CN115125164B - Highly salt-resistant and heavy metal-resistant heterotrophic nitrification-aerobic denitrification self-flocculation marine bacterium and application thereof in community construction - Google Patents
Highly salt-resistant and heavy metal-resistant heterotrophic nitrification-aerobic denitrification self-flocculation marine bacterium and application thereof in community construction Download PDFInfo
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- CN115125164B CN115125164B CN202210562929.8A CN202210562929A CN115125164B CN 115125164 B CN115125164 B CN 115125164B CN 202210562929 A CN202210562929 A CN 202210562929A CN 115125164 B CN115125164 B CN 115125164B
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- heterotrophic nitrification
- salt
- aerobic denitrification
- pseudomonas
- aerobic
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- 229910001385 heavy metal Inorganic materials 0.000 title claims abstract description 22
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- 241001288557 Pseudomonas kunmingensis Species 0.000 claims abstract description 10
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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
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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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
- C02F3/30—Aerobic and anaerobic processes
- C02F3/302—Nitrification and denitrification treatment
-
- 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
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
-
- 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
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
- C02F3/341—Consortia of bacteria
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
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- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/80—Soil conditioners
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- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/14—Soil-conditioning materials or soil-stabilising materials containing organic compounds only
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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
- C12N1/20—Bacteria; Culture media therefor
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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
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
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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
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/20—Heavy metals or heavy metal compounds
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- 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
- C02F2103/08—Seawater, e.g. for desalination
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- C12R2001/38—Pseudomonas
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/10—Biological treatment of water, waste water, or sewage
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- Medicinal Chemistry (AREA)
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- General Health & Medical Sciences (AREA)
- Soil Sciences (AREA)
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- General Life Sciences & Earth Sciences (AREA)
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Abstract
The invention provides a highly salt-resistant heavy metal-resistant heterotrophic nitrification-aerobic denitrification self-flocculation marine bacterium and application thereof in community construction, belonging to the technical field of microorganism and environmental treatment and restoration. The invention screens and obtains the pseudomonas kunmingensis (Pseudomonaskunmingens) 8-C which can utilize the heterotrophic nitrification-aerobic denitrification way to efficiently denitrify and has self-flocculation capability, has wide application value in treating high-salt wastewater, lead and cadmium polluted wastewater and soil restoration, and has the potential of developing heterotrophic nitrification-aerobic denitrification communities with good sedimentation performance. Therefore, the method has good practical application value.
Description
Technical Field
The invention belongs to the technical field of microorganism and environmental treatment and repair, and particularly relates to a highly salt-resistant and heavy metal-resistant heterotrophic nitrification-aerobic denitrification self-flocculation marine bacterium and application thereof in community construction.
Background
The disclosure of this background section is only intended to increase the understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art already known to those of ordinary skill in the art.
The exceeding of the standard of the nitrogen-containing compounds in the water body can cause eutrophication of the water body, cause algae and other plankton to rapidly reproduce, reduce the dissolved oxygen of the water body, deteriorate the water quality, kill a large amount of fishes and other organisms, and seriously endanger ecological balance and human health. The biological denitrification method is widely applied to the treatment of wastewater nitrogen pollution due to the characteristics of economy, high efficiency, no secondary pollution and the like. The conventional biological denitrification technology needs to go through two steps, namely an aerobic nitrification process of autotrophic nitrifying bacteria and an anaerobic denitrification process of abnormal denitrifying bacteria. The anoxic-aerobic process is the most basic engineering example of the traditional biological denitrification technology. In the anoxic section, denitrifying bacteria utilize organic carbon in sewage as an electron donor and nitrate as an electron acceptor to perform anaerobic respiration, so that nitrate nitrogen in the reflux liquid is reduced into nitrogen gas to be released, and the denitrification process is completed. In the aerobic section, nitrifying bacteria oxidize ammonia nitrogen in sewage into nitrate and then flow back to the anoxic tank. The technology requires strict control of aerobic and anaerobic conditions in two stages and therefore must be run in two separate structures. And nitrifying bacteria have long generation time and require longer sludge retention time. Bacteria performing the two functions have a large difference in growth conditions, metabolic characteristics and the like, thereby increasing investment cost and complexity of regulation in practical application. Finding a more economical, faster, and more efficient process is therefore an important concern for biological denitrification.
In recent years, heterotrophic nitrification-aerobic denitrification bacteria have been found from the environment by researchers, and such microorganisms are capable of simultaneously performing nitrification and denitrification under aerobic conditions. The discovery of the process breaks through the bottleneck of biological denitrification technology. However, the inventor finds that most heterotrophic nitrification-aerobic denitrification strains screened at present need to grow in a mild and proper environment, have high requirements on environmental conditions, cannot be efficiently denitrified in high-salt heavy metal wastewater, have poor sedimentation performance, are not suitable for practical engineering application, and are limited to single bacteria, so that a microorganism still capable of efficiently degrading nitrogen pollutants in extreme environments such as saline alkali, heavy metals and the like needs to be searched, and the potential of the microorganism in the aspects of community synthesis, microbiome construction and the like is mined.
Disclosure of Invention
Based on the defects of the prior art, the invention provides a highly salt-resistant heavy metal-resistant heterotrophic nitrification-aerobic denitrification self-flocculation marine bacterium and application thereof in community construction. The invention screens and obtains the pseudomonas kunmingensis (Pseudomonas kunmingens) 8-C which can utilize the heterotrophic nitrification-aerobic denitrification way to efficiently denitrify and has self-flocculation capability, has wide application value in treating high-salt wastewater, lead and cadmium polluted wastewater and soil restoration, and has the potential of developing heterotrophic nitrification-aerobic denitrification communities with good sedimentation performance. Based on the above results, the present invention has been completed.
