CN106434424A - Vibrio with contaminated seawater denitrification ability and application thereof - Google Patents

Vibrio with contaminated seawater denitrification ability and application thereof Download PDF

Info

Publication number
CN106434424A
CN106434424A CN201610723504.5A CN201610723504A CN106434424A CN 106434424 A CN106434424 A CN 106434424A CN 201610723504 A CN201610723504 A CN 201610723504A CN 106434424 A CN106434424 A CN 106434424A
Authority
CN
China
Prior art keywords
vibrios
seawater
dirty
denitrification ability
strain
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201610723504.5A
Other languages
Chinese (zh)
Other versions
CN106434424B (en
Inventor
任华峰
陈进斌
苗英霞
邱金泉
成玉
王静
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Tianjin Institute of Seawater Desalination and Multipurpose Utilization SOA
Original Assignee
Tianjin Institute of Seawater Desalination and Multipurpose Utilization SOA
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Tianjin Institute of Seawater Desalination and Multipurpose Utilization SOA filed Critical Tianjin Institute of Seawater Desalination and Multipurpose Utilization SOA
Priority to CN201610723504.5A priority Critical patent/CN106434424B/en
Publication of CN106434424A publication Critical patent/CN106434424A/en
Application granted granted Critical
Publication of CN106434424B publication Critical patent/CN106434424B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • 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
    • 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
    • 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
    • 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
    • C02F2101/00—Nature of the contaminant
    • C02F2101/10—Inorganic compounds
    • C02F2101/16—Nitrogen compounds, e.g. ammonia
    • C02F2101/163—Nitrates
    • 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
    • C02F2101/166—Nitrites
    • 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
    • C02F2103/08—Seawater, e.g. for desalination

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Microbiology (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • Water Supply & Treatment (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Environmental & Geological Engineering (AREA)
  • Hydrology & Water Resources (AREA)
  • Biodiversity & Conservation Biology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)

Abstract

The invention discloses vibrio with contaminated seawater denitrification ability and application thereof and relates to the technical field of treatment of contaminated seawater. The vibrio is preserved in the Common Microorganism Center of China Committee for Culture Collection of Microorganisms with the strain preservation number of CGMCC No.12311. According to the application of the vibrio for contaminated seawater denitrification, a vibrio strain is prepared into a contamination-removal microbial agent which is used for treatment of mariculture wastewater. The industrial application prospect of the vibrio is good.

Description

There is vibrios of dirty seawater denitrification ability and application thereof
Technical field
The present invention relates to the processing technology field of dirty seawater, more particularly to a kind of arc with dirty seawater denitrification ability Bacterium, and the purposes of this vibrios, and the method preparing biological denitrogenation microbial agent with this vibrios.
Background technology
At present, in highdensity cultivation system, in feed, about 75%～80% nitrogen can enter in breeding water body, and with Different degrees of accumulation in ammonia nitrogen, nitrate nitrogen, the form of nitrite nitrogen, and aquiculture animal is worked the mischief, thus Limit in the specific yield of sea-farming, and the waste water of its discharge and also contain abundant nitrogen-containing compound, seawater can be accelerated Eutrophication, causes Disaster And Prevention Measures of Red Tides, brings harm to coastal ocean ecological environment.
Dirty seawater denitrifier is briefly that a class can carry out denitrifying bacterium under aerobic condition.Its presence makes Nitrifying process and denitrification process be able in same reactor synchronous carry out, it is anti-that the product of nitration reaction can become denitrification The substrate answered, accelerates reaction speed, reduces operating cost, the OH of anti-nitration reaction release simultaneously-Nitration reaction can be compensated The alkali consuming, it is to avoid the accumulation of acidic materials, maintains stablizing of system pH value.
Known technology, such as published 201210152921.0 1 plants of cold-resistant pseudomonad Den-05 of Chinese patent application and Its screening technique and application, filter out low-temperature aerobic denitrifying bacterium using microorganism pure culture technigne from natural habitat PseudomonaspsychrophiliaDen-05, it is achieved that the preparation of biological denitrogenation microbial agent, provides for biological fortification system Efficient denitrification bacterial classification, and be shortened by the anaerobism cycle and then shorten the control methods of aeration period, improve the municipal wastewater treatment plant winter Season denitrification effect it is achieved that under aerobic condition nitrate nitrogen removal.
