CN112931501B - Slow-release algae-inhibiting microsphere and preparation method and application thereof - Google Patents

Slow-release algae-inhibiting microsphere and preparation method and application thereof Download PDF

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CN112931501B
CN112931501B CN202110172835.5A CN202110172835A CN112931501B CN 112931501 B CN112931501 B CN 112931501B CN 202110172835 A CN202110172835 A CN 202110172835A CN 112931501 B CN112931501 B CN 112931501B
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luteolin
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李洁明
李碧滢
安广琪
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China Agricultural University
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Abstract

The invention provides a sustained-release algae-inhibiting microsphere and a preparation method and application thereof. The slow-release algae-inhibiting microsphere is prepared from the following raw materials: 1 to 1.5 percent (w/v) of luteolin, 18 to 20 percent (w/v) of olive oil, 10 to 12 percent (w/v) of emulsifier, 10 to 12 percent (w/v) of co-emulsifier, 1 to 2.5 percent (w/v) of sodium alginate, 0.2 to 0.3 percent (w/v) of chitosan, 4 to 8 percent (w/v) of calcium chloride and 0.5 to 1 percent (v/v) of cross-linking agent. The slow-release algae-inhibiting microsphere provided by the invention can slow down and control the release rate of luteolin, has the advantages of simple preparation process, low cost and no pollution to the environment, can obviously and efficiently inhibit the growth of microcystins within 30-50 days through the sustained slow release of the luteolin, and simultaneously efficiently inhibits the release of Microcystins (MCs) without repeatedly adding an algae inhibitor, and has long-term and stable effect on efficiently inhibiting the outbreak of cyanobacterial bloom.

Description

Slow-release algae-inhibiting microsphere and preparation method and application thereof
Technical Field
The invention relates to the technical field of water body environmental protection, in particular to a slow-release algae inhibiting microsphere and a preparation method and application thereof.
Background
In recent years, the cyanobacterial blooms in many large lakes and reservoirs frequently occur, and the microcystis is mostly used as a water bloom dominant species, which can destroy aquatic ecosystems by blocking sunlight and consuming oxygen. The microcystis usually also produces and releases microcystins to further pollute water bodies and poison aquatic organisms and human beings, wherein Microcystins (MCs) are the most representative. The world health organization recommends setting the content of MCs in drinking water to 1 μ g/L, and therefore controlling the growth of harmful algae and mitigating the release of MCs becomes a problem of widespread concern.
The treatment of harmful algae can be divided into physical, chemical and biological methods. However, the water treatment processes such as coagulation, precipitation and filtration are difficult to effectively remove algae, and secondary pollution is easily caused by adding chemical methods such as copper and chlorine algaecide, and MCs are released again after algae cells are broken. In contrast, the plant secondary metabolite (phytochemoattractant) can effectively inhibit the growth of microcystis and effectively remove MCs.
The slow release technology of the algistat is to store effective algistatic components in slow release microspheres (CRM) by using a physicochemical means, and then slowly and continuously release the effective algistatic components, and the preparation and the algistatic effect of the conventional CRM have the following defects: the production process is complex and the production cost is high; the algae growth inhibition effect cannot reach long-term and stable effect, and the effect needs to be further enhanced; the long-term and high-efficiency inhibition effect on the release of microcystin MCs is lacked while the growth of algae is inhibited; there is no report of CRM algae inhibiting and MCs inhibiting effect under different environmental conditions.
Disclosure of Invention
The embodiment of the invention provides a slow-release algae inhibiting microsphere which has no pollution to the environment, can exert high-efficiency, lasting and stable algae inhibiting effect under different environmental conditions, and has cheap and simple production process.
The embodiment of the invention provides a sustained-release algae inhibiting microsphere which is prepared from the following raw materials:
Figure BDA0002939315160000021
according to the invention, in the preferable selection of the luteolin, the olive oil, the emulsifier, the co-emulsifier and the sodium alginate, the denominator v is the volume of the mixed solution of the luteolin emulsifying system and the sodium alginate (in the step 1) of the preparation method of the slow-release algae-inhibiting microspheres); preferably, in the chitosan and the calcium chloride, the denominator v is the volume of the chitosan and calcium chloride aqueous solution (in the step 2) of the preparation method of the slow-release algae-inhibiting microsphere). Preferably selecting the cross-linking agent, wherein the denominator v is the total volume of the luteolin emulsifying system and the sodium alginate mixed solution added with the chitosan and calcium chloride aqueous solution (in the step 2) of the preparation method of the slow-release algae-inhibiting microspheres).
