CN113493246A - Method for reducing total nitrogen and total phosphorus in aquaculture wastewater by utilizing microalgae photobiological reaction - Google Patents

Method for reducing total nitrogen and total phosphorus in aquaculture wastewater by utilizing microalgae photobiological reaction Download PDF

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CN113493246A
CN113493246A CN202110680630.8A CN202110680630A CN113493246A CN 113493246 A CN113493246 A CN 113493246A CN 202110680630 A CN202110680630 A CN 202110680630A CN 113493246 A CN113493246 A CN 113493246A
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microalgae
wastewater
culture
total nitrogen
total phosphorus
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赵璐
王松
张新明
胡晓文
梁青
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Rizhao Polytechnic
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    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00—Biological treatment of water, waste water, or sewage
    • C02F3/32—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae
    • C02F3/322—Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae use of algae
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/12—Unicellular algae; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
    • C12N11/08—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer
    • C12N11/082—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a synthetic polymer obtained by reactions only involving carbon-to-carbon unsaturated bonds
    • C12N11/084—Polymers containing vinyl alcohol units
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
    • C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00—Nature of the contaminant
    • C02F2101/10—Inorganic compounds
    • C02F2101/105—Phosphorus compounds
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00—Nature of the contaminant
    • C02F2101/10—Inorganic compounds
    • C02F2101/16—Nitrogen compounds, e.g. ammonia
    • C—CHEMISTRY; METALLURGY
    • C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
    • C02F2103/20—Nature of the water, waste water, sewage or sludge to be treated from animal husbandry

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Abstract

The invention belongs to the technical field of wastewater treatment, and particularly relates to a method for reducing total nitrogen and total phosphorus in aquaculture wastewater by utilizing microalgae photobiological reaction, which comprises the following steps: selecting microalgae species, sequentially carrying out activation and amplification culture, inoculating the microalgae species to a photobioreactor for high-density culture until the logarithmic growth phase of cells, and then carrying out centrifugal collection and washing on the microalgae to prepare a microalgae suspension; adding activated sludge into the microalgae suspension to prepare a mixed solution; adding the mixed solution into a carrier solution, oscillating at constant temperature to uniformly disperse the mixed solution, and dropwise adding a calcium chloride solution to precipitate solidified microspheres until the solidified microspheres are not precipitated; after pretreatment of the breeding wastewater, adding curing microspheres for treatment; and collecting the solidified microspheres in the culture wastewater removal liquid. The method can ensure the stability of the plasmid, reduce the mutation probability, and can be repeatedly used for multiple times, thereby improving the removal rate of nitrogen and phosphorus and ensuring the purification effect.

