CN107986456A - A kind of prevention and suppressing method applied to water body cyanobacteria - Google Patents

A kind of prevention and suppressing method applied to water body cyanobacteria Download PDF

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CN107986456A
CN107986456A CN201711334524.4A CN201711334524A CN107986456A CN 107986456 A CN107986456 A CN 107986456A CN 201711334524 A CN201711334524 A CN 201711334524A CN 107986456 A CN107986456 A CN 107986456A
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prevention
waters
chlorella
cyanobacteria
bioactivity
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徐孟
金波
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Taizhou Yongda Biotechnology Co Ltd
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Taizhou Yongda Biotechnology Co Ltd
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    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • 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
    • C02F3/32—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae
    • C02F3/322—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae use of algae
    • C02F3/325—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae use of algae as symbiotic combination of algae and bacteria
    • 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
    • C02F3/347—Use of yeasts or fungi
    • 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
    • C02F3/348—Biological treatment of water, waste water, or sewage characterised by the microorganisms used characterised by the way or the form in which the microorganisms are added or dosed
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2303/00—Specific treatment goals
    • C02F2303/20—Prevention of biofouling

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  • Life Sciences & Earth Sciences (AREA)
  • Microbiology (AREA)
  • Engineering & Computer Science (AREA)
  • Water Supply & Treatment (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Hydrology & Water Resources (AREA)
  • Environmental & Geological Engineering (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • Botany (AREA)
  • Ecology (AREA)
  • Mycology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention belongs to cyanobacteria prevention and processing applied technical field, specifically disclose a kind of prevention applied to water body cyanobacteria and suppressing method, step 1, determine to apply the waters of bioactivity bacteria agent product first, recycles sprinkling machinery that bioactivity bacteria agent is uniformly applied to pending waters.Step 2, be monitored pending waters, 78 it is small when after, then pending waters pass through spray machinery apply chlorella solution.Step 3, be monitored the waters for spraying chlorella solution, according to the cyanobacteria state of detection, repeat step 1, step 2.The beneficial effects of the present invention are：Pass through the bioactivity bacteria agent of high-content, energy fast decoupled applies the organic matter in waters, application is supplemented at the same time beneficial to chlorella solution, beneficial bacterium is set quickly to absorb the nutriment in applied waters, so as to quickly breed, harmful algae living space is reduced high-content bacillus and decompose organic matter, nitrogen in waters, phosphorus ratio are applied in adjusting, realize prevention and the purpose administered.

Description

A kind of prevention and suppressing method applied to water body cyanobacteria
Technical field
The invention belongs to cyanobacteria prevention and processing applied technical field, and in particular to a kind of prevention applied to water body cyanobacteria And suppressing method.
Background technology
Cyanobacteria is prokaryotes, is called blue-green alge or cyanobacteria；There is colloid clothing outside the cell membrane of most of cyanobacterias, be called Myxophyceae.In algae, cyanobacteria is most simple, the unicellular organism of most original, such as Microcystis aeruginosa, nostoc, anabena.Cyanobacteria does not have There is nucleus, there is nuclear matter in cell center, is usually in granular form or netted, and pigment is evenly distributed in cytoplasm, nuclear matter There is no nuclear membrane and kernel, there is core, therefore referred to as protokaryon（Or nucleoid）, have cyclic DNA plasmid in cyanobacteria, take on carrier Effect.
In some water bodys full of nutrition, some cyanobacterias often in summer amount reproduction, and the water surface formed one layer it is bluish-green Color and the offscum for having bad smell, are known as " wawter bloom ", large-scale blue algae bloom, is referred to as " green tide "（The red tide occurred with ocean It is corresponding）.Green tide causes water quality deterioration, exhausts water oxygen when serious and causes the death of fish, more seriously, in cyanobacteria Some species（Such as Microcystis aeruginosa）Microcystin can also be produced（Microcystins, abbreviation MCs）, contain in about 50% green tide There are a large amount of MCs.MCs to fish, people and animals in addition to directly producing murder by poisoning, and the major incentive of liver cancer.MCs is heat-resisting, is not easy Decomposed by boiling water, but can be by active carbon absorption, it is possible to contaminated water source is purified with activated carbon water purifier.
When cyanobacteria largely occurs, neighbouring water body is generally covered in blueness or green, the water surface by thick blue-green water bloom, Accumulated by wind to bank, can not only give out a foul smell taste, and the Cells of Blue-green Algae containing toxin floats in water body, when with some suspensions Thing complex-precipitation, or precipitated after being preyed on by cultivation object with its excreta, in fishpond, bottom of pond is enriched with, and nuisanceless aquatic products are produced Huge negative effect can be brought.