The luteolin sustained-release algae-inhibiting microsphere provided by the invention takes the plant-borne allelopathic substance luteolin (3 ', 4', 5, 7-tetrahydroxyflavone) as a core material, and the luteolin is a representative flavone class algae-inhibiting substance produced by a plurality of terrestrial and aquatic plants; chitosan and sodium alginate are used as wall materials. The invention can slow and control the release rate of luteolin; through the sustained and slow release of the luteolin, the growth of the microcystis can be obviously inhibited within 30-50 days, the repeated addition of the algistat is not needed, and the long-term and stable effect on efficiently inhibiting the outbreak of the cyanobacterial bloom is achieved. According to the invention, olive oil is used as an oil phase, TWEEN-80 is used as an emulsifier, methanol is used as an auxiliary emulsifier, a uniform luteolin emulsifying system can be obtained, sodium alginate and the luteolin emulsifying system are mixed, and calcium chloride is dripped into a chitosan solution, so that the uniform slow-release algae-inhibiting microspheres with high embedding efficiency are constructed. Sodium alginate and chitosan are used as wall materials, so that the release rate of luteolin can be slowed and controlled, and the constructed microspheres have higher structural strength.
According to the sustained-release algal inhibition microsphere provided by the embodiment of the invention, the emulsifier is selected from TWEEN, preferably TWEEN-80; and/or the coemulsifier is selected from lower alcohols, preferably methanol. In the invention, the TWEEN-80 is used as an emulsifier and the methanol is used as an auxiliary emulsifier, so that a uniform luteolin emulsifying system can be well formed, and the emulsifying system can be fully mixed with the sodium alginate, thereby uniformly distributing the luteolin in the constructed sustained-release microspheres.
According to the slow-release algae inhibiting microsphere provided by the embodiment of the invention, the cross-linking agent is glutaraldehyde. In the invention, glutaraldehyde is used as a cross-linking agent, so that the crosslinking degree of the microspheres can be effectively improved.
The slow-release algae inhibiting microsphere provided by the embodiment of the invention comprises:
Figure BDA0002939315160000031
preferably, the denominator v (luteolin, olive oil, TWEEN-80, methanol and sodium alginate) is the volume of the mixed solution of the luteolin emulsifying system and the sodium alginate in the step 1) of the preparation method of the sustained-release algae-inhibiting microspheres;
0.2% -0.3% (w/v) of chitosan
Calcium chloride 4% -8% (w/v)
Preferably, the denominator v (of chitosan and calcium chloride) is the volume of the chitosan and calcium chloride aqueous solution (in the step 2) of the preparation method of the slow-release algae-inhibiting microsphere);
glutaraldehyde 0.5% -1% (v/v)
Preferably, the (glutaraldehyde) denominator v is the total volume of the luteolin emulsifying system and the sodium alginate mixed solution added with the chitosan and calcium chloride aqueous solution (in the step 2) of the preparation method of the slow-release algae inhibiting microspheres).
According to the invention, the luteolin is used as the core material, the matrix is used as the plant growth allelochemical substance, the outbreak of the blue algae bloom can be well inhibited, and meanwhile, the slow release microspheres can ensure that the luteolin is slowly released, so that the algae inhibiting effect is effectively improved, and the algae inhibiting action time is prolonged. The luteolin is uniformly distributed in the constructed microspheres through an emulsification system, and meanwhile, the crosslinking agent glutaraldehyde can enhance the crosslinking degree of each component in the microspheres, so that the structure is more compact, and the slow release effect is enhanced.
The invention also provides a preparation method of the slow-release algae inhibiting microsphere, which comprises the following steps:
1) dissolving luteolin in olive oil, sequentially adding a co-emulsifier and an emulsifier, uniformly mixing, performing ultrasonic treatment to form a uniform luteolin emulsifying system, adding the luteolin emulsifying system into a sodium alginate solution, and stirring while fully mixing to obtain a mixed solution;
2) and dropwise adding the mixed solution into an aqueous solution of chitosan and calcium chloride to form microspheres, uniformly shaking while dropwise adding, adding a crosslinking agent, stirring, and performing aftertreatment.