Further, 201410397855.2 1 plants of efficient denitrification acinetobacter calcoaceticus novel species of Chinese patent and its application, acinetobacter calcoaceticus Acinetobacterguangzhouensis YZS-X1-1 has preferable denitrification ability, can effectively remove ammonia nitrogen and nitrous Hydrochlorate nitrogen, thus environmental protection.
Further, a kind of desulfurization simultaneously of Chinese patent 200910213798.7, denitrogenation, the method for biologically treating wastewater of decolouring, Disclose a kind of desulfurization simultaneously, denitrogenation, the method for biologically treating wastewater of decolouring.
Further, the salt cholerae strain of Chinese patent 201210171297.9 1 plant height salt biological denitrificaion and its in wastewater treatment In application.
Further, the polyculture enrichment screening method of Chinese patent application 201110288612.1 ocean heterotrophic denitrifying bacteria.
The industry urgently more diversified vibrios with dirty seawater denitrification ability, to solve in dirty seawater living intensive treatment The problem existing.
Content of the invention
Problem to be solved by this invention is, provides a kind of vibrios with dirty seawater denitrification ability and application thereof, and Apply the only dirty microbial inoculum of this vibrios preparation, and the preparation method of this microbial inoculum.
One of this case purpose is to provide a kind of vibrios with dirty seawater denitrification ability, and this case is also known as dirty seawater denitrogenation arc Bacterium.
The vibrios that the present invention has dirty seawater denitrification ability takes technical scheme below to realize：
One plant of vibrios with dirty seawater denitrification ability, is preserved in China Committee for Culture Collection of Microorganisms commonly micro- Bio-Centers, it is CGMCC No.12311 that culture presevation is numbered.
The vibrios that this case has dirty seawater denitrification ability can also be realized further using following technical measures：
The aforesaid vibrios with dirty seawater denitrification ability, wherein, described bacterial strain 16S rDNA sequence is：
This case further object is to provide a kind of aforementioned vibrios purposes.
The purposes of the aforesaid vibrios with dirty seawater denitrification ability, wherein, microorganism formulation that this bacterial strain is made, use Wastewater treatment after seawater utilization, the waste water after described seawater utilization is the marine culture wastewater containing nitrogen source pollutant.
The purposes of the aforesaid vibrios with dirty seawater denitrification ability, wherein, this bacterial strain is made and uses after microorganism formulation Wastewater treatment after seawater utilization, the waste water after described seawater utilization, nitrogen source pollutant therein, with nitrate nitrogen (NO3 -- N) count, initial concentration is 150 mg/litre.
The purposes of the aforesaid vibrios with dirty seawater denitrification ability, wherein, this bacterial strain is made and uses after microorganism formulation Wastewater treatment after seawater utilization, the suitable carbon source wherein with the cholerae strain of dirty seawater denitrification ability is glucose or second Sour sodium or sodium succinate or sodium oxalate or sodium citrate any one.
The purposes of the aforesaid vibrios with dirty seawater denitrification ability, wherein, after the microorganism formulation that this bacterial strain is made For the wastewater treatment after seawater utilization, wherein, the suitable pH value of cholerae strain with dirty seawater denitrification ability is in 5.0-6.0 Or the arbitrary value range of 6.1-7.0 or 7.1-8.0 or 8.1-9.0.
The purposes of the aforesaid vibrios with dirty seawater denitrification ability, wherein, this bacterial strain is made and uses after microorganism formulation Wastewater treatment after seawater utilization, wherein, the suitable pH value of cholerae strain with dirty seawater denitrification ability is 5.0 or 6.0 Or 7.0 or 8.0 or 9.0 any value.
Thus, dirty microbial inoculum is used for marine culture wastewater only to have the microorganism that the cholerae strain of dirty seawater denitrification ability makes Process, process time only 24 hours, sewage water denitrification clean-up effect is with regard to highly significant, and microbial inoculum has reliable bio-safety Property.
