CN110723824A - Preparation of low-temperature denitrification microbial inoculum and method for enhancing denitrification efficiency of constructed wetland - Google Patents

Preparation of low-temperature denitrification microbial inoculum and method for enhancing denitrification efficiency of constructed wetland Download PDF

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CN110723824A
CN110723824A CN201911011186.XA CN201911011186A CN110723824A CN 110723824 A CN110723824 A CN 110723824A CN 201911011186 A CN201911011186 A CN 201911011186A CN 110723824 A CN110723824 A CN 110723824A
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temperature
low
denitrification
microbial inoculum
denitrifying
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周晶
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Qufu Normal University
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    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00Biological treatment of water, waste water, or sewage
    • C02F3/34Biological treatment of water, waste water, or sewage characterised by the microorganisms used
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00Biological treatment of water, waste water, or sewage
    • C02F3/30Aerobic and anaerobic processes
    • C02F3/302Nitrification and denitrification treatment
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F3/00Biological treatment of water, waste water, or sewage
    • C02F3/32Biological treatment of water, waste water, or sewage characterised by the animals or plants used, e.g. algae
    • CCHEMISTRY; METALLURGY
    • C02TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02FTREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
    • C02F2101/00Nature of the contaminant
    • C02F2101/10Inorganic compounds
    • C02F2101/16Nitrogen compounds, e.g. ammonia

Abstract

The invention discloses a method for preparing a low-temperature denitrification microbial inoculum and enhancing the denitrification efficiency of an artificial wetland, which comprises the processes of enriching and domesticating a low-temperature nitrifying flora and enriching and domesticating a low-temperature aerobic denitrifying flora.

