CN106630190B - Fungus agent for degrading COD and preparation method thereof - Google Patents
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Abstract
The invention provides a fungus agent for degrading COD (chemical oxygen demand) and a preparation method thereof, and the fungus agent consists of a compound fungus bacterial liquid, a protective agent and a carrier, wherein the compound fungus bacterial liquid comprises Aspergillus calidotustus bacterial liquid, Trichoderma citrinoviride bacterial liquid, Aspergillus niger bacterial liquid and Rhizopus stolonifer bacterial liquid, and the protective agent comprises skimmed milk powder, trehalose, calcium carbonate and glycerol. The preparation method comprises the following steps: 1) mixing Aspergillus calidoteus bacterial liquid, Trichoderma citrinoviride bacterial liquid, Aspergillus niger bacterial liquid and Rhizopus stolonifer bacterial liquid to obtain composite fungus bacterial liquid, and adding protective agent to mix well; 2) crushing the carrier and sieving the crushed carrier with a 60-100-mesh sieve; 3) and adding the crushed carrier into the composite fungus liquid, uniformly mixing, pre-freezing, and drying. The microbial inoculum takes fungi as degrading microorganisms, has stable strains and obvious synergistic effect, and is favorable for degrading COD in sewage.
Description
Technical Field
The invention belongs to the field of water treatment microbial preparations, and particularly relates to a fungus agent for degrading COD (chemical oxygen demand) and a preparation method thereof.
Background
At present, the treatment of industry, breed and domestic waste water plays incomparable effect in the aspect of the environmental protection, but still have a lot of problems in the sewage treatment field the inside, for example industrial sewage is unstable when handling the quality of water effect, and the operating cost is higher, water after the purification needs to carry out disinfection treatment through letting in chlorine etc. lead to originally to take place a large amount of deaths to the microorganism that will carry the inside, thereby let the water lose original ability of self-purification, there is the phenomenon on the high side in the chemical composition in the sewage treatment agent, cause secondary pollution etc. easily to the water, the COD and the ammonia nitrogen of aquaculture sewage are ultrahigh.
Compared with a chemical method, the biological method for treating the wastewater has the advantages that ⑴ each chemical product is a product with strong pertinence, the product is likely to lose efficacy when meeting other chemical substances, a biological agent has spectrum property for removing pollutants, ⑵ chemical products can temporarily eliminate certain harmful substances and cover odor, but cannot prevent the generation of the harmful substances, ⑶ after the chemical products are used, residues exist in water and can cause secondary pollution, the biological agent contains natural microorganisms without pathogenic bacteria and pathogens, the microorganisms take organic nutrients in the wastewater as food under the catalytic action of enzyme, the microorganisms are gradually reduced along with the reduction of the pollutants until the microorganisms disappear after the wastewater is purified, ⑷ is nontoxic, has no corrosivity, is convenient to use, basically does not need to add equipment or engineering, and saves capital investment.
The industrial sewage has more complex components, particularly a large amount of artificially synthesized compounds enter the environment, the substances mainly comprise alkanes, alkenes, alicyclic hydrocarbons, aromatic hydrocarbons and artificially synthesized organic substances, particularly macromolecules, difficultly degraded toxic and harmful substances such as phenol, chlorophenol, cresol, nitrophenol and the like, aromatic hydrocarbons, cyanogens, amines, ammonia nitrogen and the like enter the environment along with the waste water or solid waste discharged by a factory, and the substances cannot be decomposed and utilized by microorganisms in a short time due to the complexity of the structure of the substances and the strangeness of organisms. The traditional wastewater treatment method can not effectively remove the pollutants by culturing and domesticating microorganisms by using activated sludge, and the substances are accumulated in the environment for a long time, so that the ecological environment which depends on the survival of people is greatly polluted. The prior microbial degradation COD treatment agent has the following problems:
1) in the market, the composite microbial preparation in the COD degrading microbial inoculum generally takes bacteria as the main part, the application of the fungus composite microbial inoculum is less, and the degradation effect of the fungus is not fully excavated; 2) the problem of unstable strains seriously influences the production and application of microbial preparations, and the strains are unstable in purification and domestication; 3) the stability of the compound microbial preparation is not strong, the compound microbial preparation is easily influenced by the environment, the group structure or the change of the environment causes the death of certain microbes or the group advantage of the microbial compound bacteria agent is changed due to the intervention of new microbes; 4) the distribution of microorganisms in the compound microbial agent is not uniform enough, and the distribution of microorganisms of the same species is concentrated, so that a regionalization phenomenon is presented; 5) the research on the interrelation and the action among the bacteria in most mixed bacteria systems is not deep enough, so that the developed product is a random process for the screening and combination of the strains with a synergistic action relationship, and lacks effective theoretical guidance, and the relationship among the bacteria cannot be effectively coordinated for the applied mixed culture system, so that the optimum ecological level is achieved, and the maximum effect is exerted.
