CN117049710A - Treatment method for biologically purifying biogas slurry by combining microalgae-bacteria-fungi symbionts and strigolactone analogues - Google Patents
Treatment method for biologically purifying biogas slurry by combining microalgae-bacteria-fungi symbionts and strigolactone analogues Download PDFInfo
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- 239000002002 slurry Substances 0.000 title claims abstract description 76
- XHSDUVBUZOUAOQ-WJQMYRPNSA-N (3e,3ar,8bs)-3-[[(2r)-4-methyl-5-oxo-2h-furan-2-yl]oxymethylidene]-4,8b-dihydro-3ah-indeno[1,2-b]furan-2-one Chemical class O1C(=O)C(C)=C[C@@H]1O\C=C/1C(=O)O[C@@H]2C3=CC=CC=C3C[C@@H]2\1 XHSDUVBUZOUAOQ-WJQMYRPNSA-N 0.000 title claims abstract description 34
- 238000000034 method Methods 0.000 title claims abstract description 29
- 241000233866 Fungi Species 0.000 claims abstract description 27
- 241000894006 Bacteria Species 0.000 claims abstract description 19
- 238000000746 purification Methods 0.000 claims abstract description 9
- 238000012258 culturing Methods 0.000 claims description 24
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 12
- 238000005286 illumination Methods 0.000 claims description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 239000012153 distilled water Substances 0.000 claims description 8
- 239000001963 growth medium Substances 0.000 claims description 8
- 239000002609 medium Substances 0.000 claims description 8
- 241000194108 Bacillus licheniformis Species 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 5
- 229920001817 Agar Polymers 0.000 claims description 4
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 4
- 240000008397 Ganoderma lucidum Species 0.000 claims description 4
- 235000001637 Ganoderma lucidum Nutrition 0.000 claims description 4
- 240000001462 Pleurotus ostreatus Species 0.000 claims description 4
- 235000001603 Pleurotus ostreatus Nutrition 0.000 claims description 4
- 239000008272 agar Substances 0.000 claims description 4
- FRHBOQMZUOWXQL-UHFFFAOYSA-L ammonium ferric citrate Chemical compound [NH4+].[Fe+3].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O FRHBOQMZUOWXQL-UHFFFAOYSA-L 0.000 claims description 4
- 229940041514 candida albicans extract Drugs 0.000 claims description 4
- 229960004642 ferric ammonium citrate Drugs 0.000 claims description 4
- 239000004313 iron ammonium citrate Substances 0.000 claims description 4
- 235000000011 iron ammonium citrate Nutrition 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 239000011734 sodium Substances 0.000 claims description 4
- IFGCUJZIWBUILZ-UHFFFAOYSA-N sodium 2-[[2-[[hydroxy-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyphosphoryl]amino]-4-methylpentanoyl]amino]-3-(1H-indol-3-yl)propanoic acid Chemical compound [Na+].C=1NC2=CC=CC=C2C=1CC(C(O)=O)NC(=O)C(CC(C)C)NP(O)(=O)OC1OC(C)C(O)C(O)C1O IFGCUJZIWBUILZ-UHFFFAOYSA-N 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
- 239000007787 solid Substances 0.000 claims description 4
- 229910021654 trace metal Inorganic materials 0.000 claims description 4
- 239000012138 yeast extract Substances 0.000 claims description 4
- 241001185310 Symbiotes <prokaryote> Species 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 3
- 230000001580 bacterial effect Effects 0.000 claims description 2
- 239000000203 mixture Substances 0.000 claims description 2
- 238000012364 cultivation method Methods 0.000 claims 1
- 230000007613 environmental effect Effects 0.000 abstract description 2
- 230000000694 effects Effects 0.000 description 15
- 230000000052 comparative effect Effects 0.000 description 12
- 238000006243 chemical reaction Methods 0.000 description 8
- 235000015097 nutrients Nutrition 0.000 description 8
- 238000005516 engineering process Methods 0.000 description 7
- 239000000126 substance Substances 0.000 description 7
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 241000195649 Chlorella <Chlorellales> Species 0.000 description 4
- 239000002028 Biomass Substances 0.000 description 3
- 239000008367 deionised water Substances 0.000 description 3
- 229910021641 deionized water Inorganic materials 0.000 description 3
- 230000004060 metabolic process Effects 0.000 description 3
- 241000195493 Cryptophyta Species 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 229910001385 heavy metal Inorganic materials 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- QJZYHAIUNVAGQP-UHFFFAOYSA-N 3-nitrobicyclo[2.2.1]hept-5-ene-2,3-dicarboxylic acid Chemical compound C1C2C=CC1C(C(=O)O)C2(C(O)=O)[N+]([O-])=O QJZYHAIUNVAGQP-UHFFFAOYSA-N 0.000 description 1
