CN113564052B - Microalgae directional culture solution and application thereof, device for preparing sludge hydrolysate and device for directionally culturing and enriching biomass microalgae - Google Patents

Microalgae directional culture solution and application thereof, device for preparing sludge hydrolysate and device for directionally culturing and enriching biomass microalgae Download PDF

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CN113564052B
CN113564052B CN202110889988.1A CN202110889988A CN113564052B CN 113564052 B CN113564052 B CN 113564052B CN 202110889988 A CN202110889988 A CN 202110889988A CN 113564052 B CN113564052 B CN 113564052B
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microalgae
hydrolysate
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cytokinin
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CN113564052A (en
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陈秀荣
胡雪洋
田金乙
张心雨
马丽娟
顾昊
杨莹莹
唐宇琛
潘涛
丁彪
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East China University of Science and Technology
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Abstract

The invention relates to the technical field of microalgae culture, in particular to a microalgae directional culture solution and application thereof, a device for preparing sludge hydrolysate and a device for directionally culturing and enriching biomass microalgae. The invention provides a microalgae directional culture solution, which comprises a sludge hydrolysate and a microelement supplementing solution containing cytokinin; the volume ratio of the sludge hydrolysate to the microelement supplementing liquid containing cytokinin is (1-1.6): 1. The culture solution provided by the invention can obviously improve the yield of microalgae value-added biomass by utilizing the sludge hydrolysate, cytokinin and trace element supplementing liquid, and can utilize a large amount of sludge by utilizing the sludge hydrolysate, and the cost is relatively low, so that the sludge is utilized as a resource.

Description

Microalgae directional culture solution and application thereof, device for preparing sludge hydrolysate and device for directionally culturing and enriching biomass microalgae
Technical Field
The invention relates to the technical field of microalgae culture, in particular to a microalgae directional culture solution and application thereof, a device for preparing sludge hydrolysate and a device for directionally culturing and enriching biomass microalgae.
Background
Along with the development of China, the discharge amount of the industrial waste mud of the sewage plant is also improved. Although the method for treating wastewater by using microorganisms can reduce the concentration of nitrogen and phosphorus in wastewater, the microorganism nitrogen-containing secondary metabolites and phosphorus still accumulate in the residual sludge, a large amount of residual sludge is difficult to treat, the treatment cost is high, and the residual toxicity is high.
The microalgae has the characteristics of high photosynthesis efficiency, short growth period, no need of occupying agricultural land, strong environment adaptability and the like. Microalgae is easy to optimize, regulate and control, improves yield, is easy to crush and dry, and can produce high-value byproducts such as proteins, pigments and the like. Among all biodiesel feedstocks, microalgae have high grease productivity and fix carbon dioxide during growth and accumulation of grease.
How to increase the yield of microalgae-based value-added biomass is a major problem faced by microalgae cultivation. If the sludge can be utilized to culture microalgae, not only the nutrient elements such as carbon, nitrogen, phosphorus and the like in the sludge can be recycled, but also the yield of the microalgae value-added biomass can be improved, so that the purpose of industrial operation of the microalgae can be achieved. Therefore, a need for a microalgae culture method that can utilize sludge to increase microalgae value-added biomass is currently felt.
Disclosure of Invention
In order to solve the problems, the invention provides a microalgae directional culture solution, application thereof, a device for preparing sludge hydrolysate and a device for directionally culturing and enriching biomass microalgae. The culture solution can remarkably improve the yield of microalgae value-added biomass by utilizing the sludge hydrolysate and the microelement supplement solution containing cytokinin.
In order to achieve the above object, the present invention provides the following technical solutions:
The invention provides a microalgae directional culture solution, which comprises a sludge hydrolysate and a microelement supplementing solution containing cytokinin; the trace element supplementing liquid comprises the following components in mass: 120-150 g/L NaNO 3, 3-5 g/L K 2HPO4, 5-8 g/L MgSO 4·7H2 O, 2-5 g/L CaCl 2·2H2 O, 0.4-0.7 g/L citric acid, 0.5-0.8 g/L ferric ammonium citrate, 0.1-0.2 g/L EDTANa 2, 1-3 g/L Na 2CO3;
the mass percentage of cytokinin in the microelement supplementing liquid containing cytokinin is 2% -5% of the mass of the microelement supplementing liquid;
the volume ratio of the sludge hydrolysate to the microelement supplementing liquid containing cytokinin is (1-1.6): 1.
Preferably, the cytokinin comprises one or more of kinetin, zeatin and 6-aminobenzyl purine.
Preferably, the preparation of the sludge hydrolysate comprises the following steps:
Standing the sludge to obtain concentrated sludge;
Sequentially adjusting the pH value, carrying out ultrasonic crushing and anaerobic hydrolysis on the concentrated sludge to obtain sludge hydrolysate; the pH value of the primary sludge obtained after the adjustment is more than or equal to 7.
Preferably, the ultrasound is intermittent ultrasound; the intermittent ultrasound is ultrasound for 4s, and pausing for 4s; in the ultrasonic treatment, the ultrasonic frequency is 10-30 kHz, the power is 50-80W, the time is preferably 3-8 min, and the temperature of the primary sludge obtained after the pH value is regulated is less than 50 ℃.
Preferably, the ultrasonic crushing further comprises adding H 2O2 into the intermediate sludge obtained after the ultrasonic crushing; the addition amount of the H 2O2 is 0.1-0.3 mL/gVSS.
The invention provides application of the microalgae directional culture solution in directional culture of biomass-enriched microalgae, wherein the microalgae comprise green algae.
The invention provides a method for directionally culturing and enriching biomass microalgae, which comprises the following steps:
And inoculating microalgae into the culture solution, and culturing.
The invention provides an application of microalgae cultured by the method in biofertilizer.
The invention provides a device for preparing sludge hydrolysate, which comprises a sludge ultrasonic crushing unit 1 and a sludge anaerobic hydrolysis unit 2 which are communicated; the sludge ultrasonic crushing unit 1 comprises a primary sludge storage device 3 and a first stirrer 4; the sludge anaerobic hydrolysis unit 2 comprises a sludge anaerobic hydrolyzer 5, an H 2O2 feed inlet 6 and a second stirrer 7.
The invention provides a device for directionally culturing and enriching biomass microalgae, which comprises a sludge hydrolysate liquid storage unit 9, a microelement supplementing liquid storage unit 10 and a microalgae culturing unit 11; the sludge hydrolysate liquid storage unit 9 and the trace element supplementing liquid storage unit 10 are respectively communicated with the microalgae culture unit 11;
the sludge hydrolysate storage unit 9 comprises a sludge hydrolysate reservoir 12; the trace element supplementing liquid storage unit 10 comprises a trace element supplementing liquid storage 15; the microalgae culturing unit 11 comprises a feed inlet 18, a flash illumination incubator 19, a microalgae inoculation inlet 21 and a discharge outlet 22; the flash light incubator 19 includes a variable frequency light source 20.