Description

Method for reducing total nitrogen and total phosphorus in aquaculture wastewater by utilizing microalgae photobiological reaction
Technical Field
The invention belongs to the technical field of wastewater treatment, and particularly relates to a method for reducing total nitrogen and total phosphorus in aquaculture wastewater by utilizing microalgae photobiological reaction.
Background
In recent years, the scale of aquaculture industry is gradually reduced from small to scattered, and the aquaculture industry is developed towards large-scale and intensification, and the large-scale aquaculture has the advantages of cost saving, optimized management, yield increase and the like, but due to the defecation characteristic of livestock and poultry in the mode, a large amount of excrement wastewater containing pollutants such as nitrogen, phosphorus, antibiotics and the like is generated in the aquaculture process, and in order to save cost, a direct discharge or rough treatment method is usually adopted to treat the wastewater, so that the environment is seriously affected, and therefore, the research on an efficient, low-cost, green and environment-friendly wastewater treatment method is urgently needed by enterprises and ecological environments for treating the livestock and poultry wastewater, purifying and protecting water resources.
Microalgae are unicellular microorganisms widely existing in various water environments, and a technology for purifying wastewater by using microalgae has attracted attention. The culture wastewater contains a large amount of nitrogen, phosphorus and organic matters, so that the requirements of microalgae on a nitrogen source and a carbon source can be met, the content of nitrogen and phosphorus in water is effectively reduced, and meanwhile, the microalgae can also adsorb or degrade pollutants such as heavy metals, antibiotics and the like in the wastewater, so that the purification of the wastewater is realized. In the prior art, suspended algae is often adopted to remove nitrogen and phosphorus in wastewater, but the method cannot ensure the plasmid stability of the algae, has high mutation probability, cannot realize recycling, and is not ideal enough in nitrogen and phosphorus removal rate.
Disclosure of Invention
In order to solve the problem that the removal rate of nitrogen and phosphorus by algae is not ideal in the prior art, the invention provides a method for reducing total nitrogen and total phosphorus in aquaculture wastewater by utilizing microalgae photobiological reaction, and the removal rate of nitrogen and phosphorus in the aquaculture wastewater is improved.
The invention provides a method for reducing total nitrogen and total phosphorus in culture wastewater by utilizing microalgae photobiological reaction, which comprises the following steps:
(1) selecting microalgae species, sequentially carrying out activation and amplification culture, inoculating the microalgae species to a photobioreactor for high-density culture until the logarithmic growth phase of cells, and then carrying out centrifugal collection and washing on the microalgae to prepare a microalgae suspension;
(2) adding activated sludge into the microalgae suspension to prepare a mixed solution;
(3) adding the mixed solution into a carrier solution, oscillating at constant temperature to uniformly disperse the mixed solution, and dropwise adding a calcium chloride solution to precipitate solidified microspheres until the solidified microspheres are not precipitated;
(4) after pretreatment of the breeding wastewater, adding curing microspheres for treatment;
(5) and collecting the solidified microspheres in the culture wastewater removal liquid.
Further, in the step (1), the microalgae is marine nannochloropsis oceanica, the preservation unit is China general microbiological culture Collection center, the address is No. 3 Siro No. 1 of the Chaozhou, the republic of Beijing, the south-facing-Yang district, the preservation time is 10-23 days of 2020, the preservation number is CGMCC No.20713, and the reference biological materials (strains) are as follows: LAMB204, the proposed classification nomenclature: marine Nannochloropsis oceanica.
Further, in the step (1), the high-density culture conditions are as follows: the illumination intensity is 4000lux-7000lux, the light-dark time ratio is 14 h: 10h-16 h: 8h, CO2The gas ventilation amount is 0.01-0.2VVm, pH is 5.5-6.5, culture temperature is 18-30 deg.C, and culture time is 2-4 days.
Further, in the step (1), the bacterial density of the microalgae after high-density culture reaches 107-109CFU/mL。
Further, in the step (1), the centrifugation speed is 3000-4000rpm, and the centrifugation time is 10-15 min.
Further, in the step (2), the adding amount of the activated sludge is 20-30g/L in dry weight.