According to the preparation method of the slow-release algae inhibiting microspheres provided by the embodiment of the invention, in the step 1), in the ultrasonic treatment, the ultrasonic frequency is preferably 150W; the stirring speed is 100-200 r/min, and the stirring time is 0.5-1 min.
According to the preparation method of the slow-release algae inhibiting microsphere provided by the embodiment of the invention, in the step 1), the preparation of the sodium alginate solution comprises the following steps: mixing sodium alginate powder with water, preferably heating with alcohol lamp flame until sodium alginate and water are mixed and dissolved (transparent gel).
According to the preparation method of the slow-release algae inhibiting microspheres provided by the embodiment of the invention, in the step 2), the stirring speed is 100-200 r/min, and the stirring time is determined according to the amount of the prepared microspheres; preferably, the mixed solution is slowly dripped into an aqueous solution containing chitosan and calcium chloride at a position which is 10-20 cm higher than the liquid surface, preferably 15cm higher than the liquid surface.
According to the preparation method of the slow-release algae-inhibiting microsphere provided by the embodiment of the invention, the post-treatment comprises the following steps: washing, filtering, drying and sterilizing the microspheres; preferably, the microspheres are washed with petroleum ether for 2-5 times, preferably 3 times, and then washed with distilled water for 2-5 times, preferably 3 times, and then subjected to vacuum filtration, air drying at room temperature and ultraviolet sterilization to obtain the microsphere.
In the embodiment of the invention, the embedding effect of the luteolin can be better ensured by controlling the parameters of the method, and the embedding efficiency can be further improved.
The embodiment of the invention also provides application of the slow-release algae inhibiting microspheres or the slow-release algae inhibiting microspheres prepared by the preparation method in long-acting prevention and control of algae, preferably cyanobacterial bloom.
The invention has the beneficial effects that: according to the slow-release algae-inhibiting microsphere provided by the invention, the plant secondary metabolite luteolin can effectively inhibit the growth of microcystis and effectively remove MCs, but under a natural environment, the luteolin is directly applied to harmful cyanobacterial bloom and can not maintain the persistence of an algae-resisting effect. The slow-release algae inhibiting microsphere simulates the natural release process of the phytochemical allelochemicals, thereby greatly prolonging the action time of the allelochemicals and improving the algae inhibiting effect. And has the advantages of simple preparation process, low cost, no pollution to the environment and the like, and is mainly used for preventing and treating the harmful cyanobacterial bloom with durability, high efficiency and safety. The slow-release algae-inhibiting microsphere is a spherical solid and can be put into a water body in actual use.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be clearly and completely described below, and it should be apparent that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications. The instruments and the like are conventional products which are purchased by normal distributors and are not indicated by manufacturers. The process is conventional unless otherwise specified, and the starting materials are commercially available from the open literature.
In the following examples of the present invention, the preparation of the sodium alginate solution comprises: mixing sodium alginate powder with water, and heating with alcohol lamp outer flame until sodium alginate and water are completely mixed and dissolved (transparent gel).
Example 1
The embodiment provides a sustained-release algae-inhibiting microsphere, which comprises the following raw materials:
Figure BDA0002939315160000061
note: the denominator v is the volume of the luteolin emulsifying system and sodium alginate mixed solution in the preparation method of the sustained-release algae-inhibiting microspheres.
Chitosan 0.25% (w/v)
Calcium chloride 4% (w/v)
Note: the denominator v (chitosan and calcium chloride) is the volume of the aqueous solution of the chitosan and the calcium chloride in the preparation method of the slow-release algae-inhibiting microsphere.
0.5 percent of glutaraldehyde (v/v, the denominator v is the total volume of the mixed solution of luteolin and sodium alginate after the mixed solution is dripped into the aqueous solution of chitosan and calcium chloride) in the preparation method of the slow-release algae-inhibiting microspheres.