The beneficial effects are that: the invention provides a microalgae directional culture solution, which comprises a sludge hydrolysate and a microelement supplementing solution containing cytokinin; the trace element supplementing liquid comprises the following components in mass: 120-150 g/L NaNO 3, 3-5 g/L K 2HPO4, 5-8 g/L MgSO 4·7H2 O, 2-5 g/L CaCl 2·2H2 O, 0.4-0.7 g/L citric acid, 0.5-0.8 g/L ferric ammonium citrate, 0.1-0.2 g/L EDTANa 2, 1-3 g/L Na 2CO3; the mass percentage of cytokinin in the microelement supplementing liquid containing cytokinin is 2% -5% of the mass of the microelement supplementing liquid; the volume ratio of the sludge hydrolysate to the microelement supplementing liquid containing cytokinin is (1-1.6): 1. According to the invention, the algae and bacteria in the sludge hydrolysate form a algae symbiotic system to remove the sludge toxicity, so that the sludge biotoxicity is reduced from 80% to about 10%; the components in the microelement supplementing liquid containing cytokinin can meet the growth requirement of microalgae in anaerobic hydrolysate, and provide microelement for the growth of microalgae; cytokinin is a plant hormone, can promote green algae to synthesize biomass such as glutamic acid, starch, pigment and the like, and can supplement microelement supplementing liquid containing cytokinin to promote the growth of green algae and obtain value-added biomass such as protein, polysaccharide, carotenoid and the like; therefore, the culture solution can remarkably improve the yield of microalgae value-added biomass by utilizing the sludge hydrolysate, cytokinin and trace element supplementing liquid, and a large amount of surplus sludge can be utilized by utilizing the sludge hydrolysate, so that the cost is relatively low, and the surplus sludge is recycled.
In addition, the invention provides a preparation method of the sludge hydrolysate, which can better break the sludge and release the nutrient substances contained in the sludge.
The invention also provides a microalgae culture method, which can save microalgae culture cost, capture biomass value-added products of glutamic acid proteins up to 400mg/L, starch 200mg/L and carotenoid 5mg/L, realize ecological recycling and reduction utilization of residual sludge, and capture microalgae biomass as a biofertilizer which can be applied to agricultural production in farmlands, promote crop growth and increase yield.
Drawings
Fig. 1 is a diagram of a device for preparing a sludge hydrolysate, wherein 1 is a sludge ultrasonic crushing unit, 2 is a sludge anaerobic hydrolysis unit, 3 is a primary sludge storage device, 4 is a first stirrer, 5 is a sludge anaerobic hydrolysis device, 6 is an H 2O2 feed inlet, 7 is a second stirrer, and 8 is a sludge inlet;
fig. 2 is a diagram of a device for microalgae cultivation, 9 is a sludge hydrolysate liquid storage unit, 10 is a microelement supplementing liquid storage unit, 11 is a microalgae cultivation unit, 12 is a sludge hydrolysate liquid storage device, 13 is a sludge hydrolysate discharge port, 14 is a flowmeter, 15 is a microelement supplementing liquid storage device, 16 is a microelement supplementing liquid discharge port, 17 is a diaphragm pump, 18 is a feed port, 19 is a flash light illumination incubator, 20 is a variable frequency light source, 21 is a microalgae inoculation port, and 22 is a discharge port.
Detailed Description
The components, medicaments and devices of the invention are obtained by routine purchase by a person skilled in the art, unless otherwise specified.
The invention provides a microalgae directional culture solution, which comprises the sludge hydrolysate and a microelement supplementing solution containing cytokinin; the trace element supplementing liquid comprises the following components in mass: 120-150 g/L NaNO 3, 3-5 g/L K 2HPO4, 5-8 g/L MgSO 4·7H2 O, 2-5 g/L CaCl 2·2H2 O, 0.4-0.7 g/L citric acid, 0.5-0.8 g/L ferric ammonium citrate, 0.1-0.2 g/L EDTANa 2, 1-3 g/L Na 2CO3, preferably comprising the following components by mass: 125-145 g/L NaNO 3, 3.5-4.5 g/L K 2HPO4, 5.5-7.5 g/L MgSO 4·7H2 O, 2.5-4.5 g/L CaCl 2·2H2 O, 0.45-0.65 g/L citric acid, 0.55-0.75 g/L ferric ammonium citrate, 0.12-0.18 g/L EDTANa 2, 1.5-2.5 g/L Na 2CO3, more preferably comprising the following components by mass: 130-140 g/L NaNO 3, 3.8-4.2 g/L K 2HPO4, 6-7 g/L MgSO 4·7H2 O, 3-4 g/L CaCl 2·2H2 O, 0.5-0.6 g/L citric acid, 0.6-0.7 g/L ferric ammonium citrate, 0.14-0.16 g/L EDTANa 2, 1.9-2.2 g/L Na 2CO3.
In the invention, the mass percentage of cytokinin in the microelement supplementing liquid containing cytokinin is 2% -5% of the mass of the microelement supplementing liquid, preferably 2.5% -4.5%, more preferably 3% -4%; the volume ratio of the sludge hydrolysate to the microelement supplementing liquid containing cytokinin is (1-1.6): 1, preferably (1.2-1.5): 1; the cytokinin preferably comprises one or more of kinetin, zeatin and 6-aminobenzyl purine.
The growth of microalgae needs a plurality of microelements, and although the sludge hydrolysate contains rich nutrient elements such as carbon, nitrogen, phosphorus and the like, the growth requirement of the microalgae in the anaerobic hydrolysate cannot be fully met, and the microelements are provided for the growth of the microalgae by supplementing NaNO 3、K2HPO4、MgSO4、CaCl2·2H2 O and the like; cytokinin is a plant hormone, and the cytokinin is supplemented to promote green algae to synthesize biomass such as glutamic acid, starch, pigment and the like, improve the capability of microalgae for resisting plant stress, promote the activity of glutamate dehydrogenase, further synthesize a large amount of glutamic acid, and promote the generation of starch and carotenoid, so that the growth of green algae can be promoted and the value-added biomass such as protein, polysaccharide, carotenoid and the like can be obtained by adding trace element supplementing liquid. Therefore, the sludge hydrolysate and the culture solution can be used for directional culture and enrichment biomass culture of microalgae, and industrial surplus sludge hydrolysate culture of microalgae enrichment value-added biomass is an environment-friendly biomass and energy enrichment way.
The invention has no special requirement on the source of the sludge hydrolysate, and is well known to the person skilled in the art or prepared by self. When the sludge hydrolysate is provided by adopting a self-preparation mode; the preparation method of the sludge hydrolysate preferably comprises the following steps:
Standing the sludge to obtain concentrated sludge;
and sequentially adjusting the pH value, carrying out ultrasonic crushing and anaerobic hydrolysis on the concentrated sludge to obtain sludge hydrolysate.
In the invention, the sludge is preferably subjected to standing treatment to obtain the concentrated sludge. The sludge is preferably surplus sludge of an industrial wastewater treatment plant; the VSS/TSS of the sludge is more than 70%; the sludge is preferably residual sludge after the secondary biochemical treatment of the industrial wastewater after heavy metal pre-removal, and the heavy metal of the sludge reaches the agricultural sludge pollutant control standard (GB 4284-2018). In the present invention, the time of the standing is preferably 4 to 8 hours, more preferably 6 to 8 hours; the temperature of the standing is preferably 25-30 ℃; the water content of the concentrated sludge is more than or equal to 97 wt%. The concentrated sludge is preferably obtained by standing sludge and pouring supernatant.