Anabaena in cyanobacteria can quickly produce lethal factor, destroy the gill tissue of cultivation object, disturb its metabolism Be normally carried out, paralysis nerve, makes its dead, indivedual kinds not only live body bands poison, and dead individuals are decomposed and can produced in cyanobacteria Biotoxin-cyanophycean toxin（Such as Microcystin）.Cyanophycean toxin amount can directly contribute cultivation object and be poisoned to death when more；Or Even if person's quantity is few, cultivation object can be also endangered by food chain build-up effect, until harmful to human.
Therefore, a kind of prevention and suppressing method applied to water body cyanobacteria is provided based on the above problem, the present invention.
The content of the invention
Goal of the invention：, can be to not the object of the present invention is to provide a kind of prevention applied to water body cyanobacteria and suppressing method The waters that cyanobacteria occurs is prevented, the waters that cyanobacteria has occurred is carried out efficiently, the processing stablized and without any negative effect, Ensure the safety of water body environment.
Technical solution：The present invention provides a kind of prevention and suppressing method applied to water body cyanobacteria, comprises the following steps, and walks Rapid 1, the waters of application bioactivity bacteria agent product is determined first, recycles sprinkling machinery uniformly to apply bioactivity bacteria agent Pending waters is added in, wherein, bioactivity bacteria agent is with bacillus subtilis（Bacillus subtilis）, Amur it is false Monad（Pseudomonas stutzeri）, candida aquatica（Candida aquatica）For strain, use is organic, nothing Machine raw material, is formulated compound after each strain individually fermentation.Step 2, be monitored pending waters, when 7-8 is small after, then Apply chlorella solution by spraying machinery in pending waters, wherein, the chlorella content of chlorella solution is 3~8 × 107 Cfu/ml, pH value are 6.5-9.5.Step 3, to spray chlorella solution waters be monitored, when 6-7 is small after, according to monitoring The cyanobacteria state of detection, repeat step 1, the processing step of step 2.
The technical program, the bioactivity bacteria agent in the step 1, effective total viable count（Containing bacillus subtilis, Pseudomonas stutzeri, candida aquatica）≥2×108 A/mL, bacillus number >=1 × 108A/mL, wherein, it is effectively living Bacterium is containing bacillus subtilis, Pseudomonas stutzeri, candida aquatica.
The technical program, in the step 3, before low temperature drying, uniformly sprayed on the cereal for treat low temperature drying vaporific Water, sprinkling water temperature is 10 degrees Celsius -12 degrees Celsius, then natural air drying -60 minutes 30 minutes in warehouse.
The technical program, the pH of bioactivity bacteria agent is 4-6.5 in the step 1, the sum of mould miscellaneous bacteria for≤ 3.0×105 Cfu/ml, coliform are≤10 MPN/100mL；In the step 1 bioactivity bacteria agent spray when, it is necessary to The living bacteria count of microbial inoculum is measured, is comprised the following steps, the sample 25mL fully mixed, be put into sterile working by a 1 is made in sterilizing conical flask containing 225mL sterile biological brine：10 uniform dilution.B, 1 is drawn with 1mL sterilized straws： 10 dilution 1mL, inject in the test tube containing 9mL sterile salines, shaking mixes, and makes 1 slowly along tube wall：100 Dilution.C, 1mL sterilized straws separately are taken, by aforesaid operations order, does 10 times of incremental dilutions, be so often incremented by once, that is, change With 1 1mL sterilized straw.D, 1 is selected：10-7、1：10-8Two acceptable diluent degree, respectively take 1mL to be separately added into counting culture Base plate, each dilution factor make three plates.E, dilution is moved into plate, in time should be noted the counting culture medium for being cooled to 45 DEG C Enter plate about 15mL-20mL, and rotate plate to make to be uniformly mixed, while be poured into culture medium is counted added with 1mL dilution samples Make blank control in the sterilizing plates of sterile saline, bacillus subtilis, Pseudomonas stutzeri are trained using glucose Base culture is supported, candida aquatica uses starch culture-medium culture.F, after agar completely solidification after, overturn tablet, put 32 DEG C ± Culture 48h ± 2h in 1 DEG C of incubator, chooses tablet of the clump count between 30-300 and is counted.G, bacterium number calculates, according to every Then one dilution factor is multiplied by the extension rate of sample, is obtained viable count in every milliliter of sample using the average of three flat-plate bacterial colonies； If any two dilution factors, how the bacterium colony of its growth determines between 30-300 depending on ratio between two, as its ratio is less than 2, It should report its average；Such as larger than 2, then report wherein less numeral.
The technical program, when bioactivity bacteria agent sprays in the step 1, it is necessary to the living bacteria count of microbial inoculum into Row measure, comprises the following steps, and a, will first pour into 10 mm cuvettes for cultivating the fluid nutrient medium of strain, selects 600nm Zero point is adjusted on wavelength spectrophotometer, as blank control.B, 300 mL of sample is put into sterilizing wide-mouth bottle again, up and down acutely Shaking 30 times -40 times, is suctioned out in injection 10mm cuvettes with sterilized straw, measures the OD value under 600nm wavelength.