Disclosure of Invention
In order to solve the problems of the existing compound microbial preparation, the invention provides the fungus agent for degrading COD, which takes fungi as degrading microorganisms, has stable strains and obvious synergistic effect and is beneficial to degrading COD in sewage.
The technical scheme of the invention is realized as follows:
the fungus agent for degrading COD consists of a compound fungus liquid, a protective agent and a carrier, wherein the compound fungus liquid comprises Aspergillus caliidus liquid, Trichoderma citrinoviride liquid, Aspergillus niger liquid and Rhizopus stolonifer liquid, and the protective agent comprises skimmed milk powder, trehalose, calcium carbonate and glycerol.
Further, the volume ratio of the Aspergillus calidolustius bacterial liquid to the Trichoderma citrinoviride bacterial liquid to the Aspergillus niger bacterial liquid to the Rhizopus stolonifer bacterial liquid is 15-30: 10-25: 20-35: 10 to 30.
Further, the number of spores in the Aspergillus calidotustus bacterial liquid reaches 1.5-5.5 multiplied by 105cfu/ml, the number of spores of the trichoderma viride liquid reaches 1.5-5.5 multiplied by 105cfu/ml, the number of spores of the Aspergillus niger liquid reaches 1.5-5.5 multiplied by 105cfu/ml, the number of spores of the rhizopus stolonifer bacterial liquid reaches 1.5-5.5 multiplied by 105cfu/ml。
Further, the volume ratio of the mass of the skimmed milk powder to the volume of the composite fungus bacterial liquid is 5-15 g:100ml, the volume ratio of the mass of the trehalose to the volume of the composite fungus bacterial liquid is 1-10 g:100ml, the volume ratio of the mass of the calcium carbonate to the volume of the composite fungus bacterial liquid is 1-10 g:100ml, and the volume ratio of the mass of the glycerol to the volume of the composite fungus bacterial liquid is 0.5-5.5 g:100 ml.
Further, the carrier is bran or rice bran.
Further, the mass ratio of the carrier to the volume of the composite fungus liquid is 1g: 0.5-3 ml.
The invention also aims to provide a preparation method of the fungus agent for degrading COD, which comprises the following steps:
1) mixing Aspergillus calidoteus bacterial liquid, Trichoderma citrinoviride bacterial liquid, Aspergillus niger bacterial liquid and Rhizopus stolonifer bacterial liquid to obtain composite fungus bacterial liquid, and adding protective agent to mix well;
2) crushing the carrier and sieving the crushed carrier with a 60-100-mesh sieve;
3) and adding the crushed carrier into the composite fungus liquid, uniformly mixing, pre-freezing, and drying.
Preferably, in some embodiments of the invention, the Aspergillus calidolustus culture method is:
⑴ first-stage slant culture comprises culturing potato 200g, glucose 20g, magnesium sulfate 1.5g peptone 2g, potassium dihydrogen phosphate 1.5g, dipotassium hydrogen phosphate 1.5g, agar 20g, adding water to 1000ml, pH 7.0-8.0, inoculating Aspergillus calidus onto slant of the culture medium, culturing for 2-4 days, and preparing bacterial suspension containing 10% bacteria liquid5cfu/ml；
⑵ second-stage seed culture, inoculating Aspergillus caliidus to the second-stage seed culture medium according to 1% of inoculation amount, shaking for culture (rotation speed 180rpm), culturing at 23-30 deg.C for 2-4 days, diluting, coating and counting, wherein the second-stage seed culture medium comprises magnesium sulfate 0.35g, sodium chloride 0.45g, potassium dihydrogen phosphate 1.15g, urea 5.65g, glucose 5.65g, and corn flour 6g, adding water to 1L, and adjusting pH to about 2.5.
The culture methods of Trichoderma citrinoviride, Aspergillus niger and Rhizopus stolonifer are basically the same as those of Aspergillus calidolustus, except that the species are different.
Aspergillus calidoteus bacterial solution is prepared by inoculating Aspergillus calidoteus bacteria into NaCl solution.