This case further object is to provide a kind of microbial inoculum of the vibrios with dirty seawater denitrification ability.
A kind of vibrios microbial inoculum with dirty seawater denitrification ability, wherein, the described vibrios with dirty seawater denitrification ability protects It is hidden in China Committee for Culture Collection of Microorganisms's common micro-organisms center, it is CGMCC No.12311 that culture presevation is numbered.
The aforementioned vibrios microbial inoculum with dirty seawater denitrification ability, wherein, the described vibrios with dirty seawater denitrification ability, fit Close carbon source be glucose or sodium acetate or sodium succinate or sodium oxalate or sodium citrate any one；
The aforesaid vibrios microbial inoculum with dirty seawater denitrification ability, wherein, the described vibrios with dirty seawater denitrification ability The G+C content of DNA is 44.3mol%；Described " G+C content " refers to " shared by guanine (G) and cytimidine (C) in DNA molecular Mole percent ratio ".
The aforesaid vibrios microbial inoculum with dirty seawater denitrification ability, wherein, the temperature of suitable growth is 20 DEG C -25 DEG C or 25 DEG C -30 DEG C or 31 DEG C -35 DEG C or 36 DEG C -37 DEG C or 38-40 DEG C of arbitrary value range.
The aforementioned vibrios microbial inoculum with dirty seawater denitrification ability, wherein, the described vibrios with dirty seawater denitrification ability, fit Close pH value in the arbitrary value range of 5.0-6.0 or 6.1-7.0 or 7.1-8.0 or 8.1-9.0.
This case further object is to provide a kind of bacterial preparation process of the vibrios with dirty seawater denitrification ability.
A kind of preparation method of the vibrios microbial inoculum with dirty seawater denitrification ability, the method has inoculation in culture In base, the step of culture bacterial classification, wherein, inoculation is that this vibrios test tube kind is inoculated in beef-protein medium, Cultivation temperature is 20 DEG C -40 DEG C, hunting speed is 120-180 rev/min, and shake flask is cultivated to logarithmic phase；
The described vibrios with dirty seawater denitrification ability, is preserved in China Committee for Culture Collection of Microorganisms common Microorganism center, it is CGMCC No.12311 that culture presevation is numbered.
The preparation method of the aforesaid vibrios microbial inoculum with dirty seawater denitrification ability, also, cultured bacterial classification is accessed Step to fermentation tank culture medium：By cultured bacterial classification, by the inoculum concentration of 0.5%-1% or 1%-2% of culture medium weight It is linked in the fermentation tank of preset volume, have the fermentation tank culture medium of predetermined amount in fermentation tank, cultivate to exponential phase.
The preparation method of the aforesaid vibrios microbial inoculum with dirty seawater denitrification ability, also, the step of fermented and cultured：Bacterial classification After being linked into fermentation tank, the throughput of filtrated air is 10-30 liter/min；
The preparation method of the aforesaid vibrios microbial inoculum with dirty seawater denitrification ability, mixing speed is 120 rev/min -200 Rev/min, or/and, cultivation temperature is 25 DEG C -40 DEG C；
The preparation method of the aforesaid vibrios microbial inoculum with dirty seawater denitrification ability, whole process incubation time is that 24-48 is little When；
The preparation method of the aforesaid vibrios microbial inoculum with dirty seawater denitrification ability, nutrient solution packing after the completion of fermentation, or Using the absorption of aseptic attachment, it is dried, make solid fungicide.
The preparation method of the aforesaid vibrios microbial inoculum with dirty seawater denitrification ability, wherein, by inoculation in culture medium In, the step of culture bacterial classification, wherein, described beef-protein medium has：Peptone 1%, beef extract 0.3%, chlorine Change sodium 0.5%, pH 7.0, balance of additive.
The preparation method of the aforesaid vibrios microbial inoculum with dirty seawater denitrification ability, wherein, cultured bacterial classification is accessed Step to fermentation tank culture medium：Described fermentation tank culture based component has：Na2HPO4、KH2PO4、KNO3, glucose and seawater； When seawater amount is 1000mL, Na2HPO4For 7.9g, KH2PO4For 1.5g, KNO3For 1.0g, glucose is 9.4g, and fermentation tank is trained Foster base pH value is 7.0-7.5.