The invention preferably carries out pH value adjustment, ultrasonic crushing and anaerobic hydrolysis on the concentrated sludge in sequence to obtain sludge hydrolysate. In the invention, the pH value of the primary sludge obtained after the pH value adjustment is preferably more than or equal to 7, more preferably 7-9; the pH is adjusted to make the hydrolysis environment weak base, so that the sludge hydrolysis efficiency is improved. In the present invention, the intermittent ultrasound is preferably ultrasound for 4s, and the intermittent ultrasound is suspended for 4s, the present invention uses ultrasound to destroy cells to release EPS, and low-intensity ultrasound can prevent bacteria from being killed completely. In the ultrasonic treatment, the ultrasonic frequency is preferably 10-30 kHz, more preferably 20kHz; the power is preferably 50 to 80W, more preferably 60W; the time is preferably 3 to 8 minutes, more preferably 5 minutes; the temperature of the concentrated sludge is preferably < 50 ℃, more preferably > 10 ℃ during ultrasound; the temperature of the concentrated sludge during ultrasonic treatment is preferably controlled by an ice water bath. In order to better crack the sludge and release EPS, release nutrient elements such as carbon, nitrogen, phosphorus and the like, the concentrated sludge is subjected to ultrasonic crushing before anaerobic hydrolysis, and meanwhile, in order to avoid the total bacterial cracking and death caused by the overhigh ultrasonic strength, low-intensity ultrasonic is adopted.
The method is characterized by preferably adding H 2O2 into the medium-grade sludge obtained after ultrasonic crushing; the amount of H 2O2 added is preferably 0.1 to 0.3mL/g VSS, i.e., 0.1 to 0.3mL H 2O2, more preferably 0.15mL/g VSS, is added to 1g Volatile Suspended Solids (VSS) in the sludge. H 2O2 is added, and the oxidizing property of the H 2O2 is utilized to further crack and release N, P, SCOD and the like of sludge cells.
In the present invention, the temperature of the anaerobic hydrolysis is preferably a medium temperature, more preferably 30 to 45 ℃, and most preferably 35 to 38 ℃; the shaking speed of the anaerobic hydrolysis is preferably > 100rpm, more preferably 150rpm; the time of the anaerobic hydrolysis is preferably 3d; the anaerobic method adopts a nitrogen blowing mode preferably; the nitrogen blowing time is preferably 5min; the method of nitrogen blowing is not particularly limited in the present invention, and the method is well known to those skilled in the art.
The anaerobic hydrolysis preferably further comprises centrifuging the intermediate sludge obtained by the anaerobic hydrolysis; the rotational speed of the centrifugation is preferably > 5000rpm, more preferably 8000-12000 rpm; the time for the centrifugation is preferably 5 to 15 minutes, more preferably 6 to 14 minutes, and still more preferably 7 to 13 minutes. According to the invention, after anaerobic hydrolysis, centrifugation is carried out, so that elements such as carbon, nitrogen, phosphorus and the like released by the sludge can be better solubilized into the sludge hydrolysate.
According to the invention, the sludge hydrolysate containing nutrient substances such as nitrogen, phosphorus, potassium and the like can be obtained in an anaerobic hydrolysis mode, toxic substances accumulated in the sludge are released, and macromolecular substances such as polysaccharide, protein and the like which form biotoxicity in the sludge can be converted into micromolecular substances under an anaerobic condition so as to be absorbed and utilized by microalgae, thereby synthesizing biomasses such as protein, starch and the like. The traditional treatment (incineration and landfill) of the excess sludge has high cost, and the cost of generating hydrolysate by utilizing anaerobic hydrolysis of the sludge is relatively low, so that the sludge can be recycled.
The sludge hydrolysate prepared by the invention preferably comprises the following components in mass: 359.7 to 368.7Mg/L COD, 141.4 to 147.6Mg/L soluble protein, 72 to 75.8Mg/L soluble polysaccharide, 69.4 to 83.6Mg/L NH 3 -N, 262.5 to 280.5Mg/L TN, 9.8 to 10.2Mg/L PO 4 -P, 13.2 to 18.2Mg/L TP, 36 to 46Mg/L Ca, 69 to 75Mg/L Mg, 22 to 34Mg/L Fe, 4 to 5.2Mg/L Mn, more preferably comprising the following components by mass: 362.5-366.5 Mg/L COD, 143.5-146.8 Mg/L soluble protein, 72.5-74.8 Mg/L soluble polysaccharide, 74.3-82.5 Mg/L NH 3 -N, 268.6-276.5 Mg/L TN, 9.8-10.2 Mg/L PO 4 -P, 13.4-14.2 Mg/L TP, 39-43 Mg/L Ca, 72-75 Mg/L Mg, 30-34 Mg/L Fe, 4.4-5.2 Mg/L Mn; the toxicity of the sludge hydrolysate is preferably 82.9+/-0.9%. Organic matters such as COD, protein, polysaccharide and the like in the sludge hydrolysate can be absorbed and utilized by microalgae as nutrient substances, NH 3-N、PO4 -P, TN and TP are taken as microalgae survival nutrient elements, ca, mg, fe, mn is taken as trace elements and can be absorbed and utilized by microalgae, so that the growth requirement of the microalgae is met, and the microalgae is beneficial to synthesizing organic matters; the sludge hydrolysate contains rich nutrient elements such as carbon, nitrogen, phosphorus and the like, and various micromolecular substances such as amino acid, acetic acid, propionic acid and the like which are obtained by hydrolyzing toxic secondary metabolites such as antibiotics, stress proteins, SMP and the like secreted by microorganisms when the microorganisms are stressed or impacted can be well utilized by microalgae, so that biomass such as protein, polysaccharide, carotenoid and the like in microalgae cells is increased.
In the invention, the preparation of the sludge hydrolysate is preferably carried out in a device for preparing the sludge hydrolysate, and the device is simple and convenient for sludge treatment. As a preferred embodiment of the present invention, the apparatus for preparing a sludge hydrolysate is preferably comprised of a sludge ultrasonic crushing unit 1 and a sludge anaerobic hydrolysis unit 2 in communication, as shown in FIG. 1.
In the present invention, the mud ultrasonic crushing unit 1 preferably includes a primary sludge storage tank 3 and a first agitator 4; the primary sludge storage device 3 is preferably a primary sludge storage box; the first stirrer 4 is preferably an ultrasonic breaker horn, and the ultrasonic breaker horn is preferably an ultrasonic breaker; the ultrasonic breaker is preferably purchased from Shanghai analysis ultrasonic instruments, inc. of China, model number is preferably FS-300N. In the invention, concentrated sludge is preferably put into a primary sludge storage box, and is subjected to ultrasonic crushing by using an amplitude transformer of an ultrasonic crusher.
In the present invention, the sludge anaerobic hydrolysis unit 2 preferably includes a sludge anaerobic hydrolyzer 5, an H 2O2 feed inlet 6, and a second agitator 7; the sludge anaerobic hydrolyzer 5 preferably comprises a sludge anaerobic hydrolysis tank; the sludge ultrasonic crushing unit 1 is further preferably communicated with a sludge anaerobic hydrolyzer through a sludge inlet 8. In the invention, the medium-grade sludge subjected to ultrasonic crushing is preferably conveyed to a sludge anaerobic hydrolysis tank through a sludge inlet 8; the medium-grade sludge to be conveyed and ultrasonically crushed enters a sludge anaerobic hydrolysis tank, is introduced into H 2O2 through an H 2O2 feed inlet 6, and is subjected to anaerobic hydrolysis under the stirring action of a second stirrer 7.