In order to achieve the technical purpose, the invention relates to the following technical scheme:
In one aspect of the present invention, there is provided a heterotrophic nitrification-aerobic denitrifying bacteria, classified as Pseudomonas kumi (Pseudomonas kunmingens) 8-C, which has been deposited with the China center for type culture Collection on 12 months 13 of 2021 (address: university of Wuchang Lojia mountain, wuhan, hubei province), with a biological deposit number of CCTCC NO: m20211592.
In a second aspect of the present invention, there is provided a microbial agent comprising the heterotrophic nitrification-aerobic denitrification bacterium 8-C described above.
In a third aspect of the present invention, there is provided a salt-tolerant heterotrophic nitrification-aerobic denitrification microbiome comprising the heterotrophic nitrification-aerobic denitrification bacterium 8-C; more specifically, the salt-tolerant heterotrophic nitrification-aerobic denitrification microbiome is obtained by inoculating the heterotrophic nitrification-aerobic denitrification bacterium 8-C and acinetobacter johnsonii (Acinetobacter johnsonii) 2-1-H into high-salt wastewater for culture; the salt-adapting heterotrophic nitrification-aerobic denitrification microbiome composed of the heterotrophic nitrification-aerobic denitrification bacteria 8-C and the acinetobacter johnsonii 2-1-H has good and stable treatment effect, the ammonia nitrogen removal rate in the simulated high-salt wastewater can reach 90%, the total nitrogen removal rate can reach 85%, and the accumulation of nitrite and nitrate is almost avoided.
Wherein, the Acinetobacter johnsonii (Acinetobacter johnsonii) 2-1-H is preserved in China center for type culture collection (address: university of Wuchang mountain and Wuhan in Wuhan, hubei province) on 12 th month 13 days of 2021, and the biological preservation number is CCTCC NO: m20211593.
In a fourth aspect of the present invention, there is provided a method for constructing the above-described salt-tolerant heterotrophic nitrification-aerobic denitrification microbiome, the method comprising:
Inoculating the heterotrophic nitrification-aerobic denitrification bacteria 8-C and acinetobacter johnsonii 2-1-H into a bioreactor containing high-salt wastewater, and operating until the heterotrophic nitrification-aerobic denitrification bacteria have stable ammonia nitrogen removal efficiency and sedimentation performance, thus obtaining the salt-suitable heterotrophic nitrification-aerobic denitrification microbiome.
The reactor is not subjected to sludge discharge in operation, so that the salt-tolerant heterotrophic nitrification-aerobic denitrification microbiome is easier to obtain.
In a fifth aspect of the invention, the use of the heterotrophic nitrification-aerobic denitrification bacteria, microbial agents and/or salt-tolerant heterotrophic nitrification-aerobic denitrification microbiome in any one or more of the following:
a) Waste water treatment;
b) Preparing an organic fertilizer;
c) Soil improvement;
d) Treating water eutrophication;
e) Repairing water pollution;
f) Biological denitrification.
In a sixth aspect of the present invention, there is provided a method for integrated, resource conversion of nutrients in a high salt environment, the method comprising: the heterotrophic nitrification-aerobic denitrification microorganisms and/or the halophilic heterotrophic nitrification-aerobic denitrification microbiome are applied to a high-salt environment.
More specifically, the high-salt environment is a high-salt water environment, and further can be a high-salt water environment containing heavy metal pollution; the salinity of the high-salinity water environment is not lower than 3%, and is more preferably 3% -8%; the heavy metals include, but are not limited to, lead and cadmium.
The beneficial technical effects of one or more of the technical schemes are as follows:
According to the technical scheme, the pseudomonas kunmingensis (Pseudomonas kunmingens) 8-C which can be efficiently denitrified by utilizing a heterotrophic nitrification-aerobic denitrification way and has self-flocculation capacity is obtained through screening and separation, and researches show that the pseudomonas kunmingensis can utilize ammonia nitrogen and nitrate as nitrogen sources under an aerobic condition, no nitrite nitrogen is accumulated in the ammonia nitrogen conversion process, a small amount of nitrate is produced and then removed, and the pseudomonas kunmingensis has wide tolerance range to salinity and pH and good tolerance to heavy metals such as lead and cadmium and the like; meanwhile, further researches show that the heterotrophic nitrification-aerobic denitrification microbiome obtained by synthesizing the same with Acinetobacter johnsonii (Acinetobacter johnsonii) 2-1-H has good sedimentation performance and stable and efficient denitrification capability, and the total nitrogen removal rate can reach 90% in simulated high-salt wastewater, and the total nitrogen removal rate can reach 85%, and the accumulation of nitrite and nitrate is almost avoided, so that the method has wide industrial application prospect in biological denitrification pollution treatment, particularly in treatment of high-salt and heavy metal polluted ammonia nitrogen wastewater.
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention.