The technical program, the component of starch culture-medium is starch 20g, potassium nitrate 1.0g, phosphoric acid hydrogen two in the step 1 Potassium 0.5g, sodium chloride 0.5g, magnesium sulfate 0.5g, ferric sulfate 0.01g, after mixing plus distilled water is first to 1000mL, pH7.2 ± 0.1 First all of above component is added in distilled water, by 2% addition agar, dissolving is reheated, corrects pH 7.2 ± 0.1, finally divide Dress, 121 DEG C, time 15min-20min of autoclaving, heating fusing agar, uses when being cooled to 45 DEG C during use.
The technical program, in the step 1 glucose cultivate the glucose 6g of base, ammonium chloride 1.0g, sodium chloride 1.0g, 1.0 g of dipotassium hydrogen phosphate, potassium dihydrogen phosphate 1.0g, magnesium sulfate 0.2g, after mixing plus distilled water is to 1000mL, pH7.2 ± 0.1, All of above component is added in distilled water first, by 2% addition agar, dissolving is reheated, corrects pH 7.2 ± 0.1, finally Packing, 121 DEG C of autoclaving, time be 15min-20 min, during use heating melt agar, used when being cooled to 45 DEG C.
The technical program, it is necessary to be carried out to the mould miscellaneous bacteria of microbial inoculum when bioactivity bacteria agent sprays in the step 1 Measure, is tested, composition is potassium dihydrogen phosphate 1.0g, glucose 10.0g, first adds all of above composition using Martin's culture medium Enter in distilled water, then add rose-bengal aqueous solution, separately dissolve chloramphenicol with a small amount of ethanol, add in culture medium and dissolve by heating, most Packing, 121 DEG C of autoclaving, time be 15min-20 min afterwards, during use heating melt agar, used when being cooled to 45 DEG C.
The technical program, it is necessary to chlorella content in chlorella solution when spraying chlorella solution in the step 2 Tested, comprised the following steps, a, microscopy counting chamber, before sample-adding, first carry out microscopy, observation is counted to the counting chamber of tally Answer no-sundries in room.B, be loaded, dry blood counting chamber covered will be cleaned, with sterile pipette by bead algae solution by Coverslip edge drips a droplet, allows algae solution to be advanced into counting chamber certainly by capillary osmosis along gap, can not there is bubble generation.c、 Microscopic counting, after standing 5min, blood counting chamber is placed on microscope carrier, first finds counting chamber with low power lens, It is converted into high power lens to be counted, counting chamber is selected 5 middle lattice and counted.D, calculate, count the number of chlorella in lattice in 5 Measure A, then in algae solution chlorella quantity=A/80 × 100 × 10000 × extension rate/mL, wherein, blood counting chamber for 25 × 16, counting chamber thickness is 0.1mm.E, clear up, it is used during blood counting chamber, beaker, test tube, pipette, microscope etc. are tested To article clean out.F, the measure of pH values, takes and pours into right amount in beaker, and pH is directly measured with the acidometer of calibrated mistake Value.
The technical program, it is necessary to carry out fresh water culture when the chlorella content in the chlorella solution is tested Base, wherein, cultivate reagent component is NaNO3, K2HPO4, MgSO47H2O, CaCl22H2O, citric acid, ferric citrate Or ironic citrate, Na2EDTA, Na2CO3, A5, wherein, it is each form content be respectively 1.5000g/L, 0.0400g/L, 0.0750g/L, 0.0360 g/L, 0.0060g/L, 0.006g/L or 0.004g/L, 0.0010g/L, 0.0200g/L, 1.0mL； The A5 by ZnSO47H2O, CuSO45H2O, MoO3, H3BO3, MnCl24H2O and content be respectively 0.222g/L, 0.079g/L、0.015g/L、2.86g/L、1.81g/L。
The technical program, it is necessary to carry out cultivation in sea water when the chlorella content in the chlorella solution is tested Base, wherein, be made of liquid A and liquid B, solution A agent formulations for FeCl36H2O, MnCl24H2O, H3BO3, Na2EDTA, NaH2PO42H2O, NaNO3 and content are respectively 6.5g, 1.8g, 168g, 225g, 100g, 500g, and second liquid agent formulations are ZnCl2, CoCl26H2O, (NH4) 6Mo7O244H2O, CuSO45H2O and content be respectively 2.1g, 2.0g, 0.9g, 2.0g, preparation method is first to be dissolved in solution A in 5L pure water by content sequence, then second liquid is dissolved in 100ml by content sequence In pure water, then solution A and second liquid sterilized, then by the liquid A and liquid B after sterilizing, solution A：Second liquid=1000:1 or 500：1.