The preparation method of the sustained-release algal inhibition microsphere provided by the embodiment is as follows:
1) emulsification: dissolving luteolin in olive oil, adding co-emulsifier methanol, adding emulsifier TWEEN-80, mixing, and performing ultrasonic treatment at 150W to form a uniform luteolin emulsifying system;
2) microsphere formation: adding luteolin emulsifying system into sodium alginate solution, stirring at 150r/min for 1min while adding until mixing completely to obtain mixed solution. Slowly dropping a mixed solution of luteolin and sodium alginate into an aqueous solution containing chitosan and calcium chloride at a position 15cm away from the water surface, slowly dropping glutaraldehyde serving as a cross-linking agent into the solution, and stirring the solution at a stirring speed of 150r/min during the dropping process to uniformly mix the solution (the stirring time is determined according to the amount of the microspheres constructed);
3) and (3) suction filtration: washing the microspheres constructed in the step 2) with petroleum ether for three times, then washing with distilled water for three times, and then carrying out vacuum filtration to obtain wet luteolin slow-release algae-inhibiting microspheres;
4) and (3) drying: placing the wet luteolin sustained-release algae-inhibiting microspheres on 0.45-micron filter paper, placing the filter paper in a culture dish, air-drying indoors, placing the culture dish on a clean bench for ultraviolet sterilization for 1h to obtain the dry (luteolin) sustained-release algae-inhibiting microspheres.
The luteolin sustained-release algae-inhibiting microsphere provided by the embodiment takes luteolin as a core material and chitosan and sodium alginate as wall materials. The preparation method has simple process and lower cost, and the emulsification step before embedding can ensure that the luteolin is uniformly distributed in the sustained-release algae-inhibiting microspheres, thereby being beneficial to realizing the stable release of the luteolin.
Example 2
The luteolin release characteristics of the sustained-release algal inhibiting microspheres provided in example 1 were tested:
the culture medium is filled with 0.02g/mL sustained-release algae inhibiting microspheres (containing luteolin equivalent 0.41mg/mL), and cultured under illumination of 2000lux (12h light/12 h dark) at 25 +/-1 ℃.
The cumulative amount of luteolin released was measured by changing the same amount of fresh medium every 1.5 days, and the results showed that luteolin released increased to 171.79. mu.g/mL on day 1, then decreased and stabilized at a lower level until day 49, with a slight increase in days 10-13. The accumulated release amount of the microspheres in 22-49 days is 300.27-383.58 mu g/mL, and accounts for 73.32-81.36% of the total content of luteolin in the sustained-release algae-inhibiting microspheres.
The actual concentration of luteolin released can be measured by sampling regularly without changing the culture medium, and the results show that the average concentration of luteolin released in the first 10 days is 168.95 mug/mL, and then the average concentration of luteolin released in the subsequent days generally drops below 100 mug/mL, which indicates that the luteolin released is slowly released and consumed in the culture solution. By calculation, the release time of the luteolin can reach more than 70 days theoretically, and the typical time period of the occurrence of the harmful cyanobacterial bloom can be completely covered.
Example 3
The high-efficiency and lasting inhibition effect of the sustained-release algae-inhibiting microspheres provided in example 1 on the growth of microcystis is tested:
the test algal species: microcystis aeruginosa FACHB-915.
The method comprises the following steps: inoculating Microcystis in exponential growth phase in BG11 culture medium, wherein the initial density of algae cells is 4 × 105The cell/mL meets the algae density of microcystis bloom, the concentration of the sustained-release algae inhibiting microspheres is 0.004g/mL according to the luteolin dose and the release characteristic of the sustained-release algae inhibiting microspheres, meanwhile, treatment groups of the microcystis culture solution containing 0.082mg/mL pure luteolin (equal to the luteolin contained in the sustained-release algae inhibiting microspheres) and 0.004g/mL blank microspheres (not containing the luteolin) and the like are respectively arranged, the microcystis culture solution without any additive is used as a control group, and the microcystis growth comparison experiment of 49 days of the behavior period is carried out under the conditions of 25 +/-1 ℃ and 2000lux (12h light/12 h dark) illumination intensityAnd testing the algae inhibiting effect of the slow-release algae inhibiting microsphere. The dynamic change in cell density of microcystis was measured by visual counting with a microscope (BX53F Olympus, Japan) using a hemocytometer. The Growth Inhibition Ratio (GIR) is calculated by the following formula:
GIR＝(Mt-Nt)/Mt×100％
wherein M istAnd NtThe cell density of the microcystis is of the control group and the treatment group on the t day respectively.
