquid;
(2) Mixing the compound bacterial liquid in the step (1) with the chitosan solution in proportion, and adopting the chitosan modified compound bacterial agent to obtain the biological algicide.
In the preparation method of the biological algicide, the composite microbial inoculant is fermented and cultured, and then the chitosan modified composite microbial inoculant is adopted to prepare the biological algicide, so that the algicide effect of the composite microbial inoculant can be further improved. The preparation method is simple and convenient to operate, low in production cost and high in production efficiency, and the prepared biological algicide is high in algicide efficiency and high in practicability.
Further, in the step (1), the fermentation culture method of the composite microbial inoculum comprises the following steps:
a1, inoculating the composite microbial inoculum into a shake flask containing a culture medium, shake-culturing in an incubator at 20-40 ℃ for 24-48 hours to obtain primary seed liquid, and controlling the rotating speed at 100-200 rpm;
a2, willInoculating the primary seed liquid obtained in the step A1 into a fermentation tank according to the inoculum size with the volume fraction of 5-20%, and performing semi-continuous culture fermentation under the culture conditions of 20-40 ℃ and the rotation speed of 100-200 rpm, pH of 6-9 and dissolved oxygen of 10-50%; after fermentation, the solid content of the thalli is 50-80%, and the microbial mass is 1.0 x 10 11 ～5.0*10 11 cfu/mL。
The biological algicide achieves the algae inhibiting effect mainly by secreting extracellular active substances, and the secretion amount of the extracellular active substances is closely related to the microbial biomass; the invention creatively adopts semi-continuous fermentation, can realize high-density fermentation of microorganisms, obtains more fermentation products and ensures removal efficiency.
Further, in the step A1, the medium includes the following raw materials in concentration: 1 to 5g/L of glucose, 2 to 20g/L of starch, 0.05 to 0.5g/L of magnesium sulfate heptahydrate, 0.05 to 2g/L of monopotassium phosphate, 0.05 to 2g/L of monoammonium phosphate, 0.05 to 2g/L of corn steep liquor dry powder, 5 to 20g/L of sodium chloride and 0.05 to 2 percent of microelement agent. The addition amount of the trace element agent is 0.05-2% by volume.
Further, the microelement agent comprises the following raw materials in concentration: ferrous sulfate heptahydrate 0.1-0.5 g/L, manganese chloride 0.5-3 g/L, anhydrous copper sulfate 0.02-0.1 g/L, sodium molybdate 0.2-0.5 g/L, and zinc sulfate 0.05-0.2 g/L.
Further, in the step A2, the semi-continuous fermentation step is to discharge 20-80% of fermentation liquor by volume percentage every 8-36 h, supplement the culture medium with the same volume, control the fermentation system by pH in the fermentation process, supplement the carbon source when the pH is reduced, the pH is increased, supplement the nitrogen source, take 3-5 times, end the fermentation, and supplement the culture medium by volume percentage after the fermentation is ended by 5-20%. By adopting the steps to perform semi-continuous fermentation, the invention can realize high-density fermentation of microorganisms and obtain more fermentation products, so that the prepared biological algicide has high algae removal efficiency.
Further, in the step (2), the method for modifying the composite microbial inoculum by using chitosan comprises the following steps:
b1, adding chitosan into deionized water, and dropwise adding glacial acetic acid to prepare a chitosan solution;
and B2, mixing the composite bacterial solution and the chitosan solution in proportion to obtain the biological algicide.
In step B2, the composite bacterial solution and the chitosan solution are stirred and mixed according to the volume ratio of 0.25-4:1, and are mixed at the temperature of (-2) to (-6) DEG C, so as to obtain the biological algicide.
The chitosan adopted by the invention is a natural high molecular organic compound, is nontoxic and harmless, can be biodegraded in the environment, contains a large amount of free amino acids in chitosan molecules, and can be hydrolyzed into NH3 < + > in a dilute acid solution, so that the chitosan has a large amount of positive charges, the chitosan shows the performance of cationic polyelectrolyte, and the chitosan can be electrostatically adsorbed with algae cells with negative charges to form floccules, thereby achieving the aim of removing algae. According to the invention, the chitosan modified composite microbial agent is adopted, so that the algae removal effect of the composite microbial agent can be further improved.