The preparation method of the aforesaid vibrios microbial inoculum with dirty seawater denitrification ability, wherein, by inoculation in culture medium In, culture bacterial classification step, wherein, cultivation temperature be 20 DEG C -25 DEG C or 25 DEG C -30 DEG C or 31 DEG C -35 DEG C or 36 DEG C -37 DEG C or 38-40 DEG C of arbitrary value range.
The preparation method of the aforesaid vibrios microbial inoculum with dirty seawater denitrification ability, wherein, by inoculation in culture medium In, the step of culture bacterial classification, wherein, hunting speed is 120-150 rev/min or 151-160 rev/min or 160-170 rev/min Arbitrary value range of clock or 171-180 rev/min.
The preparation method of the aforesaid vibrios microbial inoculum with dirty seawater denitrification ability, wherein, the step of fermented and cultured：Bacterial classification It is linked into fermentation tank after fermentation culture, filtrated air throughput is preferably 10 liters/min or 20 liters/min or 30 liters/min Any value；
The preparation method of the aforesaid vibrios microbial inoculum with dirty seawater denitrification ability, mixing speed is preferably 60-90 rev/min Arbitrary value range of clock or 91-120 rev/min；
The preparation method of the aforesaid vibrios microbial inoculum with dirty seawater denitrification ability, cultivation temperature is preferably 25 DEG C -30 DEG C Or 31 DEG C -35 DEG C or 36 DEG C -40 DEG C of arbitrary value range；
The preparation method of the aforesaid vibrios microbial inoculum with dirty seawater denitrification ability, described whole process incubation time is preferably 24-30 hour or 31-36 hour or 37-42 hour or the arbitrary value range of 43-48 hour；
The preparation method of the aforesaid vibrios microbial inoculum with dirty seawater denitrification ability, described dirt seawater denitrogenation dirty bacterium only, fit Close pH value in the arbitrary value range of 5.0-6.0 or 6.1-7.0 or 7.1-8.0 or 8.1-9.0；
The preparation method of the aforesaid vibrios microbial inoculum with dirty seawater denitrification ability, described dirt seawater denitrogenation bacteria strain suitable Close carbon source be glucose or sodium acetate or sodium succinate or sodium oxalate or sodium citrate any one.
The present invention compared with prior art has significant advantage and beneficial effect.From above technical scheme, this Bright under excellent structure configures, at least following advantage：
This case has vibrios of dirty seawater denitrification ability and application thereof, the vibrios MCW153 in the present invention, not only has relatively Good denitrification ability, can effectively remove nitrate nitrogen and nitrite nitrogen, and safe and harmless, and biological safety test shows (thalline feeds goldfish 3 months), goldfish upgrowth situation is good, and is better than control group.Vibrios MCW153 is used for making biological denitrificaion Microbial inoculum, for solving, seawater aquaculture system total nitrogen is exceeded to wait eutrophication problem, has good industry application and market Prospect.
This case dirt seawater denitrifier be successfully separated can solve marine culture wastewater process long felt a difficult problem, this Case dirt seawater denitrifier separation method is simple, separates and cultural method is relatively easy to promote, with the development of culture technique, it can So that by the use of many organic matters (including fishes and shrimps excreta, food-residue etc.) as its carbon source, therefore, dirt seawater denitrifier makees for this case For the biomaterial of the efficient dirty microbial bacterial agent only of dirty seawater, there are excellent industry promotion and application and be worth.
Be there is by this case the microbial inoculum of the vibrios preparation of dirty seawater denitrification ability, the manufacturing cost more cheap of microbial inoculum can be made, more It is beneficial to industry popularization and application；The microbial inoculum being obtained by the bacterial preparation process of this case especially can meet the purification of marine culture wastewater The practical problem processing and demand, can quickly start the biological cleaning ability of newly-built saline sewage treatment facility using this microbial inoculum With the strengthened denitrification disposal ability recovering the biosystem deteriorating.