FIG. 1 is a phylogenetic tree of the invention of the salt-tolerant autotrophic nitrification-aerobic denitrification bacterium Pseudomonas kumi (Pseudomonas kunmingens) 8-C;
FIG. 2 shows the macroscopic and microscopic surface morphology of Pseudomonas qunmingensis (Pseudomonas kunmingens) 8-C of the present invention;
FIG. 3 shows the effect of Pseudomonas qunmingensis (Pseudomonas kunmingens) 8-C on ammonia nitrogen and nitrate nitrogen removal in the present invention;
FIG. 4 is a graph showing the total nitrogen balance of Pseudomonas qunmingensis (Pseudomonas kunmingens) 8-C of the present invention using ammonia nitrogen and nitrate;
FIG. 5 shows the ammonia nitrogen removal effect of Pseudomonas qunmingensis (Pseudomonas kunmingens) 8-C of the present invention under different salinity conditions;
FIG. 6 shows the ammonia nitrogen removal effect of Pseudomonas qunmingensis (Pseudomonas kunmingens) 8-C of the present invention at various pH conditions;
FIG. 7 shows the ammonia nitrogen removal effect of Pseudomonas qunmingensis (Pseudomonas kunmingens-C) in simulated wastewater containing Pb 2+、Cd2+ in accordance with the present invention.
FIG. 8 shows the use of two strains of salt-tolerant HNAD bacteria: pseudomonas kumi (Pseudomonas kunmingens) 8-C, acinetobacter johnsonii (Acinetobacter johnsonii) 2-1-H constructed colonies.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the invention. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments in accordance with the present application. As used herein, the singular is also intended to include the plural unless the context clearly indicates otherwise, and furthermore, it is to be understood that the terms "comprises" and/or "comprising" when used in this specification are taken to specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof.
As described above, most heterotrophic nitrification-aerobic denitrification strains screened at present need to grow in a moderate and proper environment, have high requirements on environmental conditions, cannot be efficiently denitrified in high-salt heavy metal wastewater, have poor sedimentation performance, are not suitable for practical engineering application, and are currently mostly limited to single bacteria, so that a microorganism still capable of efficiently degrading nitrogen pollutants in extreme environments such as saline alkali, heavy metals and the like needs to be searched, and the potential of the microorganism in aspects of community synthesis, microbiome construction and the like is mined.
Accordingly, in one embodiment of the present invention, a heterotrophic nitrification-aerobic denitrifying bacteria, classified as Pseudomonas kunmingensis (Pseudomonas kunmingens) 8-C, which has been deposited at the China center for type culture collection (address: university of Wuchang Lojia mountain, wuhan, hubei province) under the biological deposit number CCTCC NO: m20211592.
The thallus and colony characteristics are: the strain is gram negative bacteria, the bacteria are rod-shaped when observed by a scanning electron microscope, the bacterial body size is about 0.5-1.0 mu m, and bacterial colonies on an LB solid medium are milky white, moist, smooth and flat.
According to the invention, researches show that the pseudomonas kunmingensis 8-C can utilize ammonia nitrogen and nitrate as nitrogen sources under an aerobic condition, no nitrite nitrogen is accumulated in the ammonia nitrogen conversion process, and a small amount of nitrate is produced and then removed. The total nitrogen in the system is converted from the water phase to the gas phase and into biomass in the process of utilizing ammonia nitrogen and nitrate, and about 50% of nitrogen is lost in a gas state.
Meanwhile, the strain has higher tolerance to salinity conditions. Compared with low salinity, the strain has higher ammonia nitrogen removal effect under medium and high salinity. The ammonia nitrogen removal rate exceeds 85% in the salinity range of 3-8%; when the salinity is 1-2%, the ammonia nitrogen removal rate is also more than 40%. The strain has wide tolerance range to pH, is more favorable for the growth and metabolism under neutral and weak alkaline conditions, and has ammonia nitrogen removal efficiency of more than 90 percent in the pH range of 7-9.
In addition, the strain has good tolerance to Pb 2+、Cd2+ and other heavy metal ions, the ammonia nitrogen removal rate can reach 85% under the stress of 20mg/L Pb 2+, and the ammonia nitrogen removal rate can reach 80% under the stress of 10mg/L Cd 2+. Thereby further widening the application field and application range thereof.
In still another embodiment of the present invention, there is provided a microbial agent comprising the heterotrophic nitrification-aerobic denitrification bacterium 8-C described above.
In still another embodiment of the present invention, the microbial agent contains a carrier in addition to the heterotrophic nitrification-aerobic denitrification bacteria 8-C as an active ingredient. The carrier may be a carrier commonly used in the field of bacteriological agents and which is biologically inert.
The carrier may be a solid carrier or a liquid carrier;
The solid carrier can be mineral material, plant material or high molecular compound; the mineral material may be at least one of clay, talc, kaolin, montmorillonite, white carbon, zeolite, silica, and diatomaceous earth; the plant material may be at least one of corn flour, soy flour and starch; the high molecular compound can be polyvinyl alcohol or/and polyglycol;
The liquid carrier may be an organic solvent, vegetable oil, mineral oil, or water; the organic solvent can be decane or/and dodecane.
The dosage form of the microbial inoculum can be various dosage forms, such as liquid, emulsion, suspending agent, powder, granule, wettable powder or water dispersible granule.
Surfactants (such as Tween 20, tween 80, etc.), binders, stabilizers (such as antioxidants), pH regulators, etc. can also be added into the microbial inoculum according to the need.
In still another embodiment of the present invention, there is provided an adaptive salt heterotrophic nitrification-aerobic denitrification microbiome comprising the aforementioned heterotrophic nitrification-aerobic denitrification bacterium 8-C; more specifically, the salt-tolerant heterotrophic nitrification-aerobic denitrification microbiome is obtained by inoculating the heterotrophic nitrification-aerobic denitrification bacterium 8-C and acinetobacter johnsonii (Acinetobacter johnsonii) 2-1-H into high-salt wastewater for culture; the salt-adapting heterotrophic nitrification-aerobic denitrification microbiome composed of the heterotrophic nitrification-aerobic denitrification bacteria 8-C and the acinetobacter johnsonii 2-1-H has good and stable treatment effect, the ammonia nitrogen removal rate in the simulated high-salt wastewater can reach 90%, the total nitrogen removal rate can reach 85%, and the accumulation of nitrite and nitrate is almost avoided.
