CN113151033B - Compound microbial preparation for inhibiting and killing blue algae and preparation method thereof - Google Patents
Compound microbial preparation for inhibiting and killing blue algae and preparation method thereof Download PDFInfo
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Abstract
The invention relates to the field of water pollution treatment, in particular to a compound microbial preparation for inhibiting and killing blue algae and a preparation method thereof; the composite microbial inoculum comprises a composite microbial inoculum, a composite carrier and deionized water, wherein the mass ratio of the composite microbial inoculum to the composite carrier to the deionized water is 3-7: 60-80: 200-400, wherein the composite microbial inoculum comprises algicidal bacteria, photosynthetic bacteria, actinomycetes, saccharomycetes and lactic acid bacteria, and the effective viable count ratio of the algicidal bacteria, the photosynthetic bacteria, the actinomycetes, the saccharomycetes and the lactic acid bacteria is 2-3: 1 to 2:2 to 3:5 to 7: 3-4, the composite carrier comprises amino acid, protease, lipase and cellulase, and the mass ratio of the amino acid to the protease to the lipase to the cellulase is 2-3. The blue algae removal amount of the compound microbial preparation 4d reaches 100%, the blue algae removal time is shorter, the efficiency is higher, the self-cleaning capacity of a water body is improved, and the growth of aquatic organisms is promoted.
Description
Technical Field
The invention relates to the field of water pollution treatment, in particular to a compound microbial preparation for inhibiting and killing blue algae and a preparation method thereof.
Background
The eutrophication of the water body causes the mass propagation of algae, which causes the frequent outbreak of water bloom and red tide, which not only aggravates the pollution of the water body and the shortage of water resources and destroys the balance of the aquatic ecosystem, but also poses serious threat to the safety of drinking water because the bloom-forming cyanobacteria can generate and release various algal toxins mainly comprising microcystins. At present, there are various physical, chemical and biological methods for treating cyanobacterial bloom, although chemical algicides such as heavy metal copper can be used for killing cyanobacterial, natural flocculants such as iron or aluminum chemical flocculants and clay minerals can be used for rapidly settling cyanobacterial, nitrogen and phosphorus in water can support the growth of cyanobacterial to cause cyanobacterial bloom again, and chemical substance residues can cause secondary pollution to water.
The algicidal microorganism is used as an important component of a population mechanism and a function of an aquatic ecosystem, and has a very important effect on maintaining the balance of algae biomass. Compared with the traditional physical and chemical algae control methods, the microbial algae control method has the advantages of high efficiency, safety, durability and the like, and some algae-dissolving bacteria have the effects of regulating and controlling water quality, degrading harmful substances, promoting the growth of excellent microalgae and the like, so that the microbial algae control method is widely concerned. The algae-lysing microorganism lyses algae mainly through two ways: firstly, direct algae lysis is realized, namely algae bacteria directly contact with and even invade algae cells to perform algae lysis; the other is indirect algae lysis, namely algae cell lysis by secreting certain extracellular active substances or algae growth inhibition by competing nutrition with algae, wherein the indirect algae lysis by secreting proteins, polypeptides, amino acids, antibiotics, alkaloids and other ways is the main action mode of microbial algae lysis. In recent years, many microorganisms have been discovered at home and abroad to dissolve and inhibit the growth of algae in different ways. Common are mainly algal viruses, algicidal bacteria (pseudomonas, alteromonas, vibrio, bacillus, etc.), algicidal actinomycetes, protozoa, and the like. The reported microbial agents, such as microbial flora EM, liquid clearness (ACF 32), micro-Bac fermentation broth (composite microorganism) and the like, show good algae removal effect in the application of small eutrophic water surfaces and water bodies, have simple process, and provide huge expansion space for the research and application of the microorganism algae removal technology due to the situation of 'treating algae with bacteria'.
Therefore, a compound microbial preparation for inhibiting and killing blue algae and a preparation method thereof are provided.
Disclosure of Invention
The invention aims to provide a compound microbial preparation for inhibiting and killing blue algae and a preparation method thereof, the blue algae removal of the compound microbial preparation 4d reaches 100%, the time for removing the blue algae is shorter, the efficiency is higher, the self-cleaning capability of a water body is improved, and the growth of aquatic organisms is promoted.
In order to achieve the purpose, the technical scheme of the invention is as follows:
the compound microbial preparation for inhibiting and killing blue algae comprises a compound microbial agent, a compound carrier and deionized water, wherein the mass ratio of the compound microbial agent to the compound carrier to the deionized water is (3-7): 60-80: 200-400, wherein the composite microbial inoculum comprises algicidal bacteria, photosynthetic bacteria, actinomycetes, saccharomycetes and lactic acid bacteria, and the effective viable count ratio of the algicidal bacteria, the photosynthetic bacteria, the actinomycetes, the saccharomycetes and the lactic acid bacteria is 2-3: 1 to 2:2 to 3:5 to 7: 3-4, the composite carrier comprises amino acid, protease, lipase and cellulase, and the mass ratio of the amino acid to the protease to the lipase to the cellulase is 2-3.