- 235000001674 Agaricus brunnescens Nutrition 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 240000009108 Chlorella vulgaris Species 0.000 description 1
- 235000007089 Chlorella vulgaris Nutrition 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 241000222336 Ganoderma Species 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 235000007685 Pleurotus columbinus Nutrition 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- XKMRRTOUMJRJIA-UHFFFAOYSA-N ammonia nh3 Chemical compound N.N XKMRRTOUMJRJIA-UHFFFAOYSA-N 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 235000012041 food component Nutrition 0.000 description 1
- 239000004021 humic acid Substances 0.000 description 1
- 244000144972 livestock Species 0.000 description 1
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- 239000012528 membrane Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 230000000241 respiratory effect Effects 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 239000010802 sludge Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
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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
- C02F3/32—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
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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
- C12N1/12—Unicellular algae; Culture media therefor
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- C12N1/14—Fungi; Culture media therefor
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- 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
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- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2209/00—Controlling or monitoring parameters in water treatment
- C02F2209/08—Chemical Oxygen Demand [COD]; Biological Oxygen Demand [BOD]
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- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2209/00—Controlling or monitoring parameters in water treatment
- C02F2209/16—Total nitrogen (tkN-N)
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Abstract
The invention provides a treatment method for biologically purifying biogas slurry by combining microalgae-bacteria-fungi symbionts with strigolactone analogues, and belongs to the technical field of biological environmental protection. The method fully utilizes the reciprocal symbiotic relationship among microalgae, bacteria and fungi, and the constructed microalgae-bacteria-fungi symbiont has stronger growth performance and is stable, and the biogas slurry purification efficiency of the algae-bacteria-fungi symbiont can be effectively improved by the treatment method of the microalgae-bacteria-fungi symbiont combined with strigolactone analogues.
Description
Technical Field
The invention belongs to the technical field of biological environmental protection, and particularly relates to a treatment method for biological purification of biogas slurry by combining microalgae-bacteria-fungi symbionts with strigolactone analogues.
Background
Along with the gradual scale and intensive cultivation scale of livestock and poultry, the discharge amount of the cultivation excrement is increasingly huge, and a new mode for recycling the excrement is searched for to be selected inevitably. The biogas slurry is a high-pollution product brought by the biogas engineering, is rich in nitrogen, phosphorus, potassium, humic acid, active enzyme and other nutrients, can provide carbon sources, nitrogen sources and other nutritional components for microalgae growth, but if the high-concentration ammonia nitrogen, heavy metals and the like in the biogas slurry are directly used for microalgae cultivation, certain toxic effects on the microalgae are generated, and good treatment effect is difficult to achieve. Therefore, pretreatment technology of biogas slurry is receiving a great deal of attention. Most of the traditional biogas slurry treatment technologies adopt a sequencing batch activated sludge process, a biological membrane reactor and a mutual combination method, and have the problems of lower treatment efficiency, higher cost and the like. The reciprocal characteristics of the algae-bacteria symbiota in respiratory metabolism, material metabolism and the like are beneficial to the growth of the symbiota in biogas slurry, high-efficiency denitrification and dephosphorization and heavy metal removal, so that the algae-bacteria symbiota technology has good application prospect in the field of biogas slurry purification.