The preparation method of the sludge hydrolysate can better break the sludge and release the nutrient substances contained in the sludge.
The invention provides an application of the culture solution in directional culture of biomass-enriched microalgae, wherein the microalgae comprise green algae, preferably one or more of Chlorella (Chlorella), scenedesmus (Scenedesmus), green algae (Chlorococcum), chlamydomonas (Chlamydomonas) and Dunaliella (Dunaliella); the chlorella preferably comprises chlorella pyrenoidosa; the scenedesmus preferably comprises scenedesmus obliquus; the Chlorella preferably comprises Chlorella terrestris. The source of the microalgae is not particularly limited, and the microalgae can be obtained by routine purchase by a person skilled in the art; in a specific embodiment of the invention the microalgae are preferably purchased from the Proc of the Chinese institute of aquatic life fresh water algae seed stock (FACHB).
The invention provides a method for directionally culturing and enriching biomass microalgae, which comprises the following steps:
And inoculating microalgae into the culture solution, and culturing.
In the inoculation process, the density of the microalgae is preferably more than 1 multiplied by 10 6 cells/mL, more preferably (2-3) multiplied by 10 6 cells/mL; the dry weight of the microalgae is preferably greater than about 0.03g/L, more preferably about 0.05 to about 0.08g/L. The invention can construct the symbiotic complex of the algae without sterilizing the culture solution during inoculation. The invention utilizes the bacteria in microalgae and hydrolysate to form a bacteria-algae symbiotic system to remove the sludge toxicity, can degrade organic substances, and remove the sludge toxicity, and the relative light inhibition rate of luminescent bacteria is used for characterizing the biological toxicity of the sludge, so that the biological toxicity of the sludge is reduced from 80% to about 10%.
In the present invention, the temperature of the culture is preferably > 20 ℃, more preferably 24 to 26 ℃; the time of the culture is preferably > 5d, more preferably 15 to 20d; the rotation speed of the culture is preferably 120 to 150rpm, more preferably 125 to 145rpm.
In the culture of the invention, the flash frequency range is preferably 30-200 Hz, more preferably 50-100 Hz; the light intensity is preferably > 2000Lux, more preferably 3500-4500 Lux, and the light-dark ratio is preferably (0.8-1.5): 1 (i.e. light time: dark time is preferably (8-15) h:10 h), more preferably 1.4:1 (i.e. light time: dark time is more preferably 14h:10 h). The advantage of the method for culturing microalgae is that low-frequency flashing can induce green algae to generate more biomass such as protein, polysaccharide, carotenoid and the like. The incubator used for the culture is not particularly limited, and can be purchased conventionally by a person skilled in the art; in the specific embodiment of the invention, a flash illumination incubator is preferably adopted in the cultivation; the flash light incubator is preferably available from Sibirk corporation, model number SPX-250B-G.
In the present invention, the directional culture of the biomass-enriched microalgae is preferably performed in a device for directional culture of biomass-enriched microalgae. The device for directionally culturing and enriching the biomass microalgae can uniformly mix the sludge hydrolysate and the microelement supplement liquid containing cytokinin, is convenient to inoculate, can provide proper growth conditions for the microalgae, and can induce the green algae to generate more biomass such as protein, polysaccharide, carotenoid and the like by low-frequency flashing of the incubator. As a preferred embodiment of the invention, the device for directionally culturing and enriching biomass microalgae is shown in fig. 2, and comprises a sludge hydrolysate liquid storage unit 9, a microelement supplementing liquid storage unit 10 and a microalgae culturing unit 11; the sludge hydrolysate liquid storage unit 9 and the microelement supplementing liquid storage unit 10 are respectively communicated with the microalgae culture unit 11.
In the invention, the sludge hydrolysate liquid storage unit 9, the trace element supplementing liquid storage unit 10 and the microalgae culture unit 11 are preferably communicated through pipelines. In the present invention, the sludge hydrolysate storage unit 9 includes a sludge hydrolysate reservoir 12; the sludge hydrolysate reservoir 12 is preferably a sludge hydrolysate reservoir; the sludge hydrolysate liquid storage unit 9 is preferably connected with the microalgae culture unit 11 through a diaphragm pump 17 after flowing out from a sludge hydrolysate discharge port 13; the sludge hydrolysate liquid storage unit 9 is further preferably connected with the microalgae culture unit 11 through a flow meter 14 and a diaphragm pump 17 by flowing out of a sludge hydrolysate discharge hole 13.
In the present invention, the trace element replenishment liquid reservoir unit 10 includes a trace element replenishment liquid reservoir 15; the trace element supplementing liquid reservoir 15 is preferably a trace element supplementing liquid reservoir; the microelement supplementing liquid storage unit 10 preferably flows out through a microelement supplementing liquid discharge port 16 and is connected with the microalgae culture unit 11 through a diaphragm pump 17; the trace element supplementing liquid storage unit 10 is further preferably connected with the microalgae culturing unit 11 through a flow meter 14 and a diaphragm pump 17 by flowing out of a trace element supplementing liquid discharge hole 16. In the invention, a sludge hydrolysate liquid storage unit 9 and a microelement supplement liquid storage unit 10 are respectively connected with a microalgae culture unit 11 through a flowmeter 14 and a diaphragm pump 17, the flow rate of the sludge hydrolysate liquid storage unit and the microelement supplement liquid storage unit can be controlled so as to control the ratio of the microalgae culture unit and the microelement supplement liquid storage unit, and the sludge hydrolysate liquid storage unit and the microelement supplement liquid storage unit are conveyed into a flash illumination incubator 19 from a feed inlet 18 under the action of the diaphragm pump 17 and are mixed to obtain the microalgae directional culture liquid.
In the present invention, the microalgae culturing unit 11 comprises a feed inlet 18, a flash light incubator 19, a microalgae inoculation port 21 and a discharge port 22; the flash light incubator 19 is preferably a flash light incubator; the flash illumination incubator 19 preferably includes a variable frequency light source 20; the variable frequency light source 20 is preferably a flash light emitting diode. In the present invention, the feed port 18 is preferably located at the upper part of the flash incubator 19, and allows the sludge hydrolysate and the trace element supplement liquid to enter the incubator; the microalgae inoculation opening 21 is preferably positioned at the top of the flash illumination incubator, so that microalgae can be conveniently inoculated, and meanwhile, the algae liquid can be ensured to enter the incubator culture liquid from the inoculation opening; the discharge port 22 is preferably positioned at the lower part of the flash light incubator, and can collect the cultured algae liquid. The invention reasonably sets the positions of the feed inlet, the discharge outlet and the inoculation opening, so that microalgae can fully contact with the culture solution, and further effective substances in the culture solution are utilized. The microalgae is preferably connected into the microalgae directional culture solution through a microalgae inoculation opening 21 at the left side of the top of the flash illumination incubator 19, and the cultured microalgae biomass is obtained through a discharge opening 22 at the right side of the flash illumination incubator 19.