Wherein, the Acinetobacter johnsonii (Acinetobacter johnsonii) 2-1-H is preserved in China center for type culture collection (address: university of Wuchang mountain and Wuhan in Wuhan, hubei province) on 12 th month 13 days of 2021, and the biological preservation number is CCTCC NO: m20211593.
In still another embodiment of the present invention, the salt-tolerant nitrogen assimilating microbiome may be high-salt wastewater obtained by the above-described treatment or activated sludge obtained by the above-described treatment.
In still another embodiment of the present invention, there is provided a method for constructing the above-described halophilic nitrogen-assimilating microbiome, comprising:
Inoculating the heterotrophic nitrification-aerobic denitrification bacteria 8-C and acinetobacter johnsonii 2-1-H into a bioreactor containing high-salt wastewater, and operating until the heterotrophic nitrification-aerobic denitrification bacteria have stable ammonia nitrogen removal efficiency and sedimentation performance, thus obtaining the salt-suitable heterotrophic nitrification-aerobic denitrification microbiome.
The reactor is not subjected to sludge discharge in operation, so that the salt-tolerant heterotrophic nitrification-aerobic denitrification microbiome is easier to obtain.
The high-salt wastewater can be actual high-salt wastewater or simulated high-salt wastewater; wherein, the simulated high-salt wastewater can be seawater simulated wastewater, and the specific components comprise: 3% of seawater element, 500mg/L of sodium acetate, 140mg/L of dipotassium hydrogen phosphate and 550mg/L of ammonium chloride.
The bioreactor is a sequencing batch bioreactor (SBR), and the sequencing batch reactor is an intermittent activated sludge system adopting a tank body which is used as the bioreactor and a sedimentation tank. When treating continuous flow sewage, at least two or more tanks are needed, and an air diffusion device is arranged at the bottom of the tank body to play roles in aeration and stirring.
The construction method comprises the following steps:
the operation is continuous, each cycle comprises 5min water inlet, 450min aeration, 15min sedimentation and 10min water outlet, each cycle is 8h, the volume exchange rate is 62.5%, and the Hydraulic Retention Time (HRT) is 12.8h.
In yet another embodiment of the present invention, the use of the heterotrophic nitrification-aerobic denitrification bacteria, microbial agents, and/or salt-tolerant heterotrophic nitrification-aerobic denitrification microbiome as described above in any one or more of the following:
a) Waste water treatment;
b) Preparing an organic fertilizer;
c) Soil improvement;
d) Treating water eutrophication;
e) Repairing water pollution;
f) Biological denitrification.
In said a), the waste water includes, but is not limited to, high salt waste water, marine culture waste water, industrial salty sewage and seawater toilet flushing waste water; the wastewater may also be contaminated with heavy metals including, but not limited to, lead and cadmium.
In the c), the soil may be saline-alkali soil; the soil may also be a saline-alkali soil contaminated with heavy metals including, but not limited to, lead and cadmium; the soil improvement may be soil fertility improvement.
In said d), said body of water comprises fresh water and sea water, preferably sea water.
In yet another embodiment of the present invention, a method for integrated, resource-based conversion of nutrients in a high-salt environment is provided, the method comprising: the heterotrophic nitrification-aerobic denitrification microorganisms and/or the halophilic heterotrophic nitrification-aerobic denitrification microbiome are applied to a high-salt environment.
More specifically, the high-salt environment is a high-salt water environment, and further can be a high-salt water environment containing heavy metal pollution; the salinity of the high-salinity water environment is not lower than 3%, and is more preferably 3% -8%; the heavy metals include, but are not limited to, lead and cadmium.
The invention is further illustrated by the following examples, which are not to be construed as limiting the invention. It is to be understood that these examples are illustrative of the present invention and are not intended to limit the scope of the present invention. In the embodiment, the determination and analysis methods of NH 4 +-N、NO2 --N、NO3 - -N, TN are all referred to national standard, wherein the determination and analysis of NH 4 + -N are performed according to the method of spectrophotometry for determination of Water quality and Ammonia nitrogen and Nashi reagent (GB HJ 535-2009); determination and analysis of NO 2 - -N according to the Spectrophotometry of Water quality-determination of nitrite Nitrogen (GB 7493-87); determination and analysis of NO 3 - -N according to Water quality-determination of nitrate nitrogen-ultraviolet spectrophotometry (GB HJ/T346-2007); TN determination and analysis is carried out according to the "determination of Water quality-total Nitrogen-ultraviolet spectrophotometry" (GB 11894-89).
The basal medium used in the examples was autoclaved at 121℃for 20min and the formulation was as follows:
(1) Trace element solution (L-1)：EDTA50g、ZnSO4·7H2O 5.02g、CuSO4·5H2O 1.57g、FeSO4·7H2O 5.0g、CaCl2·2H2O 5.5g、MnCl2·4H2O 5.06g、pH 6.0.
(2) Enrichment medium (L-1)：MgSO4·7H2O 0.01g、KH2PO4 0.056g、Na2HPO4 7.9g、NH4Cl 0.192g、NaNO3 0.3415g、NaNO2 0.362g、CH3COONa 0.65g、 seawater 30g and trace element solution 2mL.