Description

Preparation of low-temperature denitrification microbial inoculum and method for enhancing denitrification efficiency of constructed wetland
Technical Field
The invention relates to the technical field of constructed wetland treatment, in particular to a preparation method of a low-temperature denitrification microbial inoculum and a method for enhancing the denitrification efficiency of the constructed wetland.
Background
The constructed wetland contains a large amount of aerobic microorganisms and facultative microorganisms, and in the wetland with composite vertical flow, the number of bacteria on the surface layer is higher than that on the lower layer, and the microbial community is obviously layered. The number and community structure of microorganisms in the artificial wetland are greatly changed along with different seasons, and the artificial wetland system has strong degradation capability on organic matters. Insoluble organic matters in the sewage are intercepted through the filtration and precipitation of the matrix, the removal effect of the matrix on the organic matters is reduced due to adsorption saturation, but microorganisms attached to the surface of the matrix in large quantity can complete the removal of the organic matters through a metabolic process.
However, when the water temperature is reduced to below 10 ℃, the microbial activity is greatly reduced, the biochemical treatment effect is reduced, measures such as load reduction, hydraulic retention time extension, heat preservation and the like are commonly adopted in engineering to enable the effluent quality in winter to reach the standard, but the measures increase the operation cost and have unstable treatment effect.
Disclosure of Invention
The invention aims to provide a preparation method of a low-temperature denitrification microbial inoculum and a method for enhancing denitrification efficiency of an artificial wetland.
In order to achieve the above purpose of the present invention, the following technical solutions are adopted:
the invention comprises the following steps:
(1) enrichment and domestication of low-temperature nitrifying bacteria: performing low-temperature acclimation on a nitrobacteria culture medium according to a temperature gradient of 30 ℃ → 25 ℃ → 20 → 15 ℃ → 13 → 10 ℃, and culturing and enriching in a mixed solution containing trace elements and nitrate and nitrogen in a 250mL shake flask under aerobic conditions, wherein the dissolved oxygen is 0.1-5 mg/L, the pH is 7.0-9.0, the shaking table rotating speed is 150rpm, the period is 12h per week, and the water discharge rate is about 90%;
(2) enrichment and domestication of low-temperature aerobic denitrification flora: activating aerobic denitrifying bacteria at the normal temperature at 30 ℃, and performing low-temperature acclimation on the activated aerobic denitrifying bacteria under the aerobic condition according to the temperature gradient of 30 ℃→ 20 → 15 ℃ → 10 ℃;
(3) inoculating the low-temperature nitrifying bacteria and the denitrifying bacteria into a fermentation culture medium for amplification culture to obtain the low-temperature aerobic denitrifying bacteria agent.
Specifically, the fermentation medium contains: 15-20 g of sodium bicarbonate, 3-5 g of sodium nitrate, 0.9-1.2 g of monopotassium phosphate and pH 7.0-7.2.
Specifically, the trace element solution contains 54g of zinc chloride, 192g of cobalt chloride, 100g of manganese sulfate, 24 g of nickel chloride, 28g of copper chloride, 38g of sodium molybdate and 30g of boric acid in 1000g of water, and has a pH value of 7.0.
The low-temperature denitrification microbial inoculum is applied to denitrification of the artificial wetland, the denitrification microbial inoculum contains nutrient solution and a preservation auxiliary agent, the denitrification microbial inoculum comprises a low-temperature nitrifying flora and denitrifying bacteria, the denitrifying bacteria are added to a water inlet area according to the proportion of the low-temperature nitrifying flora (15%) to the low-temperature aerobic denitrifying flora (10%), and the microbial inoculum is uniformly mixed in the artificial wet soil by adopting a method of uniformly adding and combining soil turning.
Specifically, the wetland water quality is characterized in that the ammonia nitrogen concentration is 30-600 mg/L, the COD concentration is 300-3000 mg/L, the BOD concentration is 200-2000 mg/L, and the pH value is 6-10.
Specifically, the denitrification fungicide is added when the concentration of the activated sludge in the wetland is 2000-5000 mg/L.
The invention has the beneficial effects that:
the invention enriches and domesticates the low-temperature nitrifying flora and the low-temperature aerobic denitrifying flora, has the best nitrifying efficiency, has the ammonia nitrogen removal rate of 85.62 percent, and can be used in the field of artificial wetland degradation treatment in winter.
Detailed Description
In the embodiment, the denitrifying bacteria are Bacillus (Bacillus) belonging to the class Bacillaceae of the phylum firmicutes, and can simultaneously carry out nitrification and denitrification processes in an aerobic or anoxic environment, and have strong tolerance to dissolved oxygen; denitrifying bacteria employ Pseudomonas belonging to the class of the Gamma-Proteobacteria of the phylum Proteobacteria (Pseudomonas).
The invention comprises the following steps:
(1) enrichment and domestication of low-temperature nitrifying bacteria: performing low-temperature acclimation on a nitrobacteria culture medium according to a temperature gradient of 30 ℃ → 25 ℃ → 20 → 15 ℃ → 13 → 10 ℃, and culturing and enriching in a mixed solution containing trace elements and nitrate and nitrogen in a 250mL shake flask under aerobic conditions, wherein the dissolved oxygen is 0.1-5 mg/L, the pH is 7.0-9.0, the shaking table rotating speed is 150rpm, the period is 12h per week, and the water discharge rate is about 90%;
(2) enrichment and domestication of low-temperature aerobic denitrification flora: activating aerobic denitrifying bacteria at the normal temperature at 30 ℃, and performing low-temperature acclimation on the activated aerobic denitrifying bacteria under the aerobic condition according to the temperature gradient of 30 ℃→ 20 → 15 ℃ → 10 ℃;
(3) inoculating the low-temperature nitrifying bacteria and the denitrifying bacteria into a fermentation culture medium for amplification culture to obtain the low-temperature aerobic denitrifying bacteria agent.