The legaloctone and analogues thereof (SL) can effectively improve the purification efficiency of biogas slurry and biogas in an algae co-cultivation system. The GR24 is used as the artificially synthesized strigolactone with the highest activity, has the widest application in an algae-bacteria symbiotic system, and can induce the branching of arbuscular fungus hyphae at a lower concentration level, simultaneously induce and enhance the metabolism and photosynthesis of an algae co-culture system and promote the rapid growth of symbionts.
Disclosure of Invention
The invention provides a treatment method for biologically purifying biogas slurry by combining microalgae-bacteria-fungi symbionts with strigolactone analogues, which has good removal effect on COD, TN, TP, TC in biogas slurry.
In order to achieve the above object of the present invention, the technical solution of the present invention is specifically as follows:
a method for treating biological purification biogas slurry by combining microalgae-bacteria-fungi symbionts and strigolactone analogues comprises the following steps:
step one: culturing microalgae: performing spread cultivation on microalgae by utilizing a BG-11 culture medium;
step two: culturing bacteria;
step three: pre-culturing fungi, washing, homogenizing, and domesticating and culturing homogenized fungi strains by using biogas slurry to be treated to obtain fungus-tolerant fungi strains;
step four: centrifuging the microalgae in the first step and the bacteria in the second step, and inoculating residues obtained after centrifugation into a sterile BG11-LB culture medium for culture to form microalgae-bacteria symbionts;
step five: establishment of microalgae-bacteria-fungi symbiota: adding the fungus strain obtained in the third step into an incubator of the microalgae-bacteria symbiont in the fourth step to form a microalgae-bacteria-fungus symbiotic system, and culturing the symbiont in a constant-temperature shaking table;
step six: transferring the microalgae-bacteria-fungi symbionts obtained in the step five into biogas slurry, adding strigolactone (GR 24) with different concentrations into the biogas slurry, and treating the biogas slurry to obtain treated biogas slurry.
Preferably, the steps ofThe composition of the medium BG-11 is as follows: naNO 3 1.5g L -1 、K 2 HPO 4 ·3H 2 O 0.04g L -1 、KH 2 PO 4 ·3H 2 O 0.2g L -1 0.5mg L of ethylenediamine tetraacetic acid -1 Ferric ammonium citrate 5mg L -1 Citric acid 5mg L -1 、Na 2 CO 3 0.025mg L -1 And trace metal solution 1mL L -1 。
Preferably, the step one of the expansion cultivation uses a cool white LED lamp with an illumination intensity of 200 μm -2 s -1 The light-dark ratio is set to 12 hours (day) and 12 hours (night), the culture expansion temperature is controlled to 25+/-2 ℃, and the culture expansion time is 7 days.
Preferably, the bacterial culture of step two is specifically: the bacteria were pre-incubated on a thermostatically controlled shaking table at 25.+ -. 1 ℃ for 7d (shaking table rotation speed 150 rpm), and the obtained bacteria were washed in sterile distilled water and then homogenized with 100ml of sterile distilled water.
Preferably, the bacterium of step two is bacillus licheniformis.
Preferably, the fungus in the third step is Ganoderma lucidum and oyster mushroom.
Preferably, in the fourth step, the conditions of the BG11-LB solid medium are as follows: 10g/L tryptophan, 5g/L yeast extract, 10g/L sodium chloride, 15g/L agar powder and pH of 7.0-7.2.
Preferably, the culture conditions in the fifth step are 160 rpm/25.+ -. 2 ℃ and the culture time is 2-3 days.
Preferably, the concentration of strigolactone GR24 added in the biogas slurry in the step six is 10 -9 And 10 -11 M。
Preferably, the treatment temperature in the step six is 25 ℃, and the illumination intensity is 150 mu mol.m -2 ·s -1 The light-dark time ratio is 14h light/10 h dark; the treatment time was 7d.
The beneficial effects of the invention are that
The invention provides a treatment method for biologically purifying biogas slurry by combining microalgae-bacteria-fungi symbiota with striga-lactone analogues.
The preparation method is simple and easy to operate, low in preparation cost and potential for large-scale popularization and application.