(3) Heterotrophic Nitrification (HNM) pre-screening culture medium (L -1)：CH3COONa 0.65g、NH4Cl 0.192g、KH2PO4 0.056g, seawater 30g, trace element solution 2mL.
(4) BTB medium (L -1): 1mL of bromothymol blue ethanol solution, CH 3COONa 0.65g、NaNO30.8415g、KH2PO4 0.056.056 g and agar 15g.
(5) Aerobic Denitrification (DEM) pre-screening culture medium (L -1)：CH3COONa 0.65g、NaNO3 0.3415g、KH2PO4 0.056g, seawater 30g, trace element solution 2 mL).
Example 1
Enrichment, separation, screening and identification of heterotrophic nitrification-aerobic denitrification strains
(1) Pretreatment of submarine sediment: the experimental seabed sediment is taken from yellow sea A4 (122 DEG 48'E,35 DEG 59' N), 10g of the seabed sediment is put into a 300mL wide-mouth triangular flask filled with 90mL sterile physiological saline with the concentration of 0.9% in an ultra-clean workbench, a little glass beads which are subjected to high-pressure steam sterilization at 121 ℃ for 15min are put into the triangular flask, and 180r/min oscillation is carried out for 1h to break up a sediment sample, so that microorganisms in the sediment are fully suspended in the physiological saline.
(2) Enrichment culture of strains: 20mL of the sediment pretreatment mixed solution is taken and added into a 500mL conical flask filled with 200mL of seawater LB culture medium, and the mixture is cultured for 48h at 25 ℃ and 180r/min in a shaking table.
(3) Separating and purifying strains: and respectively taking 1mL of water samples of the upper layer, the middle layer and the lower layer of the enrichment culture medium, respectively inoculating the water samples into a centrifuge tube filled with 9mL of sterile physiological saline, and repeating the steps to sequentially dilute the water samples to reach a concentration gradient of 10 -2～10-8. Then 100L of each concentration gradient mixed solution is respectively coated on a BTB solid culture medium, and the mixture is inversely cultured for 2 to 3 days in a constant temperature incubator at 25 ℃ until single colony with blue halo grows. The single colony is picked by an inoculating loop, and repeated line drawing and separation are carried out on a new LB solid medium for a plurality of times by adopting a flat plate streak separation method until a pure colony is obtained.
(4) Screening and identification of strains: the purified strains are picked up and respectively inoculated in heterotrophic nitrification pre-screening culture medium, and cultured for 48 hours in a constant temperature shaking table at 25 ℃ and 180 rpm. Taking culture solutions of 12 hours, 24 hours and 48 hours, measuring OD 600, 12000rpm and 5min, centrifuging, taking supernatant, and measuring the concentration of NH 4 +-N、NO2 --N、NO3 - -N. Selecting the microorganism with highest ammonia nitrogen removal efficiency, least nitrite and nitrate accumulation and self-flocculation capacity, namely the pseudomonas kunmingensis (Pseudomonas kunmingens) 8-C. As shown in FIG. 1, the bacterial colony morphology of Pseudomonas (Pseudomonas kunmingens) 8-C obtained in this example: the single bacterial colony is light yellow, the surface is convex and glossy, the bacterial colony is small in viscosity and not easy to pick, the edge is neat, and the diameter of the bacterial colony is smaller than that of other bacterial strains screened out, namely about 0.5mm-1.0mm; SEM shows that the pseudomonas (Pseudomonas kunmingens) 8-C is in a rod shape and slightly bent, the thallus is slender, and the single cell length is about 2.5m; the measured 16srDNA sequences were entered into the GenBank database website of the NCBI website for similarity comparison and phylogenetic trees were drawn. The phylogenetic tree results are shown in FIG. 2, and the results show that Pseudomonas (Pseudomonas kunmingens) 8-C has higher similarity with Pseudomonas sp.HSI-7 and Pseudomonaskunmingens strain P P-1-1, so the strain is named Pseudomonas kunmingens-C. The 16srDNA sequence is as follows:
GGCGTGGGCAAAAAGCTACCTGCTAGTCGAGCGGATGAAGAGAGCTTGCTTTCTGATTCAGCGGCGGACGGGTGAGTAATGCCTAGGAATCTGCCTGATAGTGGGGGACAACGTTTCGAAAGGAACGCTAATACCGCATACGTCCTACGGGAGAAAGCAGGGGACCTTCGGGCCTTGCGCTATCAGATGAGCCTAGGTCGGATTAGCTAGTTGGTGAGGTAACGGCTCACCAAGGCGACGATCCGTAACTGGTCTGAGAGGATGATCAGTCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGCGAAAGCCTGATCCAGCCATGCCGCGTGTGTGAAGAAGGTCTTCGGATTGTAAAGCACTTTAAGTTGGGAGGAAGGGCATTAACCTAATACGTTAGTGTTTTGACGTTACCGACAGAATAAGCACCGGCTAACTTCGTGCCAGCAGCCGCGGTAATACGAAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCGCGTAGGTGGTTTGTTAAGTTGAATGTGAAAGCCCCGGGCTCAACCTGGGAACTGCATCCAAAACTGGCAAGCTAGAGTATGGCAGAGGGTGGTGGAATTTCCTGTGTAGCGGTGAAATGCGTAGATATAGGAAGGAACACCAGTGGCGAAGGCGACCACCTGGGCTAATACTGACACTGAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCGACTAGCCGTTGGGATCCTTGAGATCTTAGTGGCGCAGCTAACGCATTAAGTCGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGCCTTGACATGCAGAGAACTTTCCAGAGATGGATTGGTGCCTTCGGGAACTCTGACACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGTAACGAGCGCAACCCTTGTCCTTAGTTACCAGCACGTTAAGGTGGGCACTCTAAGGAGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGGCCCTTACGGCCTGGGCTACACACGTGCTACAATGGTCGGTACAAAGGGTTGCCAAGCCGCGAGGTGGAGCTAATCCCATAAAACCGATCGTAGTCCGGATCGCAGTCTGCAACTCGACTGCGTGAAGTCGGAATCGCTAGTAATCGTGAATCAGAATGTCACGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTGCTCCAGAAGTAGCTAGTCTAACATCGGGGGGCACAGTACCACTGAGGAT(SEQ ID NO.1)
example 2
Determination of the metabolic Properties of Ammonia nitrogen and nitrate of Pseudomonas (Pseudomonas kunmingens) 8-C
Inoculating Pseudomonas (Pseudomonas kunmingens) 8-C into seawater LB culture medium, and shake culturing at 25deg.C and 180r/min for 24 hr to obtain seed solution for subsequent experiment. Centrifuging the prepared Pseudomonas (Pseudomonas kunmingens) 8-C seed solution at 4000rpm for 5min; removing supernatant, re-suspending with sterilized normal saline (0.85% seawater element) and washing the thallus; centrifuging at 4000rpm for 5min, and discarding supernatant; re-suspending the bacterial strain with sterilized physiological saline, inoculating 10% of the bacterial strain into a culture medium with ammonia nitrogen and nitrate as single nitrogen sources, and sampling according to time points to determine the utilization and conversion of the bacterial strain to various nitrogen sources.