Compared with prior art, it is of the invention a kind of to exist applied to the prevention of water body cyanobacteria and the beneficial effect of suppressing method In：Pass through the bioactivity bacteria agent of high-content（Bacillus content 10,000,000,000）, energy fast decoupled is applied organic in waters Matter, while application is supplemented beneficial to chlorella solution, beneficial bacterium is quickly absorbed the nutriment in applied waters, so that quick numerous Grow, harmful algae living space is reduced high-content bacillus and decompose organic matter, be also adjusted in synchronism and apply nitrogen in waters, phosphorus ratio Example, it is final so as to apply N∶P ratio in waters normal, so that effectively decrease cyanobacteria produces by promoting nitrogen with phosphorus, with nitrogen consolidate phosphorus Condition, realize efficiently prevent and administer purpose.
Embodiment
With reference to specific embodiment, the present invention is furture elucidated.
Embodiment
The present invention provides a kind of prevention and suppressing method applied to water body cyanobacteria, comprises the following steps, step 1, first Determine the waters of application bioactivity bacteria agent product, recycle sprinkling machinery to be uniformly applied to bioactivity bacteria agent and waits to locate Waters is managed, wherein, bioactivity bacteria agent is with bacillus subtilis（Bacillus subtilis）, Pseudomonas stutzeri （Pseudomonas stutzeri）, candida aquatica（Candida aquatica）For strain, using organic and inorganic original Material, is formulated compound after each strain individually fermentation.Step 2, be monitored pending waters, when 7-8 is small after, then treating Handle waters and apply chlorella solution by spraying machinery, wherein, the chlorella content of chlorella solution is 3~8 × 107 Cfu/ml, pH value are 6.5-9.5.Step 3, to spray chlorella solution waters be monitored, when 6-7 is small after, according to monitoring The cyanobacteria state of detection, repeat step 1, the processing step of step 2.
It is further preferred that the bioactivity bacteria agent in the step 1, effective total viable count（Containing bacillus subtilis, Pseudomonas stutzeri, candida aquatica）≥2×108 A/mL, bacillus number >=1 × 108A/mL, wherein, it is effectively living Bacterium is containing bacillus subtilis, Pseudomonas stutzeri, candida aquatica；And in the step 3, before low temperature drying, treating low temperature Spray water is uniformly sprayed on the cereal of drying, sprinkling water temperature is 10 degrees Celsius -12 degrees Celsius, then natural air drying 30 divides in warehouse Clock -60 minutes；And the pH of bioactivity bacteria agent is 4-6.5 in the step 1, the sum of mould miscellaneous bacteria is≤3.0 × 105 Cfu/ml, coliform are≤10 MPN/100mL；, it is necessary to microbial inoculum when bioactivity bacteria agent sprays in the step 1 Living bacteria count is measured, and is comprised the following steps, and the sample 25mL fully mixed, be put into containing 225mL with sterile working by a 1 is made in the sterilizing conical flask of sterile biological brine：10 uniform dilution.B, 1 is drawn with 1mL sterilized straws：10 dilutions 1mL, injects in the test tube containing 9mL sterile salines, shaking mixes, and makes 1 slowly along tube wall：100 dilution.c、 1mL sterilized straws separately are taken, by aforesaid operations order, 10 times of incremental dilutions is done, is so often incremented by once, that is, uses 1 1mL instead Sterilized straw.D, 1 is selected：10-7、1：10-8Two acceptable diluent degree, respectively take 1mL to be separately added into and count culture medium plate, often A dilution factor makees three plates.E, dilution is moved into plate, should will be cooled to 45 DEG C of counting culture medium injection plate about in time 15mL-20mL, and rotate plate and make to be uniformly mixed, while culture medium will be counted and be poured into sterilizing added with 1mL dilution samples Making blank control in the sterilizing plates of physiological saline, bacillus subtilis, Pseudomonas stutzeri use dextrose culture-medium culture, Candida aquatica uses starch culture-medium culture.F, after agar completely solidification, tablet is overturn, is put in 32 DEG C of ± 1 DEG C of incubators 48h ± 2h is cultivated, tablet of the clump count between 30-300 is chosen and is counted.G, bacterium number calculates, and is adopted according to each dilution factor With the average of three flat-plate bacterial colonies, the extension rate of sample is then multiplied by, obtains viable count in every milliliter of sample；It is dilute if any two How degree of releasing, the bacterium colony of its growth to determine between 30-300 depending on ratio between two, as its ratio is less than 2, should report that it is flat Mean；Such as larger than 2, then report wherein less numeral.