In the present invention, the flash light incubator 19 may be plural; when a plurality of flash illumination incubators exist, each flash illumination incubator 19 is respectively communicated with the sludge hydrolysate liquid storage unit 9 and the trace element supplementing liquid storage unit 10 through a respective feed inlet 18; the communication modes are respectively described in the technical schemes, and are not repeated here. When a plurality of flash light illuminating incubators exist, each flash light illuminating incubator 19 respectively discharges microalgae obtained by culture through a respective discharge hole 22; the plurality of discharge ports 22 are preferably joined by piping to facilitate centralized discharge.
The microalgae cultivated by the method can enrich biomass such as protein, starch, carotenoid and the like, so that living microalgae liquid or microalgae extract is applied to farmlands, thereby promoting crop growth and increasing yield.
The invention provides an application of microalgae cultured by the method in biofertilizer.
The invention provides a biological fertilizer, wherein the effective components of the biological fertilizer preferably comprise microalgae cultured by the method. The microalgae enriched high-value microalgae biomass prepared by the method can be prepared into organic fertilizer to replace chemical fertilizer, and is applied to soil improvement and crop yield improvement.
The preparation method of the biological fertilizer is not limited, and the biological fertilizer can be prepared by adopting a mode well known to a person skilled in the art; the preparation method of the biological fertilizer in the specific application example of the invention preferably comprises the following steps:
the microalgae according to the technical scheme is preferably centrifuged to obtain the biofertilizer (algae mud). In the present invention, the rotational speed of the centrifugation is preferably 5000rpm; the time of the centrifugation is preferably 10min.
Or alternatively, the first and second heat exchangers may be,
The microalgae according to the technical scheme is preferably subjected to centrifugation, ultrasonic crushing and reduced pressure distillation to obtain the biofertilizer (microalgae extract). In the present invention, the rotational speed of the centrifugation is preferably 5000rpm; the time of the centrifugation is preferably 10min. The invention preferably carries out ultrasonic crushing on the algae mud obtained by centrifugation, and the ultrasonic crushing is preferably intermittent ultrasonic; the intermittent ultrasound is preferably ultrasound for 4s, with 4s pauses. In the ultrasonic treatment, the power is preferably 300W; the time is preferably 10min; in the ultrasonic process, the adopted ultrasonic liquid is ethanol preferably; the ethanol is preferably absolute ethanol; the volume percentage of the absolute ethyl alcohol is preferably 75%; the mass-volume ratio of the algae mud to the absolute ethyl alcohol obtained after the centrifugation is 5:1. In the present invention, the extraction mode is preferably reduced pressure distillation extraction, and the purpose of the extraction is to remove excessive ethanol; the mode of the reduced pressure distillation is not particularly limited, and may be any mode known to those skilled in the art.
The invention provides a using method of the biological fertilizer, which preferably comprises the following steps:
the biological fertilizer according to the technical scheme is diluted with water for use. In the invention, the mass-volume ratio of the biological fertilizer to the water dilution is 5:1; the means of use preferably comprises spraying; the dosage of the diluted biological fertilizer obtained by dilution is preferably 50mL/m 2; the number of applications is preferably 15.
The invention provides a device for preparing sludge hydrolysate, which comprises a sludge ultrasonic crushing unit 1 and a sludge anaerobic hydrolysis unit 2 which are communicated; the sludge ultrasonic crushing unit 1 comprises a primary sludge storage device 3 and a first stirrer 4; the sludge anaerobic hydrolysis unit 2 includes a sludge anaerobic hydrolyzer 5, an H 2O2 feed inlet 6 and a second stirrer 7, and the apparatus for preparing a sludge hydrolysate is described in detail in the foregoing technical solution, and will not be described in detail herein.
The invention provides a microalgae culture device, which comprises a sludge hydrolysate liquid storage unit 9, a microelement supplementing liquid storage unit 10 and a microalgae culture unit 11; the sludge hydrolysate liquid storage unit 9 and the trace element supplementing liquid storage unit 10 are respectively communicated with the microalgae culture unit 11; the sludge hydrolysate storage unit 9 comprises a sludge hydrolysate reservoir 12; the trace element supplementing liquid storage unit 10 comprises a trace element supplementing liquid storage 15; the microalgae culturing unit 11 comprises a feed inlet 18, a flash illumination incubator 19, a microalgae inoculation inlet 21 and a discharge outlet 22; the flash light incubator 19 includes a variable frequency light source 20, and the device for culturing microalgae is described in detail in the device for preparing sludge hydrolysate according to the foregoing technical scheme, and will not be described herein.
For further explanation of the present invention, the microalgae directional culture solution, the application thereof, the device for preparing sludge hydrolysate and the microalgae cultivation device provided by the present invention are described in detail below with reference to the accompanying drawings and examples, but they should not be construed as limiting the scope of the present invention.
The properties of the sludge used in the examples are shown in Table 1; the sludge 1, the sludge 2 and the sludge 3 are all obtained from the phenol-containing residual sludge of the Shanghai Bailong harbor sewage treatment plant.
TABLE 1 sludge characteristics
Sludge treatment VSS/TSS(％) COD(mg/L) Moisture content (%)
Sludge 1 75 200 95
Sludge 2 78 250 97
Sludge 3 80 300 99
Example 1
Preparation of sludge hydrolysate and cultivation of microalgae with reference to the apparatus shown in fig. 1 and 2:
Taking sludge 2 in table 1 to prepare hydrolysate and culturing scenedesmus obliquus (Scenedesmus obliquus), wherein the treatment method comprises the following steps:
1. Preparation of sludge hydrolysate: taking phenol-containing residual sludge of a Shanghai Bailong harbor sewage treatment plant, wherein the residual sludge is the residual sludge after the secondary biochemical treatment of industrial wastewater after heavy metal pre-removal, the heavy metal of the residual sludge reaches the agricultural sludge pollutant control standard (GB 4284-2018), namely sludge 2, standing at 25 ℃ for 6 hours, and pouring out supernatant to obtain concentrated sludge with the water content of 97%. The concentrated sludge is capped for 3d anaerobic hydrolysis at a rotation speed of 150rpm and a temperature of 37 ℃. The pH value is adjusted to 8 before anaerobic hydrolysis, the concentrated sludge is subjected to ultrasonic crushing by an ultrasonic crusher with the power of 60W and the ultrasonic frequency of 20kHz, the ultrasonic treatment is carried out for 4s, the suspension is carried out for 4s, the temperature of the primary sludge obtained after the pH value adjustment is controlled by an ice water bath is less than 50 ℃, and the treatment is carried out for 5min. H 2O2 was added after ultrasonication to 0.15mL/g VSS. Then flushing with nitrogen for 5min, sealing and reaching a strict anaerobic state. Centrifuging the sludge subjected to anaerobic hydrolysis for 3d for 10min at 8000rpm, wherein the supernatant is the sludge hydrolysate, and the sludge hydrolysate contains the following main components: 363.8Mg/L COD, 144.1Mg/L soluble protein, 73.2Mg/L soluble polysaccharide, 76.5Mg/L NH 3 -N, mg/L TN, 9.8Mg/L PO 4 -P, 13.4Mg/L TP, 40Mg/L Ca, 72Mg/L Mg, 30Mg/L Fe, 5Mg/L Mn.