The culture mediums used in the method are HNM culture medium and DEM culture medium.
As can be seen from FIG. 3, the strain can utilize ammonia nitrogen and nitrate as nitrogen sources under aerobic conditions, no nitrite nitrogen is accumulated in the ammonia nitrogen conversion process, and a small amount of nitrate is produced and then removed. As can be seen from fig. 4, the strain converts total nitrogen in the system from the water phase to the gas phase and into biomass in the process of utilizing ammonia nitrogen and nitrate, and about 50% of nitrogen is lost in the gas state.
Example 3
Pseudomonas (Pseudomonas kunmingens) 8-C optimal growth conditions and denitrification conditions
Inoculating Pseudomonas (Pseudomonas kunmingens) 8-C into seawater LB culture medium, and shake culturing at 25deg.C and 180r/min for 24 hr to obtain seed solution for subsequent experiment. Centrifuging the prepared Pseudomonas (Pseudomonas kunmingens-C) seed solution at 4000rpm for 5min; removing supernatant, re-suspending with sterilized normal saline (0.85% seawater element) and washing the thallus; centrifuging at 4000rpm for 5min, and discarding supernatant; re-suspending the bacterial cells with sterilized normal saline, respectively inoculating the bacterial cells into HNM culture media in an inoculum size of 10%, exploring the influence of different salinity and pH on the denitrification efficiency of the bacterial strain, and sampling and determining the utilization and conversion of the bacterial strain to ammonia nitrogen according to time points.
The culture medium used in the method is HNM culture medium, the simulated wastewater with different salinity is prepared from 1%, 2%, 3%, 5% and 8% (w/w) of seawater, and the simulated wastewater with different pH is prepared from (1+2) M HCl and (1+4) M NaOH respectively.
As can be seen from FIG. 5, the strain has a high tolerance to salinity conditions. Compared with low salinity, the strain has higher ammonia nitrogen removal effect under medium and high salinity. The ammonia nitrogen removal rate exceeds 85% in the salinity range of 3-8%; when the salinity is 1-2%, the ammonia nitrogen removal rate is also more than 40%.
As can be seen from FIG. 6, the strain has wide pH tolerance range, and neutral and weak alkaline conditions are more favorable for growth and metabolism, and the ammonia nitrogen removal efficiency exceeds 90% in the pH range of 7-9.
Example 4
Pseudomonas (Pseudomonas kunmingens) 8-C removes ammonia nitrogen under Pb 2+、Cd2+ stress
Inoculating Pseudomonas (Pseudomonas kunmingens) 8-C into seawater LB culture medium, and shake culturing at 25deg.C and 180r/min for 24 hr to obtain seed solution for subsequent experiment. Centrifuging the prepared Pseudomonas (Pseudomonas kunmingens) 8-C seed solution at 4000rpm for 5min; removing supernatant, re-suspending with sterilized normal saline (0.85% seawater element) and washing the thallus; centrifuging at 4000rpm for 5min, and discarding supernatant; re-suspending the thallus with sterilized physiological saline, inoculating into HNM culture medium in 10% amount, and measuring the denitrification capacity of the strain in Pb 2+、Cd2+ containing waste water.
As can be seen from FIG. 7, the Pseudomonas (Pseudomonas kunmingens-C) has better tolerance to Pb 2+、Cd2+, the ammonia nitrogen removal rate can reach 85% under 20mg/L of Pb 2+ stress, and the ammonia nitrogen removal rate can reach 80% under 10mg/L of Cd 2+ stress.