The invention has the beneficial effects that:
1. the strain source is convenient, the strain is screened from a sewage treatment plant and purified, is identified by molecular biology means, is domesticated in sewage, and adopts various strain preservation methods such as an ultra-low temperature refrigerator and liquid nitrogen;
2. the adopted strains are easy to culture, the production process is simple, and the prepared COD degrading fungal inoculant is easy to store and transport;
3. the microorganisms selected by the product of the invention are subjected to antagonism/synergism experiments to effectively coordinate the relationship among the bacteria, so that the bacteria reach the optimal ecological level and exert the maximum ecological effect;
4. macromolecular organic matters in the sewage are efficiently degraded, the degradation capability is strong, the effect is stable when various biochemical indexes are treated, and the regularity is strong;
5. the COD fungal inoculant has strong stability, is not easily influenced by environmental change, and effectively reaches the index of the microbial composite inoculant.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a comparison graph of COD in the effluent of the microbial inoculum experiment in example 1;
FIG. 2 is a SS comparison graph of the microbial inoculum experiment effluent of example 1.
Detailed Description
Example 1
A fungus agent for degrading COD is composed of a composite fungus liquid, a protective agent and a carrier, wherein the composite fungus liquid comprises Aspergillus caliidus liquid, Trichoderma citrinoviride liquid, Aspergillus niger liquid and Rhizopus stolonifer liquid, the protective agent comprises skimmed milk powder, trehalose, calcium carbonate and glycerol, and the carrier is wheat bran. The volume ratio of Aspergillus calidotatus bacterial liquid to Trichoderma citrinoviride bacterial liquid to Aspergillus niger bacterial liquid to Rhizopus stolonifer bacterial liquid is 15: 25:20: 30.
wherein the content of the first and second substances,the number of spores in Aspergillus calidotustus bacterial liquid reaches 4.5 × 105cfu/ml, the number of spores of the Trichoderma citrinoviride bacterial liquid reaches 4.5 multiplied by 105cfu/ml, the number of spores of Aspergillus niger liquid reaches 4.5 multiplied by 105cfu/ml, the number of spores of Rhizopus stolonifer bacterial liquid reaches 4.5 multiplied by 105cfu/ml。
The preparation method comprises the following steps:
1) mixing Aspergillus calidoteus bacterial liquid, Trichoderma citrinoviride bacterial liquid, Aspergillus niger bacterial liquid and Rhizopus stolonifer bacterial liquid to obtain composite fungus bacterial liquid, and adding protective agent to mix well; wherein the mass ratio of the skimmed milk powder to the composite fungus bacterial liquid is 10g:100ml, the mass ratio of the trehalose to the composite fungus bacterial liquid is 8g:100ml, the mass ratio of the calcium carbonate to the composite fungus bacterial liquid is 8g:100ml, and the mass ratio of the glycerol to the composite fungus bacterial liquid is 4g:100 ml;
2) crushing wheat bran and sieving the crushed wheat bran with a 80-mesh sieve;
3) and adding the crushed carrier into the composite fungus liquid, uniformly mixing, pre-freezing at-20 ℃ for 8h, and drying at 10-20 Pa pressure and-45 ℃ for 10 h.
Example 2
A fungus agent for degrading COD is composed of a composite fungus liquid, a protective agent and a carrier, wherein the composite fungus liquid comprises Aspergillus caliidus liquid, Trichoderma citrinoviride liquid, Aspergillus niger liquid and Rhizopus stolonifer liquid, the protective agent comprises skimmed milk powder, trehalose, calcium carbonate and glycerol, and the carrier is wheat bran. The volume ratio of Aspergillus calidotatus bacterial liquid to Trichoderma citrinoviride bacterial liquid to Aspergillus niger bacterial liquid to Rhizopus stolonifer bacterial liquid is 30: 20:30: 20.