Further, in the step (3), the carrier solution is a sodium alginate-polyvinyl alcohol solution. The carrier can absorb nutrients in the culture wastewater, is beneficial to the growth of algae, ensures the repeated utilization of the solidified microspheres, can fade the color of the wastewater under the condition of not influencing the growth of other cells, and improves the clarity of the wastewater.
Further, in the step (3), the concentration of calcium chloride is 3.0-4.5 wt%.
Further, in the step (4), the pretreatment step is: aerating the culture wastewater in an aeration tank for 20-40min, and precipitating for 20-40 min.
Further, in the step (4), the addition amount of the cured microspheres is 3-5g/L in terms of dry weight.
The invention has the beneficial effects that:
according to the method for reducing total nitrogen and total phosphorus in the aquaculture wastewater by utilizing the microalgae photobiological reaction, the microalgae are mixed with the activated sludge, so that the enrichment and absorption capacity of nitrogen and phosphorus elements in the wastewater is improved, and the purification effect is ensured; the immobilized microspheres are formed by being placed in a carrier solution, so that the stability of plasmids can be better guaranteed, the mutation probability is reduced, the immobilized microspheres can be repeatedly utilized for multiple times, and the removal rate of nitrogen and phosphorus is greatly improved. In addition, the method is simple and efficient, can quickly prepare a large amount of high-quality cured microspheres, and has low cost and obvious effect.
Detailed Description
In order to make those skilled in the art better understand the technical solutions in the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. 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.
Example 1
Taking 15L of wastewater from a waterfowl breeding wastewater factory, and removing suspended matters and large-scale plankton by conventional isolation, wherein the total nitrogen content is 400mg/L, the total phosphorus content is 50mg/L, and the pH value is 3.5.
A method for reducing total nitrogen and total phosphorus in culture wastewater by utilizing microalgae photobiological reaction comprises the following steps:
(1) selecting marine nannochloropsis oceanica, sequentially carrying out activation and amplification culture, inoculating the marine nannochloropsis oceanica to a photobioreactor for high-density culture until the logarithmic growth phase of cells, then carrying out centrifugal collection on the microalgae, wherein the centrifugal rotation speed is 3000rpm, the centrifugal time is 15min, and then washing the microalgae to prepare a microalgae suspension;
the marine nannochloropsis oculata preservation unit is the common microorganism center of China Committee for culture Collection of microorganisms, the address is No. 3 of No. 1 Xilu-Chen-Yang district of Beijing, the preservation time is 10 months 23 days in 2020, the preservation number is CGMCC No.20713, and the reference biological materials (strains) are as follows: LAMB204, the proposed classification nomenclature: marine Nannochloropsis oceanica;
the high-density culture conditions are as follows: the illumination intensity is 4000 luxluxlux, the light-dark time ratio is 16 h: 8h, CO2The gas ventilation rate is 0.1VVm, the pH is 5.5, the culture temperature is 20 ℃, and the culture time is 3 days; the bacterial density of the microalgae after high-density culture reaches 107-109CFU/mL。
(2) 20g/L (dry weight) of activated sludge was added to the microalgae suspension to prepare a mixed solution.
(3) Adding the mixed solution into a sodium alginate-polyvinyl alcohol solution, oscillating at constant temperature to uniformly disperse the mixed solution, and dropwise adding a 3.0 wt% calcium chloride solution to separate out the solidified microspheres until the microspheres are not separated out.
(4) After the breeding wastewater is pretreated, 3g/L (dry weight) of curing microspheres are added for treatment; the pretreatment steps are as follows: adjusting the pH value of the aquaculture wastewater to 5.5, then aerating in an aeration tank for 25min, and precipitating for 25 min.
(5) And collecting the solidified microspheres in the culture wastewater removal liquid.
And (3) carrying out nitrogen and phosphorus measurement on the treated aquaculture wastewater, wherein the total nitrogen content is 27mg/L, the removal rate is 93.25%, the total phosphorus content is 5.4mg/L, and the removal rate is 89.2%, and the livestock and poultry aquaculture pollutant discharge standard GB 18596-2001 is met.
Example 2
20L of wastewater is taken from a waterfowl breeding wastewater factory, and after suspended matters and large plankton are removed by conventional separation, the total nitrogen content is 400mg/L, the total phosphorus content is 50mg/L, and the pH value is 3.5.