The invention also provides an application of the biological algaecide, the obtained biological algaecide is put into an algae burst water body, and the adding amount of the biological algaecide in the water body is as follows: in the initial stage of algae burst, the bacterial dosage is 1-5 mL/m 3 A body of water; in the middle period of algae burst, the bacterial dosage is 10-100 mL/m 3 A body of water; in the late stage of algae burst, the bacterial dosage is 5-50 mL/m 3 And (3) a water body. When the biological algicide is applied to algae outbreak water, the biological algicide has the advantages of simplicity in operation, low cost, no secondary pollution and high algae removal efficiency.
The invention has the beneficial effects that: according to the invention, after the composite microbial inoculum is fermented and cultured, the chitosan modified composite microbial inoculum is adopted to prepare the biological algicide, the microbial inoculum of the biological algicide is the composite microbial inoculum, and the composite microbial inoculum has stronger environmental change resistance and higher treatment efficiency in the actual use process; the biological algicide achieves the algae inhibition effect mainly by secreting extracellular active substances, the secretion amount of the extracellular active substances is closely related to the microbial biomass, the semi-continuous fermentation creatively adopted by the invention can realize the high-density fermentation of microorganisms, obtain more fermentation products and ensure the removal efficiency; the chitosan modified composite microbial inoculum is adopted, so that the algae removal effect of the composite microbial inoculum can be further improved.
Detailed Description
The invention will be further illustrated by the following examples, which are not intended to limit the scope of the invention, in order to facilitate the understanding of those skilled in the art.
In a typical embodiment of the invention, a biological algaecide is obtained by fermenting and culturing a composite microbial agent, wherein the composite microbial agent comprises actinomycetes, bacillus, alteromonas and myxobacteria, and then modifying chitosan.
Further, the composite microbial inoculum comprises the following components in parts by weight: 3 to 5 parts of actinomycetes, 3 to 5 parts of bacillus, 1 to 3 parts of alteromonas and 0.5 to 1 part of myxobacteria.
Further, the composite microbial inoculum comprises the following components in parts by weight: 3 to 5 parts of streptomyces microflavus, 3 to 5 parts of bacillus subtilis, 1 to 3 parts of pseudoalteromonas hainana and 0.5 to 1 part of myxococcus aurantiacus. In the embodiment of the invention, actinomycetes: streptomyces microflavus (Streptomyces microflavus) (accession number ACCC 40027); bacillus: bacillus subtilis (Bacillus subtilis) (CGMCC 1.9083); alteromonas: pseudomonas seai (Pseudoalteromonas marina) (CGMCC 1.15847); myxobacteria: myxococcus orange (Myxococcus xanthus, CGMCC 1.3865).
In an exemplary embodiment of the present invention, a method for preparing the above biological algicide includes the following steps:
(1) Fermenting and culturing the prepared composite microbial inoculum to obtain composite bacterial liquid;
(2) Mixing the compound bacterial liquid in the step (1) with the chitosan solution in proportion, and adopting the chitosan modified compound bacterial agent to obtain the biological algicide.
Further, in the step (1), the fermentation culture method of the composite microbial inoculum comprises the following steps:
a1, inoculating the composite microbial inoculum into a shake flask containing a culture medium, shake-culturing in an incubator at 20-40 ℃ for 24-48 hours to obtain primary seed liquid, and controlling the rotating speed at 100-200 rpm;
a2, inoculating the primary seed liquid obtained in the step A1 into a fermentation tank according to the inoculum size with the volume fraction of 5-20%, and performing semi-continuous culture fermentation under the culture conditions of 20-40 ℃ and the rotation speed of 100-200 rpm, pH of 6-9 and dissolved oxygen of 10-50%; after fermentation, the solid content of the thalli is 50-80%, and the microbial mass is 1.0 x 10 11 ～5.0*10 11 cfu/mL。
Further, in the step A1, the medium includes the following raw materials in concentration: 1 to 5g/L of glucose, 2 to 20g/L of starch, 0.05 to 0.5g/L of magnesium sulfate heptahydrate, 0.05 to 2g/L of monopotassium phosphate, 0.05 to 2g/L of monoammonium phosphate, 0.05 to 2g/L of corn steep liquor dry powder, 5 to 20g/L of sodium chloride and 0.05 to 2 percent of microelement agent. The addition amount of the trace element agent is 0.05-2% by volume.