The technical program, when bioactivity bacteria agent sprays in the step 1, it is necessary to the living bacteria count of microbial inoculum into Row measure, principle are to form the microorganism of bacterium colony because of its growth so that the turbidity of liquid culture increases, the sample of bacteria suspension containing bacterium The concentration of product is inversely proportional with light transmittance within the specific limits, with OD value（OD values）It is directly proportional, utilize ultraviolet specrophotometer （Photoelectric colorimetry）The OD values of the sample bacteria suspension under certain wavelength are measured, show the bacteria containing amount of sample indirectly, including it is following Step, a, will first pour into 10 mm cuvettes for cultivating the fluid nutrient medium of strain, select 600nm wavelength spectrophotometers Upper adjusting zero point, as blank control.B, 300 mL of sample is put into sterilizing wide-mouth bottle again, up and down acutely shaking 30 times -40 times, Suctioned out with sterilized straw in injection 10mm cuvettes, measure the OD value under 600nm wavelength.
The prevention applied to water body cyanobacteria of the present invention and suppressing method, the component of starch culture-medium is in the step 1 Starch 20g, potassium nitrate 1.0g, dipotassium hydrogen phosphate 0.5g, sodium chloride 0.5g, magnesium sulfate 0.5g, ferric sulfate 0.01g, add after mixing To 1000mL, pH7.2 ± 0.1, first adds all of above component in distilled water distilled water, by 2% addition agar, reheats Dissolving, corrects pH 7.2 ± 0.1, finally packing, 121 DEG C, time 15min-20min of autoclaving, heating fusing during use Agar, uses when being cooled to 45 DEG C；And glucose cultivates glucose 6g, ammonium chloride 1.0g, the sodium chloride of base in the step 1 1.0g, 1.0 g of dipotassium hydrogen phosphate, potassium dihydrogen phosphate 1.0g, magnesium sulfate 0.2g, after mixing plus distilled water is to 1000mL, and pH7.2 ± 0.1, all of above component is added in distilled water first, by 2% addition agar, dissolving is reheated, corrects pH 7.2 ± 0.1, Finally dispense, 121 DEG C of autoclaving, the time is 15min-20 min, and heating fusing agar during use, makes when being cooled to 45 DEG C With；And in the step 1 when bioactivity bacteria agent sprays, it is necessary to be measured to the mould miscellaneous bacteria of microbial inoculum, trained using Martin Base test is supported, composition is potassium dihydrogen phosphate 1.0g, glucose 10.0g, is first added all of above composition in distilled water, then add Rose-bengal aqueous solution, separately dissolves chloramphenicol with a small amount of ethanol, adds in culture medium and dissolves by heating, finally packing, autoclaving 121 DEG C, the time is 15min-20 min, and heating fusing agar during use, uses when being cooled to 45 DEG C.
The prevention applied to water body cyanobacteria of the present invention and suppressing method, when spraying chlorella solution in the step 2, need Want the chlorella content in chlorella solution to be tested, comprise the following steps, a, microscopy counting chamber, before sample-adding, first to counting The counting chamber of plate carries out microscopy, and observation counting chamber answers no-sundries.B, it is loaded, dry blood counting chamber will be cleaned and closed the lid glass Piece, drips a droplet by coverslip edge by bead algae solution with sterile pipette, allows algae solution to lean on capillary osmosis certainly along gap Counting chamber is advanced into, there can not be bubble generation.C, microscopic counting, after standing 5min, microscope is placed by blood counting chamber On objective table, counting chamber first is found with low power lens, high power lens is converted into and is counted, counting chamber is selected 5 middle lattice and counted Number.D, calculate, count the quantity A of chlorella in lattice in 5, then in algae solution chlorella quantity=A/80 × 100 × 10000 × dilute Multiple/mL is released, wherein, blood counting chamber is 25 × 16, and counting chamber thickness is 0.1mm.E, clear up, by blood counting chamber, burn Used article is cleaned out in the experiment such as cup, test tube, pipette, microscope.F, the measure of pH values, takes and pours into burning in right amount In cup, pH values are directly measured with the acidometer of calibrated mistake.
The prevention applied to water body cyanobacteria of the present invention and suppressing method, chlorella content in the chlorella solution into , it is necessary to carry out fresh water culture medium during row test（Method one）, wherein, cultivate reagent component is NaNO3、K2HPO4、MgSO4· 7H2O, CaCl22H2O, citric acid, ferric citrate or ironic citrate, Na2EDTA, Na2CO3, A5, wherein, each composition contains Amount is respectively 1.5000g/L, 0.0400g/L, 0.0750g/L, 0.0360 g/L, 0.0060g/L, 0.006g/L or 0.004g/ L、0.0010g/L、0.0200g/L、1.0mL；The A5 by ZnSO47H2O, CuSO45H2O, MoO3, H3BO3, MnCl24H2O and content are respectively 0.222g/L, 0.079g/L, 0.015g/L, 2.86g/L, 1.81g/L；And the bead , it is necessary to carry out sea water medium when chlorella content in algae solution is tested, wherein, it is made of liquid A and liquid B, solution A Agent formulations are distinguished for FeCl36H2O, MnCl24H2O, H3BO3, Na2EDTA, NaH2PO42H2O, NaNO3 and content For 6.5g, 1.8g, 168g, 225g, 100g, 500g, second liquid agent formulations are ZnCl2, CoCl26H2O, (NH4) 6Mo7O244H2O, CuSO45H2O and content are respectively 2.1g, 2.0g, 0.9g, 2.0g, and preparation method is first by solution A It is dissolved in by content sequence in 5L pure water, then second liquid is dissolved in 100ml pure water by content sequence, then solution A and second liquid is gone out Bacterium, then by the liquid A and liquid B after sterilizing, solution A：Second liquid=1000:1 or 500：1（Room temperature is kept in dark place, and placing 3~4 weeks makes With）.