wherein the number of spores in Aspergillus calidotustus bacterial liquid reaches 1.5 × 105cfu/ml, the number of spores of the Trichoderma citrinoviride bacterial liquid reaches 5.5 multiplied by 105cfu/ml, the number of spores of Aspergillus niger liquid reaches 1.5 multiplied by 105cfu/ml, the number of spores of Rhizopus stolonifer bacterial liquid reaches 5.5 multiplied by 105cfu/ml。
The preparation method comprises the following steps:
1) mixing Aspergillus calidoteus bacterial liquid, Trichoderma citrinoviride bacterial liquid, Aspergillus niger bacterial liquid and Rhizopus stolonifer bacterial liquid to obtain composite fungus bacterial liquid, and adding protective agent to mix well; wherein the mass ratio of the skimmed milk powder to the composite fungus bacterial liquid is 5g:100ml, the mass ratio of the trehalose to the composite fungus bacterial liquid is 10g:100ml, the mass ratio of the calcium carbonate to the composite fungus bacterial liquid is 2g:100ml, and the mass ratio of the glycerol to the composite fungus bacterial liquid is 0.5g:100 ml;
2) crushing rice bran and sieving the crushed rice bran with a 60-mesh sieve;
3) and adding the crushed carrier into the composite fungus liquid, uniformly mixing, pre-freezing at-20 ℃ for 8h, and drying at 10-20 Pa pressure and-45 ℃ for 10 h.
Example 3
A fungus agent for degrading COD is composed of a composite fungus liquid, a protective agent and a carrier, wherein the composite fungus liquid comprises Aspergillus caliidus liquid, Trichoderma citrinoviride liquid, Aspergillus niger liquid and Rhizopus stolonifer liquid, the protective agent comprises skimmed milk powder, trehalose, calcium carbonate and glycerol, and the carrier is wheat bran. The volume ratio of Aspergillus calidotatus bacterial liquid to Trichoderma citrinoviride bacterial liquid to Aspergillus niger bacterial liquid to Rhizopus stolonifer bacterial liquid is 15: 10:35: 10.
wherein the number of spores in Aspergillus calidotustus bacterial liquid reaches 5.5 × 105cfu/ml, the number of spores of the Trichoderma citrinoviride bacterial liquid reaches 1.5 multiplied by 105cfu/ml, the number of spores of Aspergillus niger liquid reaches 5.5 multiplied by 105cfu/ml, the number of spores of Rhizopus stolonifer bacterial liquid reaches 1.5 multiplied by 105cfu/ml。
The preparation method comprises the following steps:
1) mixing Aspergillus calidoteus bacterial liquid, Trichoderma citrinoviride bacterial liquid, Aspergillus niger bacterial liquid and Rhizopus stolonifer bacterial liquid to obtain composite fungus bacterial liquid, and adding protective agent to mix well; wherein the mass ratio of the skimmed milk powder to the composite fungus bacterial liquid is 15g:100ml, the mass ratio of the trehalose to the composite fungus bacterial liquid is 1g:100ml, the mass ratio of the calcium carbonate to the composite fungus bacterial liquid is 6g:100ml, and the mass ratio of the glycerol to the composite fungus bacterial liquid is 5.5g:100 ml;
2) crushing wheat bran and sieving with a 100-mesh sieve;
3) and adding the crushed carrier into the composite fungus liquid, uniformly mixing, pre-freezing at-20 ℃ for 8h, and drying at 10-20 Pa pressure and-45 ℃ for 10 h.
Example 4
The COD degrading fungal inoculant prepared in the example 1 is taken as a typical representative, and a wastewater COD degrading experiment is carried out.
1) The raw wastewater (effluent of a primary sedimentation tank of the existing treatment process of a certain chemical plant and activated sludge from an aeration tank are used for a treatment test) is adopted and detected through two different detection standards, wherein the two methods are ① CODcr (chemical oxygen demand) colorimetric method of American Hash potassium dichromate method, organic matters in water are oxidized by potassium dichromate in a strong acid solution, and then colorimetric detection is carried out for chemical oxygen demand ② SS (American Hash detection method).
The data of the original wastewater quality are measured as follows (3 months and 1 day):
| name (R) | Aerobic influent |
| COD(mg/L) | 560 |
| SS(mg/L) | 195 |
2) The experiments were carried out in a sewage reactor using the following method:
① microbial inoculum of example 1, raw wastewater, activated sludge, urea, potassium dihydrogen phosphate and the microbial inoculum of example 1
② blank comparison experiment, raw wastewater, activated sludge, urea and potassium dihydrogen phosphate
3) The addition amount of the selngen purifying microbial inoculum is 0.05 wt% -0.5 wt%, so that the inoculation amount can obviously improve the activity of the activated sludge, improve the degradation capability of COD (chemical oxygen demand) and improve the settling property of the sludge.