Example 5
Pseudomonas (Pseudomonas kunmingens) 8-C and Acinetobacter (Acinetobacter johnsonii) 2-1-H construct communities
A method for developing a salt-adapting heterotrophic nitrification-aerobic denitrification microbiome by using pseudomonas (Pseudomonas kunmingens) 8-C and another strain HNAD acinetobacter (Acinetobacter johnsonii) 2-1-H comprises the following steps:
(1) Respectively inoculating Pseudomonas (Pseudomonas kunmingens-C) and Acinetobacter (Acinetobacter johnsonii-1-H) into seawater LB culture medium, and shake culturing at 25deg.C and 180r/min for 24 hr to obtain activated bacterial liquid;
(2) Mixing the pseudomonas (Pseudomonas kunmingens) 8-C and the acinetobacter (Acinetobacter johnsonii) 2-1-H bacterial bodies prepared in the step (1) according to a volume ratio of 1:1 in a sequencing batch bioreactor (SBR), aeration for 24 hours followed by sedimentation for 12 hours, discarding the supernatant, adding an equal volume of simulated wastewater, and repeating this step until the LB is completely replaced. The effective volume of the sequencing batch bioreactor (SBR) is 3.2L, and an air diffusion device is arranged at the bottom of the sequencing batch bioreactor to play a role in aeration and stirring. The reactor was operated in a continuous mode with each cycle comprising 5min inlet, 450min aeration, 15min sedimentation and 10min outlet, 8h per cycle, 62.5% volume exchange and 12.8h Hydraulic Retention Time (HRT). The sequencing batch bioreactor operating parameters are shown in table 1.
TABLE 1
The simulated seawater waste water components in the step (2) are shown in table 2:
TABLE 2
The reactor is not discharged during operation, so that a suitable salt heterotrophic nitrification-aerobic denitrification microbial community with stable ammonia nitrogen removal efficiency and good sedimentation performance is obtained, and the total nitrogen, ammonia nitrogen, nitrite and nitrate concentration of the suitable salt heterotrophic nitrification-aerobic denitrification microbial community is measured by sampling at regular time. As shown in the running result in figure 8, the result shows that the heterotrophic nitrification-aerobic denitrification microbiome obtained by synthesizing the community by using the pseudomonas (Pseudomonas kunmingens) 8-C and the acinetobacter (Acinetobacter johnsonii) 2-1-H and developing has good and stable treatment effect, the ammonia nitrogen removal rate in the simulated high-salt wastewater can reach 90%, the total nitrogen removal rate can reach 85%, and the accumulation of nitrite and nitrate is almost avoided. The reactor continuously runs for 40 days, the biomass growth rate is low, the sludge yield is low, the sludge discharge frequency is reduced, the energy and resource consumption in the subsequent sludge reduction treatment is reduced, and the method is provided for the treatment of the heavy metal-containing high-salinity wastewater.
Finally, it should be noted that the above-mentioned embodiments are only preferred embodiments of the present invention, and the present invention is not limited to the above-mentioned embodiments, but may be modified or substituted for some of them by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention. While the foregoing describes the embodiments of the present invention, it should be understood that the present invention is not limited to the embodiments, and that various modifications and changes can be made by those skilled in the art without any inventive effort.
SEQUENCE LISTING
<110> University of Shandong
<120> A highly salt-tolerant, heavy metal-tolerant heterotrophic nitrification-aerobic denitrification self-flocculating marine bacterium and its use in community structure
Application in construction
<130>
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1435
<212> DNA
<213> Pseudomonas kunmingensis Pseudomonas kunmingens
<400> 1
ggcgtgggca aaaagctacc tgctagtcga gcggatgaag agagcttgct ttctgattca 60
gcggcggacg ggtgagtaat gcctaggaat ctgcctgata gtgggggaca acgtttcgaa 120
aggaacgcta ataccgcata cgtcctacgg gagaaagcag gggaccttcg ggccttgcgc 180
tatcagatga gcctaggtcg gattagctag ttggtgaggt aacggctcac caaggcgacg 240
atccgtaact ggtctgagag gatgatcagt cacactggaa ctgagacacg gtccagactc 300
ctacgggagg cagcagtggg gaatattgga caatgggcga aagcctgatc cagccatgcc 360
gcgtgtgtga agaaggtctt cggattgtaa agcactttaa gttgggagga agggcattaa 420
cctaatacgt tagtgttttg acgttaccga cagaataagc accggctaac ttcgtgccag 480
cagccgcggt aatacgaagg gtgcaagcgt taatcggaat tactgggcgt aaagcgcgcg 540
taggtggttt gttaagttga atgtgaaagc cccgggctca acctgggaac tgcatccaaa 600
actggcaagc tagagtatgg cagagggtgg tggaatttcc tgtgtagcgg tgaaatgcgt 660
agatatagga aggaacacca gtggcgaagg cgaccacctg ggctaatact gacactgagg 720
tgcgaaagcg tggggagcaa acaggattag ataccctggt agtccacgcc gtaaacgatg 780
tcgactagcc gttgggatcc ttgagatctt agtggcgcag ctaacgcatt aagtcgaccg 840
cctggggagt acggccgcaa ggttaaaact caaatgaatt gacgggggcc cgcacaagcg 900
gtggagcatg tggtttaatt cgaagcaacg cgaagaacct taccaggcct tgacatgcag 960
agaactttcc agagatggat tggtgccttc gggaactctg acacaggtgc tgcatggctg 1020
tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgta acgagcgcaa cccttgtcct 1080
tagttaccag cacgttaagg tgggcactct aaggagactg ccggtgacaa accggaggaa 1140
ggtggggatg acgtcaagtc atcatggccc ttacggcctg ggctacacac gtgctacaat 1200
ggtcggtaca aagggttgcc aagccgcgag gtggagctaa tcccataaaa ccgatcgtag 1260
tccggatcgc agtctgcaac tcgactgcgt gaagtcggaa tcgctagtaa tcgtgaatca 1320
gaatgtcacg gtgaatacgt tcccgggcct tgtacacacc gcccgtcaca ccatgggagt 1380
gggttgctcc agaagtagct agtctaacat cggggggcac agtaccactg aggat 1435
Claims (14)
1. A heterotrophic nitrification-aerobic denitrifying bacterium, which is characterized by being classified and named as pseudomonas kunmingensis (Pseudomonas kunmingens) 8-C, and being preserved in China Center for Type Culture Collection (CCTCC) at 12-13 of 2021, wherein the biological preservation number is CCTCC NO: m20211592.