The prevention applied to water body cyanobacteria of the present invention and suppressing method, chlorella content in the chlorella solution into , it is necessary to carry out fresh water culture medium during row test（Method two）, wherein, cultivate reagent component is NaNO3、K2HPO4、MgSO4· 7H2O, CaCl22H2O, citric acid, ferric citrate or ironic citrate, Na2EDTA, Na2CO3, A5, wherein, each composition contains Amount is respectively 1.5000g/L, 0.0400g/L, 0.0750g/L, 0.0360 g/L, 0.0060g/L, 0.006g/L or 0.004g/ L、0.0010g/L、0.0200g/L、1.0mL；The A5 by ZnSO47H2O, CuSO45H2O, Na2MoO4, H3BO3, MnCl24H2O and content are respectively 0.222g/L, 0.079g/L, 0.21 g/L, 2.86g/L, 1.81g/.
The prevention applied to water body cyanobacteria of the present invention and suppressing method, chlorella content in the chlorella solution into , it is necessary to carry out fresh water culture medium during row test（Method three）, wherein, cultivate reagent component is NaNO3、K2HPO4、MgSO4· 7H2O, CaCl22H2O, citric acid, ferric citrate or ironic citrate, Na2EDTA, Na2CO3, A5, wherein, each composition contains Amount is respectively 1.5000g/L, 0.0400g/L, 0.0750g/L, 0.0360 g/L, 0.0060g/L, 0.006g/L or 0.004g/ L、0.0010g/L、0.0200g/L、1.0mL；The A5 by ZnSO47H2O, CuSO45H2O, Na2MoO2H2O, H3BO3, MnCl24H2O and content are respectively 0.222g/L, 0.079g/L, 0.39g/L, 2.86g/L, 1.81g/.
The prevention applied to water body cyanobacteria of the present invention and suppressing method, pass through the bioactivity bacteria agent of high-content（Bud Spore bacillus content 10,000,000,000）, energy fast decoupled applies the organic matter in waters, while supplements application beneficial to chlorella solution, makes to have Beneficial bacteria quickly absorbs the nutriment in applied waters, so as to quickly breed, harmful algae living space is reduced high-content bud Spore bacillus decomposes organic matter, is also adjusted in synchronism and applies nitrogen in waters, phosphorus ratio, final to cause by promoting nitrogen with phosphorus, consolidating phosphorus with nitrogen N∶P ratio is normal in applied waters, so as to effectively weaken the condition of cyanobacteria production, realizes the purpose efficiently prevented and administered.
The prevention applied to water body cyanobacteria of the present invention and suppressing method, its can also according to needing to use by the following method, Apply within first day bioactivity bacteria agent, 2 mu apply one bag（1000 grams/bag）, apply within second day chlorella solution, 5 mu of applications One bottle（1000ml/ bottles）, applied once per 10-15 days.
The present invention applied to the prevention of water body cyanobacteria and the strain morphological feature of suppressing method, bacillus subtilis
Bacterium（Bacillus subtilis）, 40 × 10 times of hypothalluses of microscope are in rod-shaped, 2 μm -3 μm of length, 0.7 μm wide - 0.8 μm, gemma is column or ellipse, and middle life or near middle raw, has motility, Gram's staining is the positive, in agar plate On bacterium colony it is circular, have fold sometimes in yellow-white or shallow white, neat in edge, surface；Pseudomonas stutzeri （Pseudomonas stutzeri）, 40 × 10 times of hypothalluses of microscope are rod-shaped, can produce short side under certain condition Raw flagellum, Gram's staining are feminine gender, and the bacterium colony on agar plate is circular, in yellowish white or yellow-white, neat in edge, table Face is relatively dry；Candida aquatica（Candida aquatica）, 40 × 10 times of hypothallus forms of microscope are in rod-shaped, can be produced Mycelium is given birth to, the bacterium colony on agar plate is smooth, protuberance, circular, white or canescence, and neat in edge, surface is in sometimes Mucus shape.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art For member, without departing from the principle of the present invention, some improvement can also be made, these improvement also should be regarded as the present invention's Protection domain.