Drawings
FIG. 1 shows the COD reduction effect of biogas slurry after 7 days of reaction in different treatment modes of the invention in examples 1-2 and comparative examples 1-2;
FIG. 2 shows the effect of TN reduction in biogas slurry after 7 days of reaction in different treatment modes of examples 1-2 and comparative examples 1-2;
FIG. 3 shows the effect of reducing TP in biogas slurry after 7 days of reaction in different treatment modes of examples 1-2 and comparative examples 1-2;
FIG. 4 shows the effect of reducing TC in biogas slurry after 7 days of reaction in different treatment modes of examples 1-2 and comparative examples 1-2.
Detailed Description
A method for treating biological purification biogas slurry by combining microalgae-bacteria-fungi symbionts and strigolactone analogues comprises the following steps:
step one: culturing microalgae: performing spread cultivation on microalgae by utilizing a BG-11 culture medium; the culture conditions are preferably as follows: the spread cultivation illumination uses a cool white LED lamp with an illumination intensity of 200 mu mol m -2 s -1 Setting the light-dark ratio to 12h (day) and 12h (night), controlling the culture expansion temperature to 25+/-2 ℃ and the culture expansion time to 7 days, and manually shaking the bottle 3 times per day periodically during the culture expansion of the chlorella; the BG-11 culture medium comprises the following components: naNO 3 1.5g L -1 、K 2 HPO 4 ·3H 2 O 0.04g L -1 、KH 2 PO 4 ·3H 2 O0.2g L -1 0.5mg L of ethylenediamine tetraacetic acid -1 Ferric ammonium citrate 5mg L -1 Citric acid 5mg L -1 、Na 2 CO 3 0.025mg L -1 And trace metal solution 1mL L -1 The method comprises the steps of carrying out a first treatment on the surface of the The microalgae is free ofThe specific limitation is a method well known in the art, and is preferably Chlorella vulgaris.
Step two: culturing bacteria; the culture is specifically as follows: pre-culturing the bacteria on a constant temperature shaking table at 25+ -1deg.C for 7d (shaking table rotation speed of 150 rpm), washing the obtained bacteria in sterile distilled water, and homogenizing with 100ml sterile distilled water; the bacteria are not particularly limited, and may be those well known in the art, and are preferably Bacillus licheniformis.
Step three: the fungus is pre-cultured firstly, and the pre-culture is particularly preferably as follows: pre-culturing fungus spore inoculums on a constant-temperature shaking table for 7d (the rotation speed of a shaking table is 160 rpm) at the temperature of 25+/-1 ℃, washing with sterile deionized water, homogenizing, domesticating and culturing homogenized fungus strains by using biogas slurry to be treated, and domesticating to enable the fungus to have higher tolerance and rapid growth capacity to the biogas slurry to obtain fungus-tolerant fungus strains; the fungus is preferably Ganoderma and Pleurotus ostreatus.
Step four: centrifuging the microalgae of step one and the bacteria of step two at 8000 Xg for 3 min, inoculating the residue in sterile BG11-LB medium, and culturing at 28deg.C with light intensity of 120 μm -2 s -1 Is cultured in a sterile chamber of a microalgae-bacteria symbiont is formed; the conditions of the BG11-LB solid medium are as follows: 10g/L tryptophan, 5g/L yeast extract, 10g/L sodium chloride, 15g/L agar powder and pH of 7.0-7.2. The concentration of the microalgae is preferably 1×10 8 The concentration of bacteria is preferably 1X 10 per mL 6 cells/mL。
Step five: establishment of microalgae-bacteria-fungi symbiota: adding the fungus strain obtained in the third step into an incubator of the microalgae-bacteria symbiont in the fourth step to form a microalgae-bacteria-fungus symbiotic system, and culturing the symbiont in a constant-temperature shaking table; the concentration of the fungus strain is preferably 1.0X10 6 spores mL -1 The method comprises the steps of carrying out a first treatment on the surface of the The culture conditions are preferably 160 rpm/25.+ -. 2 ℃ and the culture time is 2-3 days.