2. And (3) mixing the sludge hydrolysate obtained in the step (1) with a microelement supplementing liquid containing cytokinin in a ratio of 1.2:1 to prepare the microalgae directional culture liquid. Wherein the mass percentage of cytokinin in the microelement supplementing liquid containing cytokinin is 3% of the mass of the microelement supplementing liquid, and the microelement supplementing liquid is as follows: 130g/L NaNO 3, 4.0g/L K 2HPO4, 6.5g/L MgSO 4·7H2 O, 3.5g/L CaCl 2.2H2O, 0.5g/L citric acid, 0.6g/L ferric ammonium citrate, 0.15g/L EDTANa 2, 2.0g/L Na 2CO3.
3. Culturing scenedesmus obliquus: inoculating scenedesmus obliquus (2.5X10 6 cell/mL, dry weight of 0.08 g/L) into the microalgae directional culture solution obtained in the step 2, and then culturing in an illumination incubator (25 ℃,4000Lux, flash frequency of 50Hz, light-dark ratio of 1.4:1 (namely illumination time: dark time of 14h:10 h), 20d,120 rpm), thus completing the cultivation of scenedesmus obliquus. The sludge toxicity is characterized by the inhibition rate of the luminous bacteria, and the lower the toxicity is, the lower the inhibition rate of the luminous bacteria is.
Example 2
The procedure is the same as in example 1, except that the only difference is that sludge 3 is used in this example, wherein the main components contained in the prepared sludge hydrolysate are: 364.2Mg/L COD, 145.3Mg/L soluble protein, 73.2Mg/L soluble polysaccharide, 76.5Mg/L NH 3 -N, 271.5Mg/L TN, 10Mg/L PO 4 -P, 13.5Mg/L TP, 41Mg/L Ca, 73Mg/L Mg, 31Mg/L Fe, 5.2Mg/L Mn.
Example 3
Taking sludge 3 to prepare hydrolysate and culturing scenedesmus obliquus, wherein the treatment method comprises the following steps:
1. Preparation of sludge hydrolysate: the only difference is that in this example, sludge 3 is used as the sewage, and the main components contained in the sludge hydrolysate are as follows: 365.4mg/L COD, 146.1mg/L soluble protein, 73.9mg/L soluble polysaccharide, 76.8mg/L NH 3 -N, 272.4mg/L TN, 10.2mg/L PO 4 -P, 13.8mg/L TP, 42mg/L Ca, 74mg/L Fe, 32mg/L Fe, 5.2mg/L Mn.
2. And (3) mixing the sludge hydrolysate obtained in the step (1) with a trace element supplementing liquid in a ratio of 1.5:1 to prepare the microalgae directional culture liquid. Wherein the mass percentage of cytokinin in the microelement supplementing liquid containing cytokinin is 3% of the mass of the microelement supplementing liquid, and the microelement supplementing liquid is as follows: 135g/L NaNO 3, 4.0g/L K 2HPO4, 6.5g/L MgSO 4.7H2O, 3.5g/L CaCl 2·2H2 O, 0.6g/L citric acid, 0.7g/L ferric ammonium citrate, 0.16g/L EDTANa 2, 2.0g/L Na 2CO3.
3. Culturing scenedesmus obliquus: inoculating scenedesmus obliquus (2.5X10 6 cell/mL) into the culture solution obtained in the second step, and culturing in an illumination incubator (25 ℃ C., 4000Lux, flash frequency 50Hz, light-dark ratio 1.4:1 (namely illumination time: dark time: 14h:10 h), 20d,120 rpm) to complete the cultivation of scenedesmus obliquus. The sludge toxicity is characterized by the inhibition rate of the luminous bacteria, and the lower the toxicity is, the lower the inhibition rate of the luminous bacteria is.
Example 4
Taking sludge 1 to prepare hydrolysate and culturing chlorella pyrenoidosa (Chlorellapyrenoidosa), wherein the treatment method comprises the following steps:
1. Preparation of sludge hydrolysate: the only difference is that in this example, sludge 1 is used as the sewage, and the main components contained in the sludge hydrolysate are as follows: 362.5Mg/L COD, 143.5Mg/L soluble protein, 72.5Mg/L soluble polysaccharide, 74.3Mg/L NH 3 -N, 268.6Mg/L TN, 9.8Mg/L PO 4 -P, 13.5Mg/L TP, 39Mg/L Ca, 69Mg/L Mg, 29Mg/L Fe, 4.4Mg/L Mn.
2. And (3) mixing the sludge hydrolysate obtained in the step (1) with a microelement supplementing liquid containing cytokinin in a volume ratio of 1:1 to prepare the microalgae directional culture liquid. Wherein the mass percentage of cytokinin in the microelement supplementing liquid containing cytokinin is 3% of the mass of the microelement supplementing liquid, and the microelement supplementing liquid is as follows: 136g/L NaNO 3, 4.0g/L K 2HPO4, 6.7g/L MgSO 4·7H2 O, 3.7g/L CaCl 2·2H2 O, 0.5g/L citric acid, 0.6g/L ferric ammonium citrate, 0.15g/L EDTANa 2, 2.1g/L Na 2CO3.
3. Culturing the chlorella pyrenoidosa: inoculating the chlorella pyrenoidosa (2.5X10 6 cell/mL) to the culture solution obtained in the step two, and then placing the culture solution in an illumination incubator for culture (25 ℃,4000Lux, flash frequency 50Hz, light-dark ratio of 1.4:1 (namely illumination time: dark time of 14h:10 h), 20d,120 rpm) to complete the culture of the chlorella pyrenoidosa. The sludge toxicity is characterized by the inhibition rate of the luminous bacteria, and the lower the toxicity is, the lower the inhibition rate of the luminous bacteria is.
Example 5
The only difference is that the volume ratio of the sludge hydrolysate to the trace element supplement liquid in this example is 1.2:1, as in the procedure of example 4.
Example 6
The only difference is that the volume ratio of the sludge hydrolysate to the trace element supplement liquid in this example is 1.5:1, as in the procedure of example 4.
The results of the culture in examples 4 to 6 are shown in Table 2.
TABLE 2 liquid phase and algae phase indexes in the Chlorella pyrenoidosa system cultivated by the microalgae directional culture solution prepared in examples 4-6
As is clear from Table 2, COD, sludge toxicity, TN, TP, etc. were remarkably removed after culturing Chlorella pyrenoidosa in the sludge hydrolysate. Along with the increase of the proportion of the sludge hydrolysate, the cell density of the chlorella pyrenoidosa is gradually increased, and the proteins, the polysaccharides, the carotenoids and the like are obviously increased, which shows that the chlorella pyrenoidosa can absorb and utilize the nutrient substances in the sludge hydrolysate to grow and simultaneously obviously enrich the value-added biomass and remove the sludge toxicity.
Example 7
Taking excess sludge 2 to prepare hydrolysate and culturing Chlorella terrestris (Chlorococcum humicola), wherein the treatment method comprises the following steps:
1. Preparation of sludge hydrolysate: the same procedure as in example 1 was followed, except that the present example was allowed to stand for 8 hours, wherein the main components contained in the sludge hydrolysate were: 364.2mg/L COD, 144.0mg/L soluble protein, 73.2mg/L soluble polysaccharide, 76.3mg/L NH 3 -N, 271.4mg/L TN, 10.0mg/L PO 4 -P, 14.1mg/L TP, 40mg/L Ca, 72mg/L Fe, 31mg/L Fe, 5.2mg/L Mn.