Claims (10)

  1. A kind of 1. prevention and suppressing method applied to water body cyanobacteria, it is characterised in that：Comprise the following steps,
    Step 1, determine to apply the waters of bioactivity bacteria agent product first, recycles sprinkling machinery by bioactivity bacteria agent Pending waters is uniformly applied to, wherein, bioactivity bacteria agent is with bacillus subtilis（Bacillus subtilis）、 Pseudomonas stutzeri（Pseudomonas stutzeri）, candida aquatica（Candida aquatica）For strain, use Organic and inorganic raw material, is formulated compound after each strain individually fermentation；
    Step 2, be monitored pending waters, when 7-8 is small after, then pending waters pass through spray machinery apply chlorella Solution, wherein, the chlorella content of chlorella solution is 3~8 × 107Cfu/ml, pH value are 6.5-9.5；
    Step 3, to spray chlorella solution waters be monitored, when 6-7 is small after, according to monitor and detection cyanobacteria state, weight Multiple step 1, the processing step of step 2.
  2. A kind of 2. prevention and suppressing method applied to water body cyanobacteria according to claim 1, it is characterised in that：The step Bioactivity bacteria agent in rapid 1, effective total viable count（Containing bacillus subtilis, Pseudomonas stutzeri, candida aquatica） ≥2×108 A/mL, bacillus number >=1 × 108A/mL, wherein, effective viable bacteria is containing bacillus subtilis, Amur vacation unit cell Bacterium, candida aquatica.
  3. A kind of 3. prevention and suppressing method applied to water body cyanobacteria according to claim 1 or 2, it is characterised in that：Institute The pH for stating bioactivity bacteria agent in step 1 is 4-6.5, and the sum of mould miscellaneous bacteria is≤3.0 × 105 Cfu/ml, coliform For≤10 MPN/100mL；, it is necessary to be surveyed to the living bacteria count of microbial inoculum when bioactivity bacteria agent sprays in the step 1 It is fixed, comprise the following steps,
    A, the sample 25mL fully mixed is put into the sterilizing conical flask containing 225mL sterile biological brine with sterile working and made Into 1：10 uniform dilution；
    B, 1 is drawn with 1mL sterilized straws：10 dilution 1mL, inject the test tube containing 9mL sterile salines slowly along tube wall Interior, shaking mixes, and makes 1：100 dilution；
    C, 1mL sterilized straws separately are taken, by aforesaid operations order, does 10 times of incremental dilutions, be so often incremented by once, that is, use 1 instead Branch 1mL sterilized straws；
    D, 1 is selected：10-7、1：10-8Two acceptable diluent degree, respectively take 1mL to be separately added into and count culture medium plate, each dilution Degree makees three plates；
    E, dilution is moved into plate, the counting culture medium for being cooled to 45 DEG C should be injected plate about 15mL-20mL in time, and rotate Plate makes to be uniformly mixed, while is put down the sterilizing that culture medium is poured into added with the sterile saline of 1mL dilution samples is counted Make blank control in ware, bacillus subtilis, Pseudomonas stutzeri use dextrose culture-medium culture, and candida aquatica uses Starch culture-medium culture；
    F, after agar completely solidification, tablet is overturn, puts culture 48h ± 2h in 32 DEG C of ± 1 DEG C of incubators, clump count is chosen and exists Tablet between 30-300 is counted；
    G, bacterium number calculates, the average according to each dilution factor using three flat-plate bacterial colonies, is then multiplied by the extension rate of sample, Obtain viable count in every milliliter of sample；If any two dilution factors, the bacterium colony of its growth between 30-300, depending on ratio between two how To determine, as its ratio be less than 2, should report its average；Such as larger than 2, then report wherein less numeral.
  4. A kind of 4. prevention and suppressing method applied to water body cyanobacteria according to claim 1, it is characterised in that：The step , it is necessary to be measured to the living bacteria count of microbial inoculum when bioactivity bacteria agent sprays in rapid 1, comprise the following steps,
    A, it will first pour into 10 mm cuvettes, selected on 600nm wavelength spectrophotometers for cultivating the fluid nutrient medium of strain Zero point is adjusted, as blank control；
    B, 300 mL of sample is put into sterilizing wide-mouth bottle again, acutely shaking 30 times -40 times, are suctioned out with sterilized straw and injected up and down In 10mm cuvettes, the OD value under 600nm wavelength is measured.
  5. A kind of 5. prevention and suppressing method applied to water body cyanobacteria according to claim 3, it is characterised in that：The step The component of starch culture-medium is starch 20g, potassium nitrate 1.0g, dipotassium hydrogen phosphate 0.5g, sodium chloride 0.5g, magnesium sulfate in rapid 1 0.5g, ferric sulfate 0.01g, after mixing plus distilled water is to 1000mL, and pH7.2 ± 0.1, all of above component is added distill first In water, by 2% addition agar, dissolving is reheated, corrects pH 7.2 ± 0.1, finally packing, 121 DEG C of autoclaving, the time is 15min-20min, heating fusing agar, uses when being cooled to 45 DEG C during use.