Further, the microelement agent comprises the following raw materials in concentration: ferrous sulfate heptahydrate 0.1-0.5 g/L, manganese chloride 0.5-3 g/L, anhydrous copper sulfate 0.02-0.1 g/L, sodium molybdate 0.2-0.5 g/L, and zinc sulfate 0.05-0.2 g/L.
Further, in the step A2, the semi-continuous fermentation step is to discharge 20-80% of fermentation liquor by volume percentage every 8-36 h, supplement the culture medium with the same volume, control the fermentation system by pH in the fermentation process, supplement the carbon source when the pH is reduced, the pH is increased, supplement the nitrogen source, take 3-5 times, end the fermentation, and supplement the culture medium by volume percentage after the fermentation is ended by 5-20%.
Further, in the step (2), the method for modifying the composite microbial inoculum by using chitosan comprises the following steps:
b1, adding chitosan into deionized water, and dropwise adding glacial acetic acid to prepare a chitosan solution;
and B2, mixing the composite bacterial solution and the chitosan solution in proportion to obtain the biological algicide.
In step B2, the composite bacterial solution and the chitosan solution are stirred and mixed according to the volume ratio of 0.25-4:1, and are mixed for 22-26 hours at the temperature of (-2) to (-6) DEG C, so as to obtain the biological algicide.
The invention also provides an application of the biological algaecide, the obtained biological algaecide is put into an algae burst water body, and the adding amount of the biological algaecide in the water body is as follows: in the initial stage of algae burst, the bacterial dosage is 1-5 mL/m 3 A body of water; in the middle period of algae burst, the bacterial dosage is 10-100 mL/m 3 A body of water; in the late stage of algae burst, the bacterial dosage is 5-50 mL/m 3 And (3) a water body.
Example 1
In this embodiment, the fermentation culture method of the composite microbial inoculum comprises the following steps:
a1, inoculating a composite microbial inoculum comprising actinomycetes, bacillus, alteromonas and myxobacteria into a shake flask containing a culture medium, wherein the inoculation amount is 10%, shake-culturing in an incubator at 30 ℃ for 24 hours to obtain a first-stage seed liquid, and controlling the rotating speed to 150rpm; the composite microbial inoculum comprises the following components in parts by weight: 4 parts of actinomycetes, 4 parts of bacillus, 2 parts of alteromonas and 0.8 part of myxobacteria.
A2, inoculating the seed liquid obtained in the step A1 into a fermentation tank according to the inoculation amount with the volume fraction of 10%, and performing semi-continuous culture fermentation under the culture conditions of 30 ℃ and 150rpm of rotation speed, 6.8 pH and 30% of dissolved oxygen.
Further, the semi-continuous fermentation step comprises the steps of discharging 20% of fermentation liquor by volume percentage every 8h, supplementing the culture medium with the same volume, controlling a fermentation system by pH in the fermentation process, supplementing a carbon source when the pH is reduced, supplementing a pH rising, supplementing a nitrogen source, carrying out belt discharge for 5 times, ending the fermentation, supplementing 10% of the culture medium by volume percentage after ending the fermentation, obtaining a compound microbial inoculum with the solid content of 70% and the microbial biomass of 3.2 x 10 11 cfu/mL。
Further, the culture medium of the composite microbial inoculum is as follows: 2g/L of glucose, 5g/L of starch, 0.1g/L of magnesium sulfate heptahydrate, 0.1g/L of monopotassium phosphate, 0.2g/L of monoammonium phosphate, 0.08g/L of corn steep liquor dry powder, 10g/L of sodium chloride and 0.5% of trace element agent.
Further, the microelement agent comprises the following raw materials with the following concentration: ferrous sulfate heptahydrate 0.2g/L, manganese chloride 1g/L, anhydrous copper sulfate 0.05g/L, sodium molybdate 0.2g/L, and zinc sulfate 0.1g/L.
Example 2
In this embodiment, the method for modifying the composite microbial inoculum by using chitosan comprises the following steps:
b1, preparation of chitosan solution
100mg of chitosan is weighed, 20mL of distilled water is added, a small amount of glacial acetic acid is added dropwise to dissolve the chitosan, and finally the volume is complemented to 100mL, so that 1mg/mL of chitosan solution is obtained.
B2 and chitosan modified composite microbial agent
The composite bacterial solution and chitosan solution obtained in the example 1 are mixed according to the volume ratio of 4:1, and mixing for 24 hours at the temperature of-4 ℃ to obtain the modified composite microbial inoculum, namely the obtained biological algicide.