The vibrios MCW153 that one plant of this case has dirty seawater denitrification ability has good total nitrogen removal effect, and laboratory is shaken Bottle test result shows, manual simulation's waste water nitrogen removal rate is higher.
It can be seen that, vibrios described in this case is prepared into the microorganism formulation administered for marine culture wastewater, for current routine Sewage disposal technology can not process the situation of marine culture wastewater (low-carbon high-nitrogen) effectively, using the biological reinforced skill of this case Art, by adding the microbial inoculum of this case vibrios in sewage treatment facility, is substantially shorter the sludge acclimatization time, significantly improves denitrogenation Treatment effect, can effectively reduce investment outlay and operating cost, thus solving de- present in current marine culture wastewater biological treatment Nitrogen effect difference and the problem in the urgent need to address such as sludge acclimatization period is long.
Present invention contrast prior art have significant contribute and progressive, strictly there is novelty, creativeness, practicality Good technology.
Brief description
Fig. 1 is vibrios thalline transmission electron microscope photo of the present invention；
Fig. 2 is vibrios optimum temperature schematic diagram of the present invention；
Fig. 3 is the most suitable rotating speed schematic diagram of vibrios of the present invention；
Fig. 4 is the most suitable inoculum concentration schematic diagram of vibrios of the present invention；
Fig. 5 is the most suitable throughput schematic diagram of vibrios of the present invention；
Fig. 6 is vibrios of the present invention optimal fermentation time schematic diagram；
Fig. 7 is vibrios optimum pH schematic diagram of the present invention；
Fig. 8 is the most suitable carbon source schematic diagram of vibrios of the present invention；
Fig. 9 is the denitrification effect schematic diagram to artificial seawater breeding wastewater for the microbial inoculum of the present invention；
Figure 10 is the biological safety test of the vibrios MCW153 that one plant of this case has dirty seawater denitrification ability.
Specific embodiment
The specific embodiment of the present invention is shown in detail in by following examples.
Below in conjunction with preferred embodiment, to the specific embodiment providing according to the present invention, step, feature and its effect, Describe in detail as follows.
Embodiment 1
A kind of vibrios with dirty seawater denitrification ability, this case is also referred to as dirty seawater denitrifier, and it is micro- that this vibrios is deposited in China Biological inoculum preservation administration committee common micro-organisms center, it is CGMCC No.12311 that culture presevation is numbered.
The G+C content of the described vibrios DNA with dirty seawater denitrification ability is 44.3mol%.There is dirty seawater denitrogenation The vibrios of ability, Gram-negative (G-), cell is oval, with polar flagella motion.There is dirty seawater denitrification ability Vibrios, oxidizing ferment, catalase, any one reduction reaction of nitrate are the positive.There is the vibrios bacterium of dirty seawater denitrification ability Strain is carbon source using glucose, sodium acetate, sodium succinate etc..Have dirty seawater denitrification ability cholerae strain utilize glucose and Other (galactolipin, arabinose, maltose etc.) some carbohydrates produce acid.The suitable pH value of dirty seawater denitrifier, between 5-9, is fitted Preferably growth temperature is 30-35 DEG C.
The vibrios MCW153 that one plant of this case has dirty seawater denitrification ability has good total nitrogen removal effect, and laboratory is shaken Bottle test result shows, manual simulation's waste water nitrogen removal rate is higher.
Embodiment 2
There is the only dirty bacterial preparation process of the vibrios of dirty seawater denitrification ability, including following technique mistake described in embodiment 1 Journey：
Step 1：The vibrios test tube kind with dirty seawater denitrification ability is inoculated in beef-protein medium, culture Temperature is 30 DEG C, hunting speed is 160 revs/min, and shake flask is cultivated to logarithmic phase；Beef-protein medium is commercially available Culture medium.
Step 2：By above-mentioned cultured bacterial classification, it is linked in fermentation tank by the inoculum concentration of culture medium weight 0.5%-2%, There is culture medium in fermentation tank；Culture medium used by fermentation tank has：Na2HPO47.9 grams, KH2PO41.5 grams, KNO31.0 grams, grape 9.4 grams of sugar, 1000 milliliters of seawater.