4) The COD concentration and SS concentration of the experimental effluent were measured, and the measurement data were as follows (initial date: 3 months and 1 day)
As can be seen from the data in table 1, the blank control group to which the microbial inoculum of example 1 is not added has a lower number of effective microorganisms in the activated sludge, and has a limited ability to degrade COD, and macromolecular organic substances in the sewage cannot be effectively degraded, so that the effluent COD and SS are significantly higher than those of the experimental group to which the microbial inoculum of example 1 is added, and therefore, it can be concluded that the experimental group to which the microbial inoculum of example 1 is added can effectively improve the ability to degrade COD in the sewage.
Referring to FIG. 1 (obtained by comparing and plotting COD concentrations of treated wastewater), the average COD concentration of the effluent of the experimental group added with the microbial inoculum of example 1 is 106.3mg/L, the average COD concentration of the effluent of the blank control group is 137.2mg/L, and the COD of the effluent of the experimental group added with the microbial inoculum of example 1 is significantly lower than that of the blank group, which shows that the microbial inoculum of example 1 can significantly improve the capability of a system for degrading COD in wastewater.
Referring to fig. 2 (obtained by comparing and plotting the SS concentration of the experimental treatment water), it can be seen that the SS of the effluent of the experimental group added with the microbial inoculum of example 1 is much lower than that of the blank experimental group, which indicates that the addition of the microbial inoculum of example 1 can improve the settleability of the activated sludge of the system.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (7)
1. The fungus agent for degrading COD is characterized by comprising a compound fungus liquid, a protective agent and a carrier, wherein the compound fungus liquid comprises Aspergillus calidotatus liquid, Trichoderma citrinoviride liquid, Aspergillus niger liquid and Rhizopus stolonifer liquid, and the protective agent comprises skimmed milk powder, trehalose, calcium carbonate and glycerol.
2. The COD-degrading fungal inoculant according to claim 1, wherein the volume ratio of Aspergillus calidotosus liquid to Trichoderma citrinoviride liquid to Aspergillus niger liquid to Rhizopus stolonifer liquid is 15-30: 10-25: 20-35: 10 to 30.
3. The COD-degrading fungal inoculant according to claim 1 or 2, wherein the number of spores in the Aspergillus calidotosus solution is (1.5-5.5) x 105cfu/ml, the spore number of the trichoderma citrinoviride bacterial liquid reaches (1.5-5.5) multiplied by 105cfu/ml, wherein the number of spores of the Aspergillus niger liquid reaches (1.5-5.5) x 105cfu/ml, the number of spores of the rhizopus stolonifer bacterial liquid reaches (1.5-5.5) multiplied by 105cfu/ml。
4. The COD-degrading fungal inoculant according to claim 1, wherein the ratio of the mass of the skimmed milk powder to the volume of the composite fungal inoculant is 5-15 g:100ml, the ratio of the mass of the trehalose to the volume of the composite fungal inoculant is 1-10 g:100ml, the ratio of the mass of the calcium carbonate to the volume of the composite fungal inoculant is 1-10 g:100ml, and the ratio of the mass of the glycerol to the volume of the composite fungal inoculant is 0.5-5.5 g:100 ml.
5. The COD degrading fungal inoculant according to claim 1, wherein said carrier is bran or rice bran.
6. The COD-degrading fungal inoculant according to claim 1 or 2, wherein the ratio of the mass of the carrier to the volume of the composite fungal inoculum is 1g: 0.5-3 ml.
7. The method for preparing the COD-degrading fungal inoculant according to claim 1, comprising the following steps:
1) mixing Aspergillus calidoteus bacterial liquid, Trichoderma citrinoviride bacterial liquid, Aspergillus niger bacterial liquid and Rhizopus stolonifer bacterial liquid to obtain composite fungus bacterial liquid, and adding protective agent to mix well;
2) crushing the carrier and sieving the crushed carrier with a 60-100-mesh sieve;
3) and adding the crushed carrier into the composite fungus liquid, uniformly mixing, pre-freezing, and drying.
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| CN102154111A (en) * | 2010-10-28 | 2011-08-17 | 河南省科学院生物研究所有限责任公司 | Fungus bactericide for degrading organophosphorus insecticide with broad spectrum |
| CN102344899A (en) * | 2011-10-21 | 2012-02-08 | 沈阳建筑大学 | Preparation method and application for compound fungus agent for degrading organic matter |
| CN103525870A (en) * | 2013-10-23 | 2014-01-22 | 嘉兴学院 | Microbial flocculant as well as preparation method and application thereof |
| CN103540549A (en) * | 2013-10-24 | 2014-01-29 | 哈尔滨工业大学 | Preparation method of compound microbial inoculum for degrading domestic sewage |
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