  6. A kind of 6. prevention and suppressing method applied to water body cyanobacteria according to claim 3, it is characterised in that：The step Glucose cultivates glucose 6g, ammonium chloride 1.0g, sodium chloride 1.0g, 1.0 g of dipotassium hydrogen phosphate, the potassium dihydrogen phosphate of base in rapid 1 1.0g, magnesium sulfate 0.2g, after mixing plus distilled water is to 1000mL, and pH7.2 ± 0.1, all of above component is added distill first In water, by 2% addition agar, dissolving is reheated, pH 7.2 ± 0.1 is corrected, finally dispenses, 121 DEG C of autoclaving, the time is 15min-20 min, heating fusing agar, uses when being cooled to 45 DEG C during use.
  7. 7. according to a kind of prevention and suppressing method applied to water body cyanobacteria described in claim 1, it is characterised in that：The step , it is necessary to be measured to the mould miscellaneous bacteria of microbial inoculum when bioactivity bacteria agent sprays in 1, tested using Martin's culture medium, composition For potassium dihydrogen phosphate 1.0g, glucose 10.0g, all of above composition is added in distilled water first, then adds rose-bengal water-soluble Liquid, separately dissolves chloramphenicol with a small amount of ethanol, adds in culture medium and dissolves by heating, finally packing, 121 DEG C of autoclaving, and the time is 15min-20 min, heating fusing agar, uses when being cooled to 45 DEG C during use.
  8. 8. according to a kind of prevention and suppressing method applied to water body cyanobacteria described in claim 1, it is characterised in that：The step , it is necessary to the chlorella content in chlorella solution is tested when spraying chlorella solution in 2, comprise the following steps,
    A, microscopy counting chamber, before sample-adding, first carries out microscopy, observation counting chamber answers no-sundries to the counting chamber of tally；
    B, it is loaded, dry blood counting chamber covered will be cleaned, with sterile pipette by bead algae solution by coverslip Edge drips a droplet, allows algae solution to be advanced into counting chamber certainly by capillary osmosis along gap, can not there is bubble generation；
    C, microscopic counting, after standing 5min, blood counting chamber is placed on microscope carrier, first uses low power lens
    Counting chamber is found, high power lens is converted into and is counted, counting chamber is selected 5 middle lattice and counted；
    D, calculate, count the quantity A of chlorella in lattice in 5, then in algae solution chlorella quantity=A/80 × 100 × 10000 × dilute Multiple/mL is released, wherein, blood counting chamber is 25 × 16, and counting chamber thickness is 0.1mm；
    E, clear up, used article is cleaned out during blood counting chamber, beaker, test tube, pipette, microscope etc. are tested；
    F, the measure of pH values, takes and pours into right amount in beaker, and pH values are directly measured with the acidometer of calibrated mistake.
  9. 9. according to a kind of prevention and suppressing method applied to water body cyanobacteria described in claim 8, it is characterised in that：The bead , it is necessary to carry out fresh water culture medium when chlorella content in algae solution is tested, wherein, cultivate reagent component is NaNO3、 K2HPO4, MgSO47H2O, CaCl22H2O, citric acid, ferric citrate or ironic citrate, Na2EDTA, Na2CO3, A5, Wherein, respectively composition content be respectively 1.5000g/L, 0.0400g/L, 0.0750g/L, 0.0360 g/L, 0.0060g/L, 0.006g/L or 0.004g/L, 0.0010g/L, 0.0200g/L, 1.0mL；The A5 is by ZnSO47H2O, CuSO4 5H2O, MoO3, H3BO3, MnCl24H2O and content be respectively 0.222g/L, 0.079g/L, 0.015g/L, 2.86g/L, 1.81g/L。
  10. 10. according to a kind of prevention and suppressing method applied to water body cyanobacteria described in claim 8, it is characterised in that：It is described small , it is necessary to carry out sea water medium when chlorella content in ball algae solution is tested, wherein, it is made of liquid A and liquid B, first Liquid agent formulations are FeCl36H2O, MnCl24H2O, H3BO3, Na2EDTA, NaH2PO42H2O, NaNO3 and content point Not Wei 6.5g, 1.8g, 168g, 225g, 100g, 500g, second liquid agent formulations for ZnCl2, CoCl26H2O, (NH4) 6Mo7O244H2O, CuSO45H2O and content are respectively 2.1g, 2.0g, 0.9g, 2.0g, and preparation method is first by solution A It is dissolved in by content sequence in 5L pure water, then second liquid is dissolved in 100ml pure water by content sequence, then solution A and second liquid is gone out Bacterium, then by the liquid A and liquid B after sterilizing, solution A：Second liquid=1000:1 or 500：1.
CN201711334524.4A 2017-12-14 2017-12-14 A kind of prevention and suppressing method applied to water body cyanobacteria Pending CN107986456A (en)

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