Specifically, the algae-lysing bacteria are marine bacteria pseudoalteromonas.
Specifically, the photosynthetic bacteria are cyanobacteria.
Specifically, the actinomycetes is micromonospora.
Specifically, the yeast is Rhodotorula rubra.
Specifically, the lactobacillus is lactobacillus plantarum.
A preparation method of a compound microbial preparation for inhibiting and killing blue algae comprises the steps of weighing corresponding raw material components according to a ratio, adding amino acid, protease, lipase and cellulase into deionized water, and uniformly stirring to prepare a compound carrier suspension A; inoculating the domesticated complex microbial inoculum into the complex carrier suspension A, culturing for 5-7 d at normal temperature to prepare a bacterial suspension B, enabling the OD680 value of the bacterial suspension B to be 0.7, and drying and granulating the bacterial suspension B by using a spray dryer to prepare the complex microbial preparation with the average particle size of 40 microns.
The invention has the beneficial effects that:
(1) The raw materials of the composite carrier, namely amino acid, protease, lipase and cellulase, can be naturally degraded, so that the composite carrier is low in cost, harmless to the environment, environment-friendly and energy-saving, and can be widely applied to the repair and treatment of industrial wastewater, domestic sewage and underground water;
(2) The invention adopts various strains with the function of killing algae, is domesticated, attached with a composite carrier and dried to prepare a powdery microbial inoculum, can effectively control the biomass of the blue algae and avoid secondary pollution to natural water;
(3) The composite microbial preparation 4d provided by the invention has the advantages that the blue algae removal amount reaches 100%, the blue algae removal time is shorter, the efficiency is higher, the self-cleaning capacity of the water body is improved, the growth of aquatic organisms is promoted, the composite microbial preparation is suitable for urban river water pollution treatment and restoration, is also suitable for restoration of industrial polluted closed water bodies, can improve the micro-ecological system of the water body, and restores the biological diversity of the water body.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be obtained by a person skilled in the art without making any creative effort based on the embodiments in the present invention, belong to the protection scope of the present invention.
The compound microbial preparation for inhibiting and killing blue algae comprises a compound microbial agent, a compound carrier and deionized water, wherein the mass ratio of the compound microbial agent to the compound carrier to the deionized water is (3-7): 60-80: 200-400, wherein the composite microbial inoculum comprises algicidal bacteria, photosynthetic bacteria, actinomycetes, saccharomycetes and lactic acid bacteria, and the effective viable count ratio of the algicidal bacteria, the photosynthetic bacteria, the actinomycetes, the saccharomycetes and the lactic acid bacteria is 2-3: 1 to 2:2 to 3:5 to 7: 3-4, the composite carrier comprises amino acid, protease, lipase and cellulase, and the mass ratio of the amino acid to the protease to the lipase to the cellulase is 2-3.
Furthermore, the algae-lysing bacteria related to the invention are marine bacteria pseudoalteromonas, which are strains deposited in the common microorganism center of the China Committee for culture Collection of microorganisms (China university of oceans) as described in the China patent CN102304484B, and the preservation numbers are as follows: CGMCC NO.5009.
Further, the photosynthetic bacteria related to the present invention are cyanobacteria, the cyanobacteria is a strain deposited in the common microorganism center of the China Committee for culture Collection of microorganisms (national institute of biological energy and Process), as described in China patent CN108977371A, and the preservation number is: CGMCC No.13785.
Further, the actinomycetes of the present invention are micromonospora, which is purchased from the effective company of biological technology in Shanghai, tongpai, product number: ATCC (American ginseng).
Further, the yeast related to the invention is rhodotorula rubra purchased from Shanghai Xuan Yao Biotech limited, product number: XY-WSW-2775.
Further, the lactobacillus involved in the invention is lactobacillus plantarum, which is purchased from Weifang Ruichi Biotechnology Limited, product number: 308084-36-8.
Example 1
Referring to tables 1, 2 and 3, the scheme a in table 1 is adopted as the scheme for the proportion configuration of the effective viable count of each strain in the composite microbial inoculum, the scheme a in table 3 is adopted as the scheme for the mass ratio configuration of each component in the composite carrier, the scheme a in table 2 is adopted as the scheme for the mass portion configuration of the composite microbial inoculum, the composite carrier and deionized water, and the preparation process is as follows:
weighing corresponding raw material components according to a ratio, adding amino acid, protease, lipase and cellulase into deionized water, and uniformly stirring to prepare a composite carrier suspension A; inoculating the domesticated complex microbial inoculum into the complex carrier suspension A, culturing for 5-7 d at normal temperature to prepare a bacterial suspension B, enabling the OD680 value of the bacterial suspension B to be 0.7, and drying and granulating the bacterial suspension B by using a spray dryer to prepare the complex microbial preparation with the average particle size of 40 microns.