Step 3：In fermentation tank production process, the throughput of filtrated air is 10-30 liter/min, and mixing speed is 160 Rev/min, cultivation temperature is 30 DEG C, and whole process incubation time is 24 hours, nutrient solution packing after the completion of fermentation, or using aseptic Attachment adsorbs, is dried, and makes solid fungicide.
As can be seen here, the vibrios separation method with dirty seawater denitrification ability is simple, separates and culture is relatively easy to, with The development of culture technique, it can be by the use of many organic matters (including fishes and shrimps excreta, food-residue etc.) as its carbon source.
Microbial inoculum, manufacturing cost more cheap are prepared by the vibrios with dirty seawater denitrification ability, the industry that is relatively beneficial to is promoted should With；
Thus, there is the bacteria agent that the vibrios bacterial preparation process of dirty seawater denitrification ability obtains, especially can meet sea The practical problem of the purified treatment of water breeding wastewater and demand, can quickly start newly-built saline sewage process using this microbial inoculum and set The strengthened denitrification disposal ability of the biosystem of the biological cleaning ability applied and recovery deterioration.
Embodiment 3
A kind of purposes of the vibrios with dirty seawater denitrification ability, is that dirty microbial inoculum is used for sea to the microorganism made of vibrios strain only Water cultivating wastewater purification, process time is 24 hours, and sewage water denitrification clean-up effect is notable.
Embodiment 4
Referring to accompanying drawing 1 to shown in Figure 10, this case is related to component and quotes percentage by weight, under do not repeat.
One plant of vibrios MCW153 with dirty seawater denitrification ability, this biomaterial is preserved in Chinese microorganism strain preservation Administration committee's common micro-organisms center, it is CGMCC No.12311 that culture presevation is numbered, March 25 2016 preservation time, Survival, is shown in that preservation proves；Fig. 1 is bacterial strain MCW153 transmission electron microscope photo；
In this case, the vibrios MCW153 with dirty seawater denitrification ability, also known as dirty seawater denitrifier, is described in detail below：
This dirty seawater denitrifier is identified as vibrio (Vibrio sp.MCW153), and Main Biological is gram Negative (G-), cell is ellipse, amphitrichous, 0.8 × 1.2 micron of thalline size；The culture bacterium of 3 days on LB culture medium flat plate Fall moisten for cream-coloured, flattened round, surface is slight graininess, edge slight fold, diameter is in 2.5 millimeter.
The described vibrios with dirty seawater denitrification ability, oxidizing ferment, catalase, any one reduction reaction of nitrate are Positive；Its urease test, indole reaction are feminine gender；Using some carbohydrates such as glucose, galactolipin, arabinose, maltose Produce acid.Referring to dirty seawater denitrogenation dirty bacterium MCW153 physiological and biochemical property table only.The described vibrios with dirty seawater denitrification ability is fitted Between 5-9, preferably suitable growth temperature is 30 DEG C to suitable pH value.The G+C of the described vibrios DNA with dirty seawater denitrification ability Content is 44.3mol%；Wherein, " G+C content " is to be often referred to " shared by guanine (G) and cytimidine (C) in DNA molecular Mole percent ratio ".
Dirty seawater denitrifier MCW153 physiological and biochemical property
The Genbank number of logging in of this bacterial strain 16S rDNA is KU644134.
Carry out PCR (PCR) amplification, polymerase chain reaction to separating the bacterial strain 16S rDNA sequence obtaining Sea raw work sequencing should be served by (PCR) product, base sequence is as follows：
The bacterial strain 16S rDNA sequence of dirty seawater denitrogenation vibrios is：
NCBI the sequencing results show, the 16S rDNA sequence of MCW153 bacterial strain and the multiple bacterium of Vibrio sp. (vibrios) The sequence homology of strain is more than 99%.