A method for reducing total nitrogen and total phosphorus in culture wastewater by utilizing microalgae photobiological reaction comprises the following steps:
(1) selecting marine nannochloropsis oceanica, sequentially carrying out activation and amplification culture, inoculating the marine nannochloropsis oceanica to a photobioreactor for high-density culture until the logarithmic growth phase of cells, then carrying out centrifugal collection on the microalgae, wherein the centrifugal rotation speed is 4000rpm, the centrifugal time is 12min, and then washing the microalgae to prepare a microalgae suspension;
the marine nannochloropsis oculata preservation unit is the common microorganism center of China Committee for culture Collection of microorganisms, the address is No. 3 of No. 1 Xilu-Chen-Yang district of Beijing, the preservation time is 10 months 23 days in 2020, the preservation number is CGMCC No.20713, and the reference biological materials (strains) are as follows: LAMB204, the proposed classification nomenclature: marine Nannochloropsis oceanica;
the high-density culture conditions are as follows: the illumination intensity is 6000lux, the light-dark time ratio is 15 h: 9h, CO2The gas ventilation amount is 0.2VVm, the pH is 6.5, the culture temperature is 18-30 ℃, and the culture time is 4 days; the bacterial density of the microalgae after high-density culture reaches 107-109CFU/mL。
(2) 30g/L (dry weight) of activated sludge was added to the microalgae suspension to prepare a mixed solution.
(3) Adding the mixed solution into a sodium alginate-polyvinyl alcohol solution, oscillating at constant temperature to uniformly disperse the mixed solution, and dropwise adding a 4.5 wt% calcium chloride solution to separate out the solidified microspheres until the microspheres are not separated out.
(4) Pretreating the breeding wastewater, and adding 5g/L (dry weight) of curing microspheres for treatment; the pretreatment steps are as follows: adjusting the pH value of the aquaculture wastewater to 6.5, then introducing the aquaculture wastewater into an aeration tank for aeration for 40min, and precipitating for 40 min.
(5) And collecting the solidified microspheres in the culture wastewater removal liquid.
And (3) carrying out nitrogen and phosphorus measurement on the treated aquaculture wastewater, wherein the total nitrogen content is 31mg/L, the removal rate is 92.25%, the total phosphorus content is 5.2mg/L, and the removal rate is 89.6%, and the livestock and poultry aquaculture pollutant discharge standard GB 18596-2001 is met.
Comparative example 1
According to the method for reducing total nitrogen and total phosphorus in the culture wastewater by utilizing the microalgae photo-biological reaction, activated sludge is not added into the microalgae suspension, and other preparation methods are the same as those in the example 2.
The nitrogen and phosphorus determination is carried out on the treated aquaculture wastewater, the total nitrogen content is 54mg/L, the removal rate is 86.5%, the total phosphorus content is 7.1mg/L, and the removal rate is 85.8%, compared with the example 2, the comparative example has lower total nitrogen and total phosphorus removal rate and slightly poor wastewater treatment effect.
Comparative example 2
According to the method for reducing total nitrogen and total phosphorus in the aquaculture wastewater by utilizing the microalgae photo-biological reaction, activated sludge is not added into the microalgae suspension, solidified microspheres are not prepared, the microalgae suspension is directly added into the wastewater for treatment, and other preparation methods are the same as those in example 2.
The nitrogen and phosphorus determination is carried out on the treated aquaculture wastewater, the total nitrogen content is 167mg/L, the removal rate is 58.25%, the total phosphorus content is 19.4mg/L, and the removal rate is 61.2%, compared with the example 2, the comparative example has extremely low total nitrogen and total phosphorus removal rate and extremely poor wastewater treatment effect.
Although the present invention has been described in detail by way of preferred embodiments, the present invention is not limited thereto. Various equivalent modifications or substitutions can be made on the embodiments of the present invention by those skilled in the art without departing from the spirit and scope of the present invention, and these modifications or substitutions are within the scope of the present invention/any person skilled in the art can easily conceive of the changes or substitutions within the technical scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.