The fermentation medium contains: 15-20 g of sodium bicarbonate, 3-5 g of sodium nitrate, 0.9-1.2 g of monopotassium phosphate and pH 7.0-7.2.
The trace element solution contains 54g of zinc chloride, 192g of cobalt chloride, 100g of manganese sulfate, 24 g of nickel chloride, 28g of copper chloride, 38g of sodium molybdate and 30g of boric acid in 1000g of water, and the pH value of the solution is 7.0.
The low-temperature denitrification microbial inoculum is applied to denitrification of the artificial wetland, the denitrification microbial inoculum contains nutrient solution and a preservation auxiliary agent, the denitrification microbial inoculum comprises a low-temperature nitrifying flora and denitrifying bacteria, the denitrifying bacteria are added to a water inlet area according to the proportion of the low-temperature nitrifying flora (15%) to the low-temperature aerobic denitrifying flora (10%), and the microbial inoculum is uniformly mixed in the artificial wet soil by adopting a method of uniformly adding and combining soil turning.
The ammonia nitrogen of the outlet water of the strengthening system does not obviously decrease when the denitrifying bacteria are added, probably because the low-temperature nitrifying bacteria are required to adapt to a new environment, the transformation effect of the low-temperature nitrifying bacteria on the ammonia nitrogen is slow, then the low-temperature nitrifying bacteria start to transform the ammonia nitrogen in the inlet water, compared with a contrast system, the strengthening system has lower ammonia nitrogen concentration of the outlet water, the ammonia nitrogen concentration of the outlet water of the contrast system is about 10.73mg/L, the ammonia nitrogen concentration of the outlet water of the strengthening system is about 7.96mg/L, the first-level A standard of ammonia nitrogen emission at low temperature can be basically achieved, the ammonia nitrogen removal rates of the contrast system and the strengthening system reach about 32.09% and about 48.41%, and the removal rate of the ammonia nitrogen of the outlet water is improved by 16.32% through the. From 13d, the concentration of the ammonia nitrogen in the effluent of the strengthening system begins to rise, probably because the added microbial inoculum cannot become the dominant bacteria in the constructed wetland system, and when the biological strengthening is carried out, the repeated addition of the denitrifying bacteria by taking 12d as the addition period can be considered.
The wetland water quality is characterized in that the ammonia nitrogen concentration is 30-600 mg/L, the COD concentration is 300-3000 mg/L, the BOD concentration is 200-2000 mg/L, and the pH value is 6-10.
The denitrification fungicide is added when the concentration of the activated sludge in the wetland is 2000-5000 mg/L. In a low-temperature chamber with the temperature capable of being controlled to be 10 ℃, two parallel artificial wetlands are started to operate, artificial water distribution is used as inflow water, continuous flow operation is carried out in a horizontal subsurface flow mode under the condition that the hydraulic retention time is 24 hours until the outflow water is stable, and the water temperature is 9-10 ℃ during operation. The constructed wetland simulation system reaches a stable water outlet state when running for about 60 days at 10 ℃. In the initial stage of operation, the microbial community in the artificial wetland system is in the adaptation period, the effluent COD and ammonia nitrogen are still high, along with the operation of the system, the microorganisms in the wetland play a role, the effluent COD and ammonia nitrogen continuously decrease, after the system is operated for 40 days, the effluent quality reaches a relatively stable state, the removal rate of the effluent COD is maintained at about 47.30%, the removal rate of the effluent ammonia nitrogen is maintained at about 31.99%, the low-temperature start of the artificial wetland is completed, but the pollutant removal capacity of the artificial wetland system at low temperature is low.
As shown in Table 1, the effluent COD of the artificial wetland system is about 34.53mg/L, the effluent COD reaches the first-class A standard (50mg/L) of effluent COD discharge, the effluent ammonia nitrogen concentration is about 10.82mg/L, the effluent ammonia nitrogen discharge first-class A standard (8mg/L) at low temperature cannot be reached, the effluent nitrate nitrogen concentration is about 3.98 mg/L, the effluent total nitrogen concentration is 13.61 mg/L-15.25 mg/L, and the effluent total nitrogen discharge first-class A standard (15mg/L) cannot be stably reached, so the research performs biological enhancement of low-temperature denitrifying bacteria, reduces the effluent ammonia nitrogen concentration and the total nitrogen concentration of the artificial wetland, and enables the effluent ammonia nitrogen concentration and the total nitrogen concentration to reach the first-class A standard of the state.
TABLE 1
Figure RE-GDA0002278497440000051
The COD concentration of the effluent of the denitrifier reinforced and unreinforced systems is about 35mg/L, and the removal rate of the COD is about 46.97 percent. The COD concentration of the effluent of the control system and the enhanced system is not obviously different, probably because the added denitrifying bacteria have autotrophic bacteria and do not degrade COD, although the added heterotrophic low-temperature aerobic denitrifying bacteria in the enhanced system consume COD to a certain extent, autolyzed bacteria or metabolites of microorganisms possibly exist in the effluent, so that the COD concentration of the effluent is not obviously reduced, and sometimes is slightly higher than that of the control system.
The above description is not limited to the above examples, and the undescribed technical features of the present invention can be implemented by or using the prior art, which is not described herein again; the above examples are only for illustrating the technical solutions of the present invention and are not to be construed as limiting the present invention, the present invention is described in detail with reference to the preferred embodiments, and any modifications, equivalent substitutions, improvements, etc. made by those skilled in the art within the spirit scope of the present invention shall also fall within the protection scope of the claims of the present invention.