2. And (3) mixing the sludge hydrolysate obtained in the step (1) with a microelement supplementing liquid containing cytokinin in a ratio of 1.5:1 to prepare the microalgae directional culture liquid. Wherein the mass percentage of cytokinin in the microelement supplementing liquid containing cytokinin is 3% of the mass of the microelement supplementing liquid, and the microelement supplementing liquid is as follows: 130g/L NaNO 3, 4.0g/L K 2HPO4, 6.5g/L MgSO 4·7H2 O, 3.5g/L CaCl 2·2H2 O, 0.5g/L citric acid, 0.6g/L ferric ammonium citrate, 0.15g/L EDTANa 2, 2.0g/L Na 2CO3.
3. Culturing Chlorella oxytoca: inoculating Chlorella terrestris (2.5X10 6 cell/mL) to the culture solution obtained in the step 2, and then placing the culture solution in an illumination incubator for cultivation (25 ℃,4000Lux, flash frequency 50Hz, light-dark ratio of 1.4:1 (namely illumination time: dark time: 14h:10 h), 20d,120 rpm) to complete the cultivation of Chlorella terrestris. The sludge toxicity is characterized by the inhibition rate of the luminous bacteria, and the lower the toxicity is, the lower the inhibition rate of the luminous bacteria is. The culture results are shown in Table 5.
Example 8
The procedure is the same as in example 7, except that in this example, H 2O2 0.1.1 mL/g VSS is added after the ultrasonic disruption of the sludge.
Example 9
The procedure is the same as in example 7, except that in this example, H 2O2 0.2.2 mL/g VSS is added after the ultrasonic disruption of the sludge.
The results of the culture in examples 7 to 9 are shown in Table 3.
TABLE 3 liquid phase and algae phase indexes in Chlorella terrestris culture systems by microalgae directional culture solution prepared in examples 7-9
As is clear from the experimental data shown in Table 3, when the amount of H 2O2 added was 0.15mL/gVSS, the obtained sludge hydrolysate was mixed with the trace element supplement solution to prepare a microalgae culture solution for cultivating Chlorella terrestris, which can obtain the highest microalgae cell density of 2 8×106 cells/mL, and simultaneously enrich the biomass such as protein, polysaccharide, carotenoid, etc. The addition of 0.15mL/g VSS H 2O2 is most beneficial to promoting cell cracking to release nutrient substances, so that the nutrient substances can be fully anaerobically hydrolyzed in an anaerobic stage, and the sludge hydrolysate with rich nutrient substances is obtained. Therefore, the growth of the microalgae can be obviously promoted when the microalgae are cultivated, and the sludge toxicity is reduced through the symbiosis of the microalgae and bacteria.
Example 10
Taking excess sludge 3 to prepare hydrolysate and culturing scenedesmus obliquus, wherein the treatment method comprises the following steps:
1. Preparation of sludge hydrolysate: the only difference is that in this example, sludge 3 is used as the sewage, and the main components contained in the sludge hydrolysate are as follows: 366.5Mg/L COD, 146.8Mg/L soluble protein, 74.8Mg/L soluble polysaccharide, 82.5Mg/L NH 3 -N, 276.5Mg/L TN, 10.2Mg/L PO 4 -P, 14.2Mg/L TP, 43Mg/L Ca, 75Mg/L Mg, 33Mg/L Fe, 5.0Mg/L Mn.
2. And (3) mixing the sludge hydrolysate obtained in the step (1) with a microelement supplementing liquid containing cytokinin in a ratio of 1.5:1 to prepare the microalgae directional culture liquid. Wherein the mass percentage of cytokinin in the microelement supplementing liquid containing cytokinin is 3% of the mass of the microelement supplementing liquid, and the microelement supplementing liquid is as follows: 135g/L NaNO 3, 4.0g/L K 2HPO4, 6.5g/L MgSO 4·7H2 O, 3.5g/L CaCl 2·2H2 O, 0.6g/L citric acid, 0.6g/L ferric ammonium citrate, 0.15g/L EDTANa 2, 2.0g/L Na 2CO3.
3. Culturing scenedesmus obliquus: inoculating scenedesmus obliquus (2.5X10 6 cell/mL) into the culture solution obtained in the step 2, and culturing in an illumination incubator (25 ℃ C., 4000Lux, flash frequency 50Hz, light-dark ratio 1.4:1 (namely illumination time: dark time: 14h:10 h), 20d,120 rpm) to complete the cultivation of scenedesmus obliquus. The sludge toxicity is characterized by the inhibition rate of the luminous bacteria, and the lower the toxicity is, the lower the inhibition rate of the luminous bacteria is.
Example 11
The procedure is as in example 10, with the only difference that 2% cytokinin is added to the trace element addition solution in this example.
Example 12
The procedure is as in example 10, with the only difference that in this example 4% cytokinin is added to the trace element addition solution.
The results of the culture in examples 10 to 12 are shown in Table 4.
TABLE 4 liquid phase and algal phase indexes in a Scenedesmus obliquus system cultivated by using the microalgae directional culture solution prepared in examples 10-12
From the experimental data shown in table 4, it is clear that the addition of 3% cytokinin to the trace element supplementing liquid for cultivating scenedesmus obliquus can stimulate the growth of scenedesmus obliquus, and absorb and utilize the nutrients in the sludge extracting liquid, so that scenedesmus obliquus can produce high-content biomass such as protein, polysaccharide, carotenoid and the like.
Comparative example 1
The only difference is that in this example, sludge 1 is used, wherein the main components contained in the sludge hydrolysate are as follows: 361.5Mg/L COD, 143.0Mg/L soluble protein, 72.5Mg/L soluble polysaccharide, 71.6Mg/L NH 3 -N, 267.5Mg/L TN, 9.8Mg/L PO 4 -P, 12.8Mg/L TP, 39Mg/L Ca, 69Mg/L Mg, 29Mg/L Fe, 4.4Mg/L Mn.
The results of the microalgae directional culture liquids of example 1, example 2 and comparative example 1 are shown in Table 5. TABLE 5 liquid and algal phase indicators in a Scenedesmus obliquus culture System with the microalgae directional culture solutions prepared in example 1, example 2 and comparative example 1
As is clear from the experimental data shown in Table 5, the sludge with VSS/TSS > 70% was used to significantly reduce the sludge content of the sludge, and after microalgae cultivation, the protein, polysaccharide and carotenoid content in the microalgae could be significantly increased, and the sludge toxicity was eliminated by the algae symbiotic system.
Comparative example 2
The same as in example 3, except that the concentrated sludge was not pretreated, and was directly subjected to 3d anaerobic hydrolysis by capping after being purged with nitrogen for 5 minutes at a rotation speed of 150rpm and a temperature of 37℃to achieve a strictly anaerobic state.
Comparative example 3
The procedure was as in example 3, except that the pH value was adjusted to 11 before anaerobic hydrolysis of the concentrated sludge, and after the anaerobic condition was achieved by purging with nitrogen for 5 minutes, the anaerobic hydrolysis was conducted by capping at a rotation speed of 150rpm and a temperature of 37℃for 3 d.
The results of the cultivation of example 4, comparative example 2 and comparative example 3 are shown in Table 6.