As a result: the results of the cell density of algae under different treatments as a function of the number of days are shown in Table 1. Long-term experiments show that the slow-release algae inhibiting microspheres have a remarkable inhibiting effect on the growth of microcystis from the beginning of the experiment and reach extremely high GIR (98.00-99.92%) from 5 days to 49 days. Therefore, the slow-release algae inhibiting microspheres show extremely strong inhibition effect on the growth of microcystis until the 49 th day.
In contrast, microcystis biomass increased in both the same amount of pure luteolin and blank microsphere treatments, and when exposed to the same amount of pure luteolin, the GIR was only 27.95% -64.94% at days 18-39, after which the inhibition dropped below 25% until the end of the experiment. From day 3 to the end of the experiment, the GIR of the sustained-release algal-inhibiting microspheres was significantly higher than that of pure luteolin contained therein in an equivalent amount (table 2). Compared with the control group, the blank microspheres have no obvious algae killing effect until the experiment is finished. The above proves that only luteolin in the sustained-release algae-inhibiting microspheres plays an algae-inhibiting role, the sustained release of luteolin can greatly enhance and prolong the algae-killing effect of luteolin, the prepared sustained-release algae-inhibiting microspheres have a very obvious algae-inhibiting effect on the growth of blue-green algae, and can realize a long-term lasting (nearly 50 days), extremely high maintenance (GIR > 98%) and stable algae-inhibiting growth effect.
The slow-release algae inhibiting microsphere constructed by the invention can realize the highest and most durable algae inhibiting growth effect of GIR so far.
TABLE 1 algal cell density as a function of days for different treatments
Figure BDA0002939315160000091
TABLE 2 algal cell Growth Inhibition Rate (GIR) with different treatments as a function of days
Figure BDA0002939315160000092
Example 4
The long-term effect of the sustained-release algal inhibiting microspheres provided in example 1 on the release of MCs was tested:
the test algal species: microcystis aeruginosa FACHB-915.
The method comprises the following steps: inoculating Microcystis in exponential growth phase in BG11 culture medium, wherein the initial density of algae cells is 4 × 105The individual cells/mL meet the algae density when the microcystis is in water bloom, according to the luteolin dosage and the release characteristics of the sustained-release algae-inhibiting microspheres, the sustained-release algae-inhibiting microspheres are added into the microcystis culture solution to enable the concentration of the microcapsule algae culture solution to reach 0.004g/mL, meanwhile, a microcystis culture solution treatment group containing 0.082mg/mL pure luteolin (equal to the luteolin contained in the sustained-release algae-inhibiting microspheres) and the microcystis culture solution without any additive are arranged as a control group, and a comparison experiment of releasing MCs by the algae cells in the behavior period of 49 days is carried out under the conditions of 25 +/-1 ℃ and illumination intensity of 2000lux (12h light/12 h dark), so that the influence of the sustained-release algae-inhibiting microspheres on the release of the MCs by the algae cells is tested. Algal cell culture fluid was collected before (day 15), during (day 23), after (day 39), and at the end (day 49) of the experiment, centrifuged at 8000rpm for 10min to retain supernatant and collect algal cells, and the supernatant was directly used for the extracellular MCs assay to characterize the release of MCs. And (3) carrying out intracellular and extracellular MCs concentration determination by using enzyme-linked immunosorbent assay.
As a result: the results of the change in extracellular MCs concentration with the number of days for the different treatments are shown in Table 3. Long-term experiments show that compared with the equivalent pure luteolin, the slow-release algae-inhibiting microspheres have higher inhibition effect on the release of microcystis MCs in the front, middle, back and final stages, the inhibition rate of the slow-release algae-inhibiting microspheres on the release of MCs is up to 98.88-99.55%, and the slow-release algae-inhibiting microspheres continue to the 49 th day (table 4). The concentration of extracellular MCs in the front, middle, back and end stages is only 0.84-1.30 mug/L (table 3), the range is close to or lower than the safety threshold (1 mug/L) of the concentration of MCs in drinking water recommended by the world health organization, and the ecological safety significance is great.
TABLE 3 extracellular MCs concentration as a function of days for different treatments
Figure BDA0002939315160000101
TABLE 4 inhibition of MCs Release by algal cells under different treatments as a function of days
Figure BDA0002939315160000102
The release of MCs from algae cells to water bodies can cause huge ecological risks, and long-term experimental results prove that the slow-release algae inhibiting microspheres can efficiently, durably and stably inhibit microcystis from releasing MCs, the release inhibition rate is up to more than 98.5 percent, the duration is up to 49 days, the highest and most durable MCs release inhibition effect is achieved so far, and the slow-release algae inhibiting microspheres are extremely effective in repairing the pollution of the MCs in the water bodies.