Example 3
This embodiment differs from embodiment 1 in that: in this embodiment, the fermentation culture method of the composite microbial inoculum comprises the following steps:
a1, inoculating a composite microbial inoculum comprising actinomycetes, bacillus, alteromonas and myxobacteria into a shake flask containing a culture medium, wherein the inoculation amount is 12%, shake-culturing in an incubator at 25 ℃ for 36 hours to obtain a first-stage seed liquid, and controlling the rotating speed at 100rpm; the composite microbial inoculum comprises the following components in parts by weight: 3 parts of actinomycetes, 3 parts of bacillus, 1 part of alteromonas and 0.5 part of myxobacteria.
A2, inoculating the seed solution obtained in the step A1 into a fermentation tank according to the inoculation amount with the volume fraction of 15%, and performing semi-continuous culture fermentation under the culture conditions of 25 ℃ and the rotation speed of 100rpm, pH7.2 and 20% of dissolved oxygen.
Further, the semi-continuous fermentation step is to discharge 30% of fermentation liquor by volume percentage every 10h, supplement the culture medium by the same volume, control the fermentation system by pH in the fermentation process, supplement the carbon source when the pH is reduced, supplement the pH is increased, supplement the nitrogen source, carry out 4 times of belt discharge, finish the fermentation, and supplement the culture medium by volume percentage by 15% after finishing the fermentation.
The rest of this embodiment is the same as embodiment 1, and will not be described here again.
Example 4
This embodiment differs from embodiment 1 in that: in this embodiment, the fermentation culture method of the composite microbial inoculum comprises the following steps:
a1, inoculating a composite microbial inoculum comprising actinomycetes, bacillus, alteromonas and myxobacteria into a shake flask containing a culture medium, wherein the inoculation amount is 15%, shake-culturing in an incubator at 20 ℃ for 42 hours to obtain a first-stage seed liquid, and controlling the rotating speed at 200rpm; the composite microbial inoculum comprises the following components in parts by weight: 5 parts of actinomycetes, 5 parts of bacillus, 3 parts of alteromonas and 1 part of myxobacteria.
A2, inoculating the seed solution obtained in the step A1 into a fermentation tank according to the inoculation amount with the volume fraction of 20%, and performing semi-continuous culture fermentation under the culture conditions of the fermentation tank that the temperature is 35 ℃, the rotating speed is 200rpm, the pH value is 7.6 and the dissolved oxygen is 24%.
Further, the semi-continuous fermentation step is to discharge 25% of fermentation liquor by volume percentage every 16h, supplement the culture medium by the same volume, control the fermentation system by pH in the fermentation process, supplement the carbon source when the pH is reduced, supplement the pH is increased, supplement the nitrogen source, carry out 4 times of belt discharge, finish the fermentation, and supplement 20% of the culture medium by volume percentage after finishing the fermentation.
The rest of this embodiment is the same as embodiment 1, and will not be described here again.
Example 5
This embodiment differs from embodiment 2 in that: the volume ratio of the composite bacterial liquid obtained in the example 3 to the chitosan solution is 0.25:1, mixing at the temperature of minus 6 ℃ for 26 hours to obtain the modified composite microbial inoculum, namely the obtained biological algicide.
The rest of this embodiment is the same as embodiment 2, and will not be described here again.
Example 6
This embodiment differs from embodiment 2 in that: the composite bacterial solution and chitosan solution obtained in the example 4 are mixed according to the volume ratio of 2:1, and mixing at the temperature of minus 2 ℃ for 22 hours to obtain the modified composite microbial inoculum, namely the obtained biological algicide.
The rest of this embodiment is the same as embodiment 2, and will not be described here again.
Example 7
The biological algaecide obtained in the embodiment 2 is put into a water body of algae burst, and the removing effect is as follows:
TABLE 1 algae removal rate of biological algicides at the early stage of algae eruption%
TABLE 2 algae removal rate of biological algaecides in the middle of algae burst%
TABLE 3 algae removal rate of biological algicides at the late stage of algae eruption%
Wherein the initial stage of algae burst in the table is that the adding amount of the biological algicide, namely the bacterial dosage is 3mL/m 3 A body of water; in the middle period of algae burst, the bacterial dosage is 50mL/m 3 A body of water; in the late stage of algae burst, the bacterial dosage is 20mL/m 3 And (3) a water body.