Step six: the fifth step is carried outTransferring the microalgae-bacteria-fungi symbionts into biogas slurry, adding strigolactone (GR 24) with different concentrations into the biogas slurry, and treating the biogas slurry to obtain treated biogas slurry. The addition concentration of strigolactone GR24 in the biogas slurry is preferably 10 -9 Or 10 -11 M, more preferably 10 -9 The method comprises the steps of carrying out a first treatment on the surface of the The treatment conditions are preferably: the temperature is 25 ℃, and the illumination intensity is 150 mu mol.m -2 ·s- 1 The light-dark time ratio is 14h light/10 h dark; the treatment time was 7d.
The invention is described in further detail below with reference to the specific examples, wherein the starting materials are commercially available.
Example 1
Culturing microalgae: the chlorella is cultivated by utilizing BG-11 culture medium, and the illumination intensity of the cultivation is 200 mu mol m by utilizing cold white LED lamp -2 s -1 Setting the light-dark ratio to 12h (day) and 12h (night), controlling the culture expansion temperature to 25+/-2 ℃ and the culture expansion time to 7d, and manually shaking the bottle 3 times per day periodically during the culture expansion period of the microalgae; the BG-11 culture medium comprises the following components: naNO 3 1.5g L -1 、K 2 HPO 4 ·3H 2 O 0.04g L -1 、KH 2 PO 4 ·3H 2 O 0.2g L -1 0.5mg L of ethylenediamine tetraacetic acid -1 Ferric ammonium citrate 5mg L -1 Citric acid 5mg L -1 、Na 2 CO 3 0.025mg L -1 And trace metal solution 1mL L -1 ；
Culturing bacteria: the Bacillus licheniformis was first pre-cultured on a thermostatically controlled shaking table at 25.+ -. 1 ℃ for 7d (shaking table rotation speed 150 rpm), the resulting bacteria were washed in sterile distilled water and then homogenized with 100mL of sterile distilled water.
Cultivation of fungi: firstly, respectively pre-culturing ganoderma lucidum inoculum on a constant-temperature shaking table for 7d (the rotation speed of a shaking table is 160 rpm) under the temperature condition of 25+/-1 ℃; then, washing the fungus particles cultured for 7d by using sterile deionized water, and homogenizing the washed fungus by using the sterile deionized water (100 mL); and finally, carrying out domestication culture on the homogenized fungus strain by utilizing the biogas slurry to be treated, and domesticating to enable the fungus to have higher tolerance and rapid growth capacity on the biogas slurry, thereby finally obtaining the tolerant fungus strain.
The concentration was 1X 10 8 Chlorella (5 mL) at a concentration of 1X 10 6 The cells/mL of Bacillus licheniformis was centrifuged at 8000 Xg for 3 min. The residue obtained after centrifugation is inoculated into 150mL of sterile BG11-LB medium and irradiated at a temperature of 28℃with 120. Mu. Mol m -2 s -1 Is cultured in a sterile chamber to form a chlorella-bacillus licheniformis symbiont. The conditions of the BG11-LB solid medium are as follows: 10g/L tryptophan, 5g/L yeast extract, 10g/L sodium chloride, 15g/L agar powder and pH of 7.0-7.2.
Then 1.0X10 times 6 spores mL -1 Adding into the incubator of chlorella-Bacillus licheniformis to form chlorella-Bacillus licheniformis-Ganoderma, and culturing the symbiont in a constant temperature shaking table (160 rpm/25+ -2deg.C) for 2-3 days.
Adding Chlorella-Bacillus licheniformis-Ganoderma to biogas slurry to obtain initial total biomass (dry weight) of about 100mg/L, and adding 10 -9 M GR24, then cultured in a constant temperature, constant light sterile room for 7 days at 25deg.C with light intensity of 150μmol.m -2 ·s -1 The light-dark time ratio is 14h light/10 h dark.
Initial values of pH, COD, TN and TP in the artificially prepared biogas slurry are respectively 6.86+/-0.45, 1026.57 +/-65.11 mg.L -1 、 132.49±8.02mg·L -1 And 14.63.+ -. 1.98 mg.L -1 . After being treated by the algae-bacteria symbiotic technology, the removal rate of nutrient substances in the biogas slurry is shown in table 1. The removal rates of COD, TN, TP and TC in the biogas slurry are 85%,82%,84% and 79% respectively.