Example 5
The high-efficiency and lasting inhibition effect of the slow-release algae inhibiting microspheres provided in example 1 on the growth of microcystis and the release of MCs under different nitrogen levels is tested:
the test algal species: microcystis aeruginosa FACHB-915.
The method comprises the following steps: inoculating Microcystis in exponential growth phase into BG11 culture medium with nitrogen concentration of 0.5mg/L, 5mg/L and 50mg/L respectively at initial density of 4 × 105One cell/mL, corresponding to the algal density of the microcystis bloom. According to the dose and the release characteristics of luteolin in the sustained-release algae-inhibiting microspheres, the sustained-release algae-inhibiting microspheres are respectively added into the micro-capsule algae culture solution with each nitrogen concentration to enable the concentration to reach 0.003g/mL, meanwhile, a micro-capsule algae culture solution treatment group containing 0.061mg/mL pure luteolin (equal to the luteolin contained in the sustained-release algae-inhibiting microspheres) and the micro-capsule algae culture solution without any additive are arranged for each nitrogen concentration to serve as a control group, a comparison experiment of algae cell growth in the behavior period of 30 days is carried out under the conditions of 25 +/-1 ℃ and 2000lux (12h light/12 h dark) illumination intensity, and the test of different nitrogen levels is carried outThe influence of the lower sustained-release algae inhibiting microspheres on the growth of algae cells. The dynamic change in cell density of microcystis was measured by visual counting with a microscope (BX53F Olympus, Japan) using a hemocytometer. The Growth Inhibition Rate (GIR) was calculated by the following formula:
GIR＝(Mt-Nt)/Mt×100％
wherein M istAnd NtThe cell density of the microcystis is of the control group and the treatment group on the t day respectively. Algal cell culture fluid was collected at the middle (day 20) and end (day 30) of the experiment, centrifuged at 8000rpm for 10min to retain supernatant and collect algal cells, and the supernatant was used directly for extracellular MCs assay to characterize release of MCs. And (3) carrying out intracellular and extracellular MCs concentration determination by using enzyme-linked immunosorbent assay.
As a result: the results of the cell density of algae treated differently at different nitrogen levels as a function of the number of days are shown in tables 5, 6, 7. Long-term experiments show that the slow-release algae-inhibiting microspheres under different nitrogen levels can obviously inhibit the growth of microcystis from the beginning of the experiment, and reach extremely high GIR (more than 90%) from day 8 to day 30, and have extremely high inhibition on the growth of microcystis until day 30, and the extremely high GIR is not influenced by the change of the nitrogen level and the days (Table 8). In addition, compared with a control group, the average inhibition rate of CRM on MCs release at 20 th to 30 th days under different nitrogen levels is also extremely high, and the average inhibition rate is more than 90%, and the influence of nitrogen level change and days is not obvious.
In contrast, the microcystis biomass showed a significant increase in the same amount of pure luteolin treatment at different nitrogen levels, and the GIRs were only 33.78% -72.60%, 42.34% -76.30% and 41.06% -78.51% at 8-30 days at nitrogen levels of 0.5, 5 and 50mg/L, respectively (Table 8), and were less than 60% after 16 days, when exposed to the same amount of pure luteolin. From day 8 to the end of the experiment, the GIR of the sustained-release algal-inhibiting microspheres was significantly higher than that of pure luteolin contained therein in an equivalent amount (table 8). In addition, the inhibition rate of the equivalent pure luteolin on the release of MCs at 20-30 days under different nitrogen levels is obviously lower than that of CRM.
TABLE 5 Nitrogen concentration of 50mg/L, algal cell density with different treatments as a function of days
Figure BDA0002939315160000121
TABLE 6 Nitrogen concentration of 5mg/L, algal cell density with different treatments as a function of days
Figure BDA0002939315160000122
TABLE 7 Nitrogen concentration of 0.5mg/L, algal cell density with different treatments as a function of days
Figure BDA0002939315160000123
Figure BDA0002939315160000131
The microcystis bloom is often burst in water bodies with different nitrogen levels (different eutrophication degrees), and the finding of stable, efficient and lasting algae inhibiting effect suitable for different nitrogen levels is significant. The long-term experiment result shows that the prepared slow-release algae inhibiting microspheres have very obvious inhibiting effects on the growth of blue-green algae and the release of MCs under different nitrogen levels, maintain extremely high GIR (more than 90 percent) and have the duration of at least 30 days.