Claims (6)

1. A preparation method of a low-temperature denitrification microbial inoculum is characterized by comprising the following steps:
the method comprises the following steps:
(1) enrichment and domestication of low-temperature nitrifying bacteria: performing low-temperature acclimation on a nitrobacteria culture medium according to a temperature gradient of 30 ℃ → 25 ℃ → 20 → 15 ℃ → 13 → 10 ℃, and culturing and enriching in a mixed solution containing trace elements and nitrate and nitrogen in a 250mL shake flask under aerobic conditions, wherein the dissolved oxygen is 0.1-5 mg/L, the pH is 7.0-9.0, the shaking table rotating speed is 150rpm, the period is 12h per week, and the water discharge rate is about 90%;
(2) enrichment and domestication of low-temperature aerobic denitrification flora: activating aerobic denitrifying bacteria at the normal temperature at 30 ℃, and performing low-temperature acclimation on the activated aerobic denitrifying bacteria under the aerobic condition according to the temperature gradient of 30 ℃→ 20 → 15 ℃ → 10 ℃;
(3) inoculating the low-temperature nitrifying bacteria and the denitrifying bacteria into a fermentation culture medium for amplification culture to obtain the low-temperature aerobic denitrifying bacteria agent.
2. The method for preparing a low-temperature denitrification microbial inoculum according to claim 1, which is characterized in that: the fermentation medium contains: 15-20 g of sodium bicarbonate, 3-5 g of sodium nitrate, 0.9-1.2 g of monopotassium phosphate and pH 7.0-7.2.
3. The method for preparing a low-temperature denitrification microbial inoculum according to claim 1, which is characterized in that: the trace element solution contains 54g of zinc chloride, 192g of cobalt chloride, 100g of manganese sulfate, 24 g of nickel chloride, 28g of copper chloride, 38g of sodium molybdate and 30g of boric acid in 1000g of water, and the pH value of the solution is 7.0.
4. A method for denitrifying a low-temperature denitrifying microbial inoculum according to any one of claims 1 to 3 in constructed wetlands, which is characterized by comprising the following steps: the denitrification microbial inoculum contains nutrient solution and a preservation auxiliary agent, comprises a low-temperature nitrification flora and denitrification bacteria, is added into a water inlet area according to the proportion of the low-temperature nitrification flora (15%) to the low-temperature aerobic denitrification flora (10%), and is uniformly mixed in the artificial wet soil by adopting a method of uniformly adding and combining soil turning.
5. The method for denitrifying the microbial inoculum by using the low-temperature denitrifying microbial inoculum according to claim 4, which is characterized in that: the wetland water quality is characterized in that the ammonia nitrogen concentration is 30-600 mg/L, the COD concentration is 300-3000 mg/L, the BOD concentration is 200-2000 mg/L, and the pH value is 6-10.
6. The method for denitrifying the microbial inoculum by using the low-temperature denitrifying microbial inoculum according to claim 4, which is characterized in that: the denitrification fungicide is added when the concentration of the activated sludge in the wetland is 2000-5000 mg/L.
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CN111547847A (en) * 2020-05-06 2020-08-18 南京师盛生态环境研究院有限公司 Device and method for removing total nitrogen in sewage by composite microorganism enhanced A/O (anaerobic/aerobic) process
CN114451251A (en) * 2022-03-02 2022-05-10 吉林省蔬菜花卉科学研究院 Method for preparing soilless culture substrate for vegetables

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111454859A (en) * 2020-04-07 2020-07-28 山东海景天环保科技股份公司 Method for cultivating low-temperature aerobic denitrifying bacteria
CN111547847A (en) * 2020-05-06 2020-08-18 南京师盛生态环境研究院有限公司 Device and method for removing total nitrogen in sewage by composite microorganism enhanced A/O (anaerobic/aerobic) process
CN114451251A (en) * 2022-03-02 2022-05-10 吉林省蔬菜花卉科学研究院 Method for preparing soilless culture substrate for vegetables

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