Example 2
The scheme B in the table 1 is adopted as the scheme for the proportion configuration of the effective viable count of each strain in the composite microbial inoculum, the scheme B in the table 3 is adopted as the scheme for the mass ratio configuration of each component in the composite carrier, the scheme B in the table 2 is adopted as the scheme for the mass part configuration of the composite microbial inoculum, the composite carrier and the deionized water, and the preparation process is the same as that in the example 1.
Example 3
The scheme C in Table 1 is adopted as the scheme for the proportion of the effective viable count of each strain in the composite microbial inoculum, the scheme C in Table 3 is adopted as the scheme for the proportion of the mass of each component in the composite carrier, the scheme C in Table 2 is adopted as the scheme for the preparation of the composite microbial inoculum, the composite carrier and the deionized water in parts by mass, and the preparation process is the same as that in example 1.
Example 4
The scheme D in the table 1 is adopted as the scheme for the proportion configuration of the effective viable count of each strain in the composite microbial inoculum, the scheme D in the table 3 is adopted as the scheme for the mass ratio configuration of each component in the composite carrier, and the scheme D in the table 2 is adopted as the scheme for the mass parts configuration of the composite microbial inoculum, the composite carrier and the deionized water, and the preparation process is the same as that in the example 1.
Comparative example 1
The scheme A in Table 1 is adopted as the scheme for the proportion of the effective viable count of each strain in the composite microbial inoculum, the scheme a in Table 3 is adopted as the scheme for the proportion of the mass of each component in the composite carrier, the scheme V in Table 2 is adopted as the scheme for the preparation of the composite microbial inoculum, the composite carrier and the deionized water, and the preparation process is the same as that in example 1.
Comparative example 2
The scheme A in Table 1 is adopted as the scheme for the proportion of the effective viable count of each strain in the composite microbial inoculum, the scheme a in Table 3 is adopted as the scheme for the proportion of the mass of each component in the composite carrier, the scheme I in Table 2 is adopted as the scheme for the preparation of the composite microbial inoculum, the composite carrier and the deionized water in parts by mass, and the preparation process is the same as that in example 1.
Comparative example 3
The scheme E in the table 1 is adopted as the scheme for the proportion configuration of the effective viable count of each strain in the composite microbial inoculum, the scheme a in the table 3 is adopted as the scheme for the mass ratio configuration of each component in the composite carrier, the scheme A in the table 2 is adopted as the scheme for the mass parts configuration of the composite microbial inoculum, the composite carrier and the deionized water, and the preparation process is the same as that in the example 1.
Comparative example 4
The scheme F in the table 1 is adopted as the scheme for the proportion of the effective viable count of each strain in the composite microbial inoculum, the scheme a in the table 3 is adopted as the scheme for the proportion of the mass of each component in the composite carrier, the scheme A in the table 2 is adopted as the scheme for the mass of the composite microbial inoculum, the composite carrier and the deionized water, and the preparation process is the same as that in the example 1.
Comparative example 5
The scheme A in Table 1 is adopted as the scheme for the proportion of the effective viable count of each strain in the composite microbial inoculum, the scheme e in Table 3 is adopted as the scheme for the proportion of the mass of each component in the composite carrier, the scheme A in Table 2 is adopted as the scheme for the preparation of the composite microbial inoculum, the composite carrier and the deionized water in parts by mass, and the preparation process is the same as that in example 1.
Comparative example 6
The scheme A in the table 1 is adopted as the scheme for the proportion configuration of the effective viable count of each strain in the composite microbial inoculum, the scheme f in the table 3 is adopted as the scheme for the mass ratio configuration of each component in the composite carrier, the scheme A in the table 2 is adopted as the scheme for the mass parts configuration of the composite microbial inoculum, the composite carrier and the deionized water, and the preparation process is the same as that in the example 1.