2. A microbial agent comprising the heterotrophic nitrification-aerobic denitrification bacterium of claim 1.
3. The microbial agent of claim 2, further comprising a carrier.
4. A microbial agent according to claim 3, wherein the carrier is a solid carrier or a liquid carrier.
5. An adaptive heterotrophic nitrification-aerobic denitrification microbiome comprising the heterotrophic nitrification-aerobic denitrification bacterium of claim 1.
6. The salt-tolerant heterotrophic nitrification-aerobic denitrification microbiome of claim 5, wherein said salt-tolerant heterotrophic nitrification-aerobic denitrification microbiome is obtained by inoculating said heterotrophic nitrification-aerobic denitrification bacteria and acinetobacter johnsonii (Acinetobacterjohnsonii) 2-1-H into high-salt wastewater for cultivation;
Wherein, the acinetobacter johnsonii (Acinetobacterjohnsonii) 2-1-H is preserved in China center for type culture collection (China center for type culture collection) on 12 months 13 of 2021, and the biological preservation number is CCTCC NO: m20211593.
7. The method for constructing the salt-tolerant heterotrophic nitrification-aerobic denitrification microbiome as set forth in claim 6, characterized in that said method of constructing is as follows:
Inoculating the heterotrophic nitrification-aerobic denitrification bacteria and acinetobacter johnsonii 2-1-H into a bioreactor containing high-salt wastewater, and operating until the heterotrophic nitrification-aerobic denitrification bacteria and acinetobacter johnsonii have stable ammonia nitrogen removal efficiency and sedimentation performance, thereby obtaining the salt-suitable heterotrophic nitrification-aerobic denitrification microbiome.
8. The method of claim 7, wherein the bioreactor is a sequencing batch bioreactor.
9. The construction method according to claim 8, wherein the construction method comprises:
the operation is continuous, each cycle comprises 5min water inflow, 450min aeration, 15min sedimentation and 10min water outflow, each cycle is 8h, the volume exchange rate is 62.5%, and the hydraulic retention time is 12.8h.
10. Use of the heterotrophic nitrification-aerobic denitrification bacteria of claim 1, the microbial inoculum of any one of claims 2-4, and/or the salt-tolerant heterotrophic nitrification-aerobic denitrification microbiome of any one or more of claims 5-6, for:
a) Waste water treatment;
b) Preparing an organic fertilizer;
c) Soil improvement;
d) Treating water eutrophication;
e) Repairing water pollution;
f) Biological denitrification.
11. The use according to claim 10, wherein,
In said c), said soil comprises saline-alkali soil; the soil improvement is soil fertility improvement;
in the d), the body of water includes fresh water and seawater.
12. The use of claim 11, wherein the soil comprises a saline-alkali soil contaminated with heavy metals including lead and cadmium.
13. A method for integrated, resource conversion of nutrients in a high salt environment, the method comprising: applying the heterotrophic nitrification-aerobic denitrification bacterium of claim 1, the microbial inoculum of any one of claims 2-4, and/or the salt-tolerant heterotrophic nitrification-aerobic denitrification microbiome of any one of claims 5-6 to a high-salt environment;
the high salt environment comprises a high salt water environment containing heavy metal pollution; the salinity of the high-salinity water environment is not lower than 3%, and the heavy metals comprise lead and cadmium.
14. The method of claim 13, wherein the high brine environment has a salinity of 3% to 8%.
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| CN110655197A (en) * | 2018-06-29 | 2020-01-07 | 龙岩学院 | Method for treating nitrate nitrogen wastewater by using heterotrophic nitrification-aerobic denitrification pseudomonas strain |
| CN113403234A (en) * | 2021-02-01 | 2021-09-17 | 山东大学 | Marine self-flocculating bacterium and halophilic nitrogen assimilation microorganism group driven to develop by marine self-flocculating bacterium as well as construction method and application of marine self-flocculating bacterium and halophilic nitrogen assimilation microorganism group |
| CN113684154A (en) * | 2021-09-08 | 2021-11-23 | 青岛蔚蓝赛德生物科技有限公司 | Pseudomonas Kunmingensis strain and application thereof in environment-friendly water treatment |
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| CN110655197A (en) * | 2018-06-29 | 2020-01-07 | 龙岩学院 | Method for treating nitrate nitrogen wastewater by using heterotrophic nitrification-aerobic denitrification pseudomonas strain |
| CN110217895A (en) * | 2019-05-31 | 2019-09-10 | 中国科学院微生物研究所 | A kind of complex micro organism fungicide and its application for water environment treatment |
| CN113403234A (en) * | 2021-02-01 | 2021-09-17 | 山东大学 | Marine self-flocculating bacterium and halophilic nitrogen assimilation microorganism group driven to develop by marine self-flocculating bacterium as well as construction method and application of marine self-flocculating bacterium and halophilic nitrogen assimilation microorganism group |
| CN113684154A (en) * | 2021-09-08 | 2021-11-23 | 青岛蔚蓝赛德生物科技有限公司 | Pseudomonas Kunmingensis strain and application thereof in environment-friendly water treatment |
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