The biological algicide is obtained by modifying chitosan after fermenting and culturing the composite microbial inoculum, has strong environmental change tolerance in the actual use process, has the advantages of simple operation, low cost and no secondary pollution when being applied to algae removal in an algae outbreak water body, and has high algae removal efficiency and strong practicability.
The above embodiments are preferred embodiments of the present invention, and besides, the present invention may be implemented in other ways, and any obvious substitution is within the scope of the present invention without departing from the concept of the present invention.
Claims (7)
1. A preparation method of a biological algicide is characterized by comprising the following steps: the method comprises the following steps:
(1) Fermenting and culturing the composite microbial inoculum to obtain composite bacterial liquid; after fermentation, the solid content of the thalli is 50-80%, and the microbial mass is 1.0 x 10 11 ～5.0*10 11 cfu/mL; the composite microbial inoculum comprises the following components in parts by weight: 3 to 5 parts of actinomycetes, 3 to 5 parts of bacillus, 1 to 3 parts of alteromonas and 0.5 to 1 part of myxobacteria;
(2) Mixing the compound bacterial liquid in the step (1) with the chitosan solution in proportion, and adopting the chitosan modified compound bacterial agent to obtain the biological algicide;
in the step (2), the method for modifying the composite microbial inoculum by using the chitosan comprises the following steps:
b1, adding chitosan into deionized water, and dropwise adding glacial acetic acid to prepare a chitosan solution;
and B2, mixing the composite bacterial solution and the chitosan solution in proportion to obtain the biological algicide.
2. The method for preparing the biological algicide according to claim 1, wherein: in the step (1), the fermentation culture method of the composite microbial inoculum comprises the following steps:
a1, inoculating the composite microbial inoculum into a shake flask containing a culture medium, shake-culturing in an incubator at 20-40 ℃ for 24-48 hours to obtain primary seed liquid, and controlling the rotating speed at 100-200 rpm;
a2, inoculating the primary seed liquid obtained in the step A1 into a fermentation tank according to the inoculum size with the volume fraction of 5-20%, and performing semi-continuous culture fermentation under the culture conditions of the fermentation tank, wherein the temperature is 20-40 ℃, the rotating speed is 100-200 rpm, the pH value is 6-9 and the dissolved oxygen is 10-50%.
3. The method for preparing the biological algicide according to claim 2, wherein: in the step A1, the culture medium comprises the following raw materials in concentration: 1 to 5g/L of glucose, 2 to 20g/L of starch, 0.05 to 0.5g/L of magnesium sulfate heptahydrate, 0.05 to 2g/L of monopotassium phosphate, 0.05 to 2g/L of monoammonium phosphate, 0.05 to 2g/L of corn steep liquor dry powder, 5 to 20g/L of sodium chloride and 0.05 to 2 percent of microelement agent.
4. The method for preparing the biological algicide according to claim 2, wherein: in the step A2, the semi-continuous fermentation step is to discharge 20-80% of fermentation liquor by volume percentage every 8-36 h, supplement the culture medium with the same volume, control the fermentation system by pH in the fermentation process, supplement the carbon source when the pH is reduced, supplement the pH is increased, supplement the nitrogen source, take 3-5 times, finish the fermentation, and supplement the culture medium by volume percentage by 5-20% after finishing the fermentation.
5. The method for preparing the biological algicide according to claim 1, wherein: in the step B2, the composite bacterial solution and the chitosan solution are stirred and mixed according to the volume ratio of 0.25-4:1, and are mixed at the temperature of (-2) to (-6) DEG C, so as to obtain the biological algicide.
6. A biological algaecide, which is characterized in that: the biological algaecide is prepared by the preparation method of the biological algaecide according to any one of claims 1 to 5.
7. Use of the biological algaecide according to claim 6, wherein: the biological algicide is put into an algae burst water body, and the adding amount of the biological algicide in the water body is as follows: in the initial stage of algae burst, the bacterial dosage is 1-5 mL/m 3 A body of water; in the middle period of algae burst, the bacterial dosage is 10-100 mL/m 3 A body of water; in the late stage of algae burst, the bacterial dosage is 5-50 mL/m 3 And (3) a water body.
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| CN111849826A (en) * | 2020-07-30 | 2020-10-30 | 合肥市东方美捷分子材料技术有限公司 | Composite microbial agent for blue algae inhibition and preparation method and application thereof |
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