TABLE 1 proportion scheme of effective viable count of each strain in composite microbial inoculum
| Each strain in the composite microbial inoculum | Scheme A | Scheme B | Scheme C | Scheme D | Scheme E | Scheme F |
| Pseudoalteromonas as marine bacterium | 2 | 3 | 2.2 | 2.6 | 1.5 | 3.3 |
| Cyanobacteria | 1 | 2 | 1.5 | 1.8 | 0.8 | 2.5 |
| Micromonospora species | 2 | 3 | 2.4 | 2.9 | 1.7 | 3.2 |
| Rhodotorula rubra | 5 | 7 | 2.9 | 6.5 | 4.5 | 7.4 |
| Lactobacillus plantarum | 3 | 4 | 3.1 | 3.7 | 2 | 4.3 |
TABLE 2 preparation of composite bacterial agent, composite carrier and deionized water in parts by weight
| Scheme A | Scheme B | Scheme C | Scheme D | Scheme E | Scheme already | |
| Complex microbial inoculum | 3 | 7 | 4 | 6 | 2 | 8 |
| Composite carrier | 60 | 80 | 65 | 75 | 55 | 85 |
| Deionized water | 200 | 400 | 250 | 360 | 150 | 430 |
TABLE 3 scheme for configuring mass ratio of each component in composite carrier
| Scheme a | Scheme b | Scheme c | Scheme d | Scheme e | Scheme f | |
| Amino acids | 2 | 3 | 2.2 | 2.7 | 1.8 | 3.5 |
| Protease enzyme | 0.5 | 0.8 | 0.6 | 0.7 | 0.4 | 0.9 |
| Lipase enzyme | 0.7 | 1 | 0.8 | 0.9 | 0.6 | 1.2 |
| Cellulase enzymes | 0.3 | 0.5 | 0.38 | 0.46 | 0.2 | 0.6 |
The strain forms of examples 1 to 4 and comparative examples 1 to 6
By morphological observation of the strains, the results are as follows:
morphological characteristics of colonies in examples 1 to 4: the colony is round and small, all short hyphae are on the hypha cluster, obvious vacuoles are formed, many short fragments exist, and the colony can grow at the pH value of 5.5-6.5.
Morphological characteristics of colonies in examples 1 to 6: the colony is round and small, and the mycelium mass is distributed with vacuoles all over, and the vacuoles are large and can grow at pH5.5-6.5.
The algae lysing experiments of the above examples 1 to 4 and comparative examples 1 to 6 were carried out, wherein the used algae solution was a laboratory microcystis aeruginosa pure culture solution, and the algae seed was microcystis aeruginosa FACHB-905 purchased from a freshwater algae seed bank of Wuhan aquatic organism research institute of Chinese academy of sciences, so that the change of chlorophyll a concentration can well represent the removal effect of microcystis aeruginosa.
Sampling and determining that the initial chlorophyll a concentration of 100ml is 68.6mg/m, and placing at 28 ℃, the light intensity is 2500lux, the light circulation is 12h: standing and culturing in an illumination incubator for 12h, and sampling every 24h to determine the chlorophyll a concentration; according to the mass ratio of the composite microbial preparation to the algae liquid of 0.5:1 input, and the removal rate of the leaf green a is shown in Table 4.
TABLE 4 leaf Green a removal Rate
As can be seen from table 4 above, in the algae-lysing experiments performed in examples 1 to 4 and comparative examples 1 to 6, the blue algae removal amount of the composite microbial preparation 4d added in the formulations of examples 1 to 4 reaches 100%, which is beneficial to promoting the establishment of algae-bacteria balance in the water body, improving the self-cleaning capability of the water body, and promoting the growth of aquatic organisms; compared with the compound microbial preparation prepared by the formula of the comparative examples 1-6, the time for removing the blue algae is shorter, and the efficiency is higher.
Finally, although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that various changes and modifications may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (2)
1. The compound microbial preparation for inhibiting and killing blue algae is characterized by comprising a compound microbial agent, a compound carrier and deionized water, wherein the compound microbial agent comprises algae-lysing bacteria, photosynthetic bacteria, actinomycetes, saccharomycetes and lactic acid bacteria, and the compound carrier comprises amino acid, protease, lipase and cellulase; the mass ratio of the composite bacterial agent to the composite carrier to the deionized water is 8:85:430, the ratio of the effective viable count of the algicidal bacteria, the photosynthetic bacteria, the actinomycetes, the yeasts and the lactic acid bacteria is 2:1:2:5:3, the mass ratio of the amino acid to the protease to the lipase to the cellulase is 2.5; the algae-lysing bacteria are marine bacteria pseudoalteromonas; the photosynthetic bacteria are cyanobacteria; the actinomycetes are micromonospora; the yeast is rhodotorula rubra; the lactobacillus is lactobacillus plantarum.
2. The method for preparing a composite microbial preparation for inhibiting and killing blue-green algae according to claim 1, wherein the corresponding raw material components are weighed according to a ratio, amino acid, protease, lipase and cellulase are added into deionized water, and the mixture is uniformly stirred to prepare a composite carrier suspension A; inoculating the domesticated complex microbial inoculum into the complex carrier suspension A, culturing for 5-7 d at normal temperature to prepare a bacterial suspension B, enabling the OD680 value of the bacterial suspension B to be 0.7, and drying and granulating the bacterial suspension B by using a spray dryer to prepare the complex microbial preparation with the average particle size of 40 microns.
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