Example 2
The conditions and procedures were the same as in example 1 except that oyster mushroom was used as the fungus.
After being treated by the algae-bacteria symbiotic technology, the removal rate of nutrient substances in the biogas slurry is shown in table 1. The removal rates of COD, TN, TP and TC in the biogas slurry are 81%,78%,82% and 74% respectively.
Comparative example 1
Other culture conditions were the same as in example 1, chlorella-Bacillus licheniformis-Ganoderma was added to the biogas slurry to an initial total biomass (dry weight) of about 100mg/L, while adding 10 -11 M GR24, then culturing in a sterile room with constant temperature and constant illumination for 7d at 25deg.C and illumination intensity of 150μmol.m -2 ·s -1 The light-dark time ratio is 14h light/10 h dark. After being treated by the algae-bacteria symbiotic technology, the removal rate of nutrient substances in the biogas slurry is shown in table 1. The removal rates of COD, TN, TP and TC in the biogas slurry are 75%,73%,77% and 65% respectively.
Comparative example 2
Other culture conditions were the same as in example 1, adding Chlorella-Bacillus licheniformis-Ganoderma to biogas slurry to an initial total biomass (dry weight) of about 100mg/L, without adding GR24, and culturing in a constant temperature and constant light sterile room at 25deg.C under light intensity of 150 μmol.m for 7d -2 ·s -1 The light-dark time ratio is 14h light/10 h dark. After being treated by the algae-bacteria symbiotic technology, the removal rate of nutrient substances in the biogas slurry is shown in table 1. The removal rates of COD, TN, TP and TC in the biogas slurry are 71%,69%,73% and 59% respectively.
Comparative example 3
Table 1 Effect of the composite treatment System on removal of Main contaminants in simulated biogas slurry
FIG. 1 shows the COD reduction effect of biogas slurry after 7 days of reaction in different treatment modes of the invention in examples 1-2 and comparative examples 1-2; FIG. 2 shows the effect of TN reduction in biogas slurry after 7 days of reaction in different treatment modes of examples 1-2 and comparative examples 1-2; FIG. 3 shows the effect of reducing TP in biogas slurry after 7 days of reaction in different treatment modes of examples 1-2 and comparative examples 1-2; FIG. 4 shows the effect of reducing TC in biogas slurry after 7 days of reaction in different treatment modes of examples 1-2 and comparative examples 1-2.
As can be seen from FIGS. 1-4 and Table 1, the addition of Chlorella-Bacillus licheniformis-Ganoderma lucidum symbiotes andthe concentration of the added chlorella-bacillus licheniformis-oyster mushroom symbiont is 10 -9 After GR24 of M is added into biogas slurry, culturing is carried out for 7 days, the removal rate of COD, TN, TP and TC in the biogas slurry is above 74%, the purifying effect of nutrient substances is good, and 10% of nutrient substances are added on the basis of adding chlorella, bacillus licheniformis and ganoderma lucidum -9 The effect is best when GR24 of M. While in comparative examples 1 and 2, the addition of the chlorella-Bacillus licheniformis-Ganoderma lucidum symbiotes was 10 -11 The removal rate of COD, TN, TP and TC in biogas slurry is reduced at GR24 of M and when only chlorella-bacillus licheniformis-ganoderma lucidum symbionts are added. Therefore, the treatment method combining microalgae-bacteria-fungi symbionts and strigolactone analogue biological purification reduction has increased application potential in biogas slurry and has the GR24 concentration of 10 -9 And in the M process, the removal effect of the biogas slurry on nutrient substances in the biogas slurry is best, and the biogas slurry has great application potential.
While the invention has been described with respect to preferred embodiments, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention, and that any such changes and modifications as described in the above embodiments are intended to be within the scope of the invention.