The slow-release algae inhibiting microsphere constructed by the invention can realize highest and most lasting algae inhibiting growth and toxin inhibiting release effects of GIR under different nitrogen levels so far, and has remarkable positive effects on preventing and treating microcystis bloom in water bodies with different nutrition degrees.
TABLE 8 algal cell Growth Inhibition (GIR) at different nitrogen levels and different treatments as a function of days
Figure BDA0002939315160000132
The slow-release algae inhibiting microsphere provided by the invention can greatly prolong and improve the action time and effect of luteolin under different environmental conditions, and realizes long-term effective algae inhibition and MCs release inhibition.
Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.

Claims (10)

1. The slow-release algae-inhibiting microsphere is characterized by being prepared from the following raw materials:
luteolin 1% ~1.5% (w/v)
Olive oil 18% ~20% (w/v)
TWEEN-80 10%~12% (w/v)
Methanol 10% ~12% (w/v)
Sodium alginate 1% -2.5% (w/v)
0.2% -0.3% (w/v) of chitosan
Calcium chloride 4% ~8% (w/v)
0.5% -1% (v/v) of glutaraldehyde.
2. The preparation method of the sustained-release algal-inhibiting microsphere of claim 1, comprising:
1) dissolving luteolin in olive oil, sequentially adding methanol and TWEEN-80, mixing uniformly and ultrasonically treating to form a uniform luteolin emulsifying system, adding the luteolin emulsifying system into a sodium alginate solution, and stirring while fully mixing to obtain a mixed solution;
2) and dropwise adding the mixed solution into an aqueous solution of chitosan and calcium chloride to form microspheres, uniformly shaking while dropwise adding, adding glutaraldehyde, stirring, and performing aftertreatment.
3. The preparation method of the sustained-release algal-inhibiting microsphere according to claim 2, wherein in the step 1), the ultrasonic frequency is 150W in the ultrasonic treatment; the stirring speed is 100-200 r/min, and the stirring time is 0.5-1 min.
4. The preparation method of the sustained-release algal-inhibiting microsphere according to claim 3, wherein in the step 1), the preparation of the sodium alginate solution comprises the following steps: mixing sodium alginate powder with water, and heating with outer flame of alcohol lamp until sodium alginate and water are completely mixed and dissolved.
5. The preparation method of the slow-release algae-inhibiting microsphere according to claim 2, wherein in the step 2), the stirring speed is 100-200 r/min; and slowly dripping the mixed solution into an aqueous solution containing chitosan and calcium chloride at a position 10-20 cm higher than the liquid level.
6. The method for preparing the sustained-release algal-inhibiting microspheres according to claim 5, wherein in the step 2), the mixed solution is slowly dropped into an aqueous solution containing chitosan and calcium chloride at a position 15cm higher than the liquid level.
7. The preparation method of the sustained-release algal-inhibiting microsphere according to claim 5, wherein the post-treatment comprises: and washing, filtering, drying and sterilizing the microspheres.
8. The preparation method of the slow-release algae-inhibiting microsphere according to claim 7, wherein the microsphere is obtained by washing the microsphere with petroleum ether for 2-5 times, washing the microsphere with distilled water for 2-5 times, then carrying out vacuum filtration, carrying out air drying at room temperature, and carrying out ultraviolet sterilization.
9. The preparation method of the sustained-release algal-inhibiting microsphere according to claim 8, wherein the microsphere is prepared by washing the microsphere with petroleum ether for 3 times, washing the microsphere with distilled water for 3 times, then performing vacuum filtration, air-drying at room temperature, and performing ultraviolet sterilization.
10. The use of the sustained-release algal-inhibiting microspheres according to claim 1 or the sustained-release algal-inhibiting microspheres obtained by the preparation method of the sustained-release algal-inhibiting microspheres according to any one of claims 2 to 9 in long-acting prevention and control of algal bloom and inhibition of toxin release thereof.
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