TABLE 6 liquid and algal phase index in a Scenedesmus obliquus culture System
As shown in experimental data recorded in Table 6, the preparation method can fully release organic matters in sludge, can obtain sludge hydrolysate rich in nutrients such as carbon, nitrogen and phosphorus after anaerobic hydrolysis, can remarkably improve the contents of protein, polysaccharide and carotenoid in microalgae after microalgae cultivation, and can remove the toxicity of the sludge by using a algae symbiotic system.
Application example 1
Collecting chlorella pyrenoidosa algae liquid after 20d culture in example 6, taking out a proper amount of algae liquid, centrifuging (5000 rpm,10 min), removing supernatant, collecting algae mud, adding absolute ethyl alcohol into the algae mud, shaking, intermittently performing ultrasonic crushing for 10min (300W, ultrasonic 4s, interval 4 s) in an ice water bath to obtain crushed liquid, wherein the mass volume ratio of the algae mud to the absolute ethyl alcohol is 5:1, the volume percent of the absolute ethyl alcohol is 75%, shaking the crushed liquid for 1h, distilling under reduced pressure to remove the ethyl alcohol, adding tap water, shaking to obtain a microalgae extract, and wherein the mass volume ratio of the microalgae extract to the tap water is 5:1. The microalgae extract is applied to a pot plant planted with 3 cucumber seeds, the microalgae extract is fertilized once for 3d Shi Zao before the cucumber seeds are sown, the microalgae extract is applied once for 5d before the cucumber flowers are planted, the microalgae extract is fertilized once for 3d after the cucumber flowers are planted, and the microalgae extract is fertilized 15 times for 50mL/m 2 each time. Observing and recording cucumber growth vigor every day, and when the cucumber seedlings are all grown to 4-5 cm in soil, keeping 1 plant with the best growth vigor (namely, the plant has straight stems and green leaves and has no insect damage); when the plant grows to 30d, recording plant height, leaf number, stem diameter and the like; when the plant grows to 45d (flowering period), the number of buds, plant height, leaf number, stem diameter and the like are recorded.
Application example 2
The procedure was as in application example 1, except that the obtained algae puree was mixed with tap water to obtain a living microalgae algae liquid, wherein the mass-to-volume ratio of the algae puree to tap water was 5:1, and the mixture was applied to a pot plant in which cucumber was planted.
Application example 3
The same procedure as in application example 1 was repeated except that tap water was applied only to the pot plants in which cucumbers were planted, and no algae liquid was added.
The results of the culture in application examples 1 to 3 are shown in Table 7.
TABLE 7 application examples 1-3 preparation of cucumber growth index in algae liquid culture cucumber System
From the experimental data shown in Table 7, it is clear that the growth of cucumber is better after the application of living microalgae algae liquid or microalgae extract than after the application of tap water, especially when microalgae extract is applied, the plant height can reach 62cm and the number of flower buds reaches 18, which means that the biomass such as protein, starch, carotenoid and the like enriched in microalgae can be used as biofertilizer to promote the growth of crops and increase the yield.
As is clear from the above examples, the invention eliminates the toxicity of the sludge by forming a zoobacterial symbiotic system by microalgae and bacteria in the sludge hydrolysate, and reduces the biological toxicity of the sludge from 80% to only about 10%; the components in the microelement supplementing liquid containing cytokinin can meet the growth requirement of microalgae in anaerobic hydrolysate, and provide microelement for the growth of microalgae; cytokinin is a plant hormone, can promote green algae to synthesize biomass such as glutamic acid, starch, pigment and the like, and can supplement microelement supplementing liquid containing cytokinin to promote the growth of green algae and obtain value-added biomass such as protein, polysaccharide, carotenoid and the like; therefore, the culture solution can remarkably improve the yield of microalgae value-added biomass by utilizing the sludge hydrolysate, cytokinin and trace element supplementing liquid, and a large amount of surplus sludge can be utilized by utilizing the sludge hydrolysate, so that the cost is relatively low, and the surplus sludge is recycled. In addition, the invention provides a preparation method of the sludge hydrolysate, which can better break the sludge and release the nutrient substances contained in the sludge. The invention also provides a microalgae culture method, which can save microalgae culture cost, capture biomass value-added products of glutamic acid proteins up to 400mg/L, starch 200mg/L and carotenoid 5mg/L, realize ecological recycling and reduction utilization of residual sludge, and capture microalgae biomass as a biofertilizer which can be applied to agricultural production in farmlands, promote crop growth and increase yield.
While the invention has been described in terms of preferred embodiments, it is not intended to be limited thereto, but rather to enable any person skilled in the art to make various changes and modifications without departing from the spirit and scope of the present invention, which is therefore to be limited only by the appended claims.

Claims (5)

1. The microalgae directional culture solution is characterized by comprising a sludge hydrolysate and a microelement supplementing solution containing cytokinin; the trace element supplementing liquid comprises the following components in mass: 120-150 g/L NaNO 3, 3-5 g/L K 2HPO4, 5-8 g/L MgSO 4·7H2 O, 2-5 g/L CaCl 2·2H2 O, 0.4-0.7 g/L citric acid, 0.5-0.8 g/L ferric ammonium citrate, 0.1-0.2 g/L EDTANa 2, 1-3 g/L Na 2CO3;
The mass percentage of cytokinin in the microelement supplementing liquid containing cytokinin is 2% -3% of the mass of the microelement supplementing liquid;
The volume ratio of the sludge hydrolysate to the microelement supplementing liquid containing cytokinin is (1.2-1.5): 1;
the microalgae are one or more of Chlorella pyrenoidosa, scenedesmus obliquus and Chlorella terrestris;
the preparation of the sludge hydrolysate comprises the following steps:
Standing the sludge to obtain concentrated sludge;
Sequentially adjusting the pH value, carrying out ultrasonic crushing and anaerobic hydrolysis on the concentrated sludge to obtain sludge hydrolysate; the pH value of the primary sludge obtained after the adjustment is more than or equal to 7;
the sludge is the excess sludge of an industrial wastewater treatment plant; the VSS/TSS of the sludge is more than 70%; the standing time is 4-8 hours;
The ultrasonic crushing further comprises adding H 2O2 into the intermediate sludge obtained after ultrasonic crushing; the addition amount of the H 2O2 is 0.1-0.15 mL/gVSS.
2. The microalgae directional culture liquid of claim 1, wherein the cytokinin comprises one or more of kinetin, zeatin and 6-aminobenzyl purine.
3. The microalgae directional culture liquid of claim 1, wherein the ultrasound is intermittent ultrasound; the intermittent ultrasound is ultrasound for 4s, and pausing for 4s; in the ultrasonic treatment, the ultrasonic frequency is 10-30 kHz, the power is 50-80W, the time is 5min, and the temperature of the primary sludge obtained after the pH value is regulated is less than 50 ℃.
4. The use of the microalgae directional culture solution according to any one of claims 1-3 in directional culture of biomass-enriched microalgae, characterized in that the microalgae are one or more of chlorella pyrenoidosa, scenedesmus obliquus and chlorella terrestris.
5. A method for directionally culturing and enriching biomass microalgae, which is characterized by comprising the following steps:
The culture medium according to any one of claims 1 to 3, which is inoculated with microalgae and cultured.
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