Claims (10)
1. A method for treating biological purification biogas slurry by combining microalgae-bacteria-fungi symbionts and strigolactone analogues is characterized by comprising the following steps of:
step one: culturing microalgae: performing spread cultivation on microalgae by utilizing a BG-11 culture medium;
step two: culturing bacteria;
step three: pre-culturing fungi, washing, homogenizing, and domesticating and culturing homogenized fungi strains by using biogas slurry to be treated to obtain fungus-tolerant fungi strains;
step four: centrifuging the microalgae in the first step and the bacteria in the second step, and inoculating residues obtained after centrifugation into a sterile BG11-LB culture medium for culture to form microalgae-bacteria symbionts;
step five: establishment of microalgae-bacteria-fungi symbiota: adding the fungus strain obtained in the third step into an incubator of the microalgae-bacteria symbiont in the fourth step to form a microalgae-bacteria-fungus symbiotic system, and culturing the symbiont in a constant-temperature shaking table;
step six: transferring the microalgae-bacteria-fungi symbionts obtained in the step five into biogas slurry, adding strigolactone (GR 24) with different concentrations into the biogas slurry, and treating the biogas slurry to obtain treated biogas slurry.
2. The method for treating biological purified biogas slurry by combining microalgae-bacteria-fungi symbiota with strigolactone analogues according to claim 1, wherein the composition of the BG-11 medium in the first step is as follows: naNO 3 1.5g L -1 、K 2 HPO 4 ·3H 2 O 0.04g L -1 、KH 2 PO 4 ·3H 2 O 0.2g L -1 0.5mg L of ethylenediamine tetraacetic acid -1 Ferric ammonium citrate 5mg L -1 Citric acid 5mg L -1 、Na 2 CO 3 0.025mg L -1 And trace metal solution 1mL L -1 。
3. The method for treating biogas slurry by biological purification of microalgae-bacteria-fungi symbiota combined with strigolactone analogues according to claim 1, wherein the step one is a spread cultivation method, and the spread cultivation illumination uses a cool white LED lamp with illumination intensity of 200 mu mol m -2 s -1 The light-dark ratio is set to 12 hours (day) and 12 hours (night), the culture expansion temperature is controlled to 25+/-2 ℃, and the culture expansion time is 7 days.
4. The method for treating a biological purified biogas slurry by combining microalgae-bacteria-fungi symbiota with strigolactone analogues according to claim 1, wherein the bacterial culture in the second step is specifically: bacteria were pre-cultured on a constant temperature shaking table at a rotation speed of 150rpm for 7d at a temperature of 25.+ -. 1 ℃ and the obtained bacteria were washed in sterile distilled water and then homogenized with 100ml of sterile distilled water.
5. The method for biologically purifying biogas slurry by combining microalgae-bacteria-fungi symbiota with strigolactone analogues according to claim 1, wherein the bacteria in the second step are bacillus licheniformis.
6. The method for biologically purifying biogas slurry by combining microalgae-bacteria-fungi symbionts with strigolactone analogues according to claim 1, wherein the fungi in the third step are ganoderma lucidum and oyster mushroom.
7. The method for treating biological purified biogas slurry by combining microalgae-bacteria-fungi symbiotes with strigolactone analogues, which is characterized in that the conditions of the BG11-LB solid medium in the fourth step are as follows: 10g/L tryptophan, 5g/L yeast extract, 10g/L sodium chloride, 15g/L agar powder and pH of 7.0-7.2.
8. The method for treating a biological purified biogas slurry by combining microalgae-bacteria-fungi symbiota with strigolactone analogues according to claim 1, wherein the culture condition in the fifth step is 160 rpm/25+/-2 ℃, and the culture time is 2-3 days.
9. The method for biologically purifying biogas slurry by combining microalgae-bacteria-fungi symbionts and strigolactone analogues according to claim 1, wherein the concentration of strigolactone GR24 added in the biogas slurry in the step six is 10 -9 And 10 -11 M。
10. A microalgae-bacteria-fungi symbiont combined strigolactone analog biological net as claimed in claim 1A method for treating biogas slurry, characterized in that the treatment temperature in the step six is 25 ℃, and the illumination intensity is 150 mu mol.m -2 ·s -1 The light-dark time ratio is 14h light/10 h dark; the treatment time was 7d.
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