Claims (10)

1. A method for reducing total nitrogen and total phosphorus in culture wastewater by utilizing microalgae photobiological reaction is characterized by comprising the following steps:
(1) selecting microalgae species, sequentially carrying out activation and amplification culture, inoculating the microalgae species to a photobioreactor for high-density culture until the logarithmic growth phase of cells, and then carrying out centrifugal collection and washing on the microalgae to prepare a microalgae suspension;
(2) adding activated sludge into the microalgae suspension to prepare a mixed solution;
(3) adding the mixed solution into a carrier solution, oscillating at constant temperature to uniformly disperse the mixed solution, and dropwise adding a calcium chloride solution to precipitate solidified microspheres until the solidified microspheres are not precipitated;
(4) after pretreatment of the breeding wastewater, adding curing microspheres for treatment;
(5) and collecting the solidified microspheres in the culture wastewater removal liquid.
2. The method for reducing total nitrogen and total phosphorus in aquaculture wastewater by utilizing microalgae photobiological reaction as claimed in claim 1, wherein the microalgae is marine nannochloropsis oculata, the preservation unit is the common microorganism center of China Committee for culture Collection of microorganisms, the address is No. 3 of No. 1 Siro-Chen-Su-Lu-1 of the area facing the sun in Beijing, the preservation time is 10-23 days in 2020 year, the preservation number is CGMCC No.20713, and the reference biological materials (strains) are as follows: LAMB204, the proposed classification nomenclature: marine Nannochloropsis oceanica.
3. The method for reducing total nitrogen and total phosphorus in aquaculture wastewater by utilizing microalgae photobiological reaction as claimed in claim 1, wherein in the step (1), the high-density culture conditions are as follows: the illumination intensity is 4000lux-7000lux, the light-dark time ratio is 14 h: 10h-16 h: 8h, CO2The gas ventilation amount is 0.01-0.2VVm, pH is 5.5-6.5, culture temperature is 18-30 deg.C, and culture time is 2-4 days.
4. The method for reducing total nitrogen and total phosphorus in aquaculture wastewater according to claim 1, wherein the method comprises the step of reducing total nitrogen and total phosphorus in aquaculture wastewater by utilizing microalgae photobiological reactionCharacterized in that in the step (1), the bacterial density of the microalgae after high-density culture reaches 107-109CFU/mL。
5. The method for reducing total nitrogen and total phosphorus in aquaculture wastewater by utilizing microalgae photobioreactor as claimed in claim 1, wherein in the step (1), the centrifugation rotation speed is 3000-4000rpm, and the centrifugation time is 10-15 min.
6. The method for reducing total nitrogen and total phosphorus in aquaculture wastewater by utilizing microalgae photobioreactor as claimed in claim 1, wherein in the step (2), the addition amount of activated sludge is 20-30g/L in dry weight.
7. The method for reducing total nitrogen and total phosphorus in aquaculture wastewater by utilizing microalgae photobioreactor as claimed in claim 1, wherein in the step (3), the carrier solution is sodium alginate-polyvinyl alcohol solution.
8. The method for reducing total nitrogen and total phosphorus in aquaculture wastewater by utilizing microalgae photobioreactor as claimed in claim 1, wherein in the step (3), the concentration of calcium chloride is 3.0-4.5 wt%.
9. The method for reducing total nitrogen and total phosphorus in aquaculture wastewater by utilizing microalgae photobiological reaction as claimed in claim 1, wherein in the step (4), the pretreatment step comprises the following steps: aerating the culture wastewater in an aeration tank for 20-40min, and precipitating for 20-40 min.
10. The method for reducing total nitrogen and total phosphorus in aquaculture wastewater by utilizing microalgae photobioreactor as claimed in claim 1, wherein in the step (4), the addition amount of the solidified microspheres is 3-5g/L in terms of dry weight.
CN202110680630.8A 2021-06-18 2021-06-18 Method for reducing total nitrogen and total phosphorus in aquaculture wastewater by utilizing microalgae photobiological reaction Withdrawn CN113493246A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116152445A (en) * 2023-04-17 2023-05-23 日照职业技术学院 Microalgae distribution judging method of microalgae culture pond

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116152445A (en) * 2023-04-17 2023-05-23 日照职业技术学院 Microalgae distribution judging method of microalgae culture pond

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Application publication date: 20211012