There is the preparation method of the vibrios microbial inoculum of dirty seawater denitrification ability, including following technical process：
Step 1：The vibrios test tube kind with dirty seawater denitrification ability is inoculated in beef-protein medium, culture Temperature is 25 DEG C -40 DEG C, hunting speed is 120-180 rev/min, and shake flask is cultivated to logarithmic phase；Beef extract-peptone is trained Foster base can be general commercially available product；
Step 1 further embodiment is：
Preferably beef-protein medium has：Peptone 1%, beef extract 0.3%, sodium chloride 0.5%, pH 7.0, it will not go into details for balance of usual additive；
The described vibrios with dirty seawater denitrification ability is preserved in Chinese microorganism strain preservation conservator for aforesaid The bacterial classification of meeting common micro-organisms center, referring to Fig. 1, deposit number is CGMCC No.12311；
The described vibrios test tube kind with dirty seawater denitrification ability is inoculated in the culture temperature in beef-protein medium Degree is preferably 20 DEG C -25 DEG C or 26 DEG C -30 DEG C or 31 DEG C -35 DEG C or 36 DEG C -37 DEG C or 38-40 DEG C of arbitrary value range；
Referring to Fig. 2, test shows, temperature be 20 DEG C～40 DEG C when, bacterial strain can well-grown, and temperature be 30 DEG C When, biomass reaches maximum.
The described vibrios test tube kind with dirty seawater denitrification ability is inoculated in the shaking flask in beef-protein medium The speed of vibration is preferably 120-150 rev/min or 151-160 rev/min or 161-170 rev/min or 171-180 rev/min Arbitrary value range of clock, mixing speed rev/min also is indicated as r/min；
Referring to Fig. 3, test shows the impact to strain growth for the shaking speed, and the speed of shake flask is too low to be unfavorable for bacterial strain The growth of MCW153, with faster rotational speed, cellular biomass increases, but too high rotating speed easily makes nutrient solution overflow, and therefore, fits Suitable strain culturing rotating speed is 120-180 rev/min, and optimal rotating speed is 160 revs/min.
Step 2：By above-mentioned cultured bacterial classification, it is linked into default appearance by the inoculum concentration of the 0.5%-2% of culture medium weight In long-pending fermentation tank, there is the culture medium of predetermined amount in fermentation tank, such as by above-mentioned cultured bacterial classification, the inoculum concentration by 1% accesses To in the fermentation tank of preset volume amount, such as 10 liters of fermentation tanks, cultivate to exponential phase；
Step 2 further embodiment is：
Preferably the culture medium used by fermentation tank has：Na2HPO47.9 grams, KH2PO41.5 grams, KNO31.0 grams, glucose 9.4 grams, pH 7.0-7.5,1000 milliliters of seawater；
The described inoculum concentration of vibrios with dirty seawater denitrification ability is preferably 1%- with the percentage by weight of culture medium 1.5% or 1.5%-2% arbitrary value range；
Referring to Fig. 4, this test indicates the inoculum concentration of the vibrios with dirty seawater denitrification ability to the impact preparing microbial inoculum. After cultivating 24 hours, increase with inoculum concentration, less, therefore, the suitable inoculum concentration of bacteria fermentation exists the change of microbial inoculum increment 1-2%, but combine cost factor, selects 1% inoculum concentration can effective reduces cost；
Step 3：In fermentation tank production process, the throughput of filtrated air is 10-30 liter/min, and mixing speed is 120 Revs/min -200 revs/min, cultivation temperature is 25 DEG C -40 DEG C, and whole process incubation time is 24-48 hour, trains after the completion of fermentation Nutrient solution dispenses, or is adsorbed using aseptic attachment, is dried, and makes solid fungicide.
Throughput liter/min also is indicated as L/min.
Step 3 further embodiment is：
Described filtrated air throughput is preferably 10 liters/min or 20 liters/min or 30 liters/min of arbitrary value range；
Referring to Fig. 5, test shows that the impact to strain fermentation for the throughput is larger, and wherein, throughput is 10-30 liter/min When, microbial inoculum well-grown.
Mixing speed is preferably 60-90 rev/min or 91-120 rev/min of arbitrary value range；
Cultivation temperature is preferably 25 DEG C -30 DEG C or 31 DEG C -35 DEG C or 36 DEG C -40 DEG C of arbitrary value range；
Described whole process incubation time is preferably 24-30 hour or 31-36 hour or 37-42 hour or 43-48 is little former One value range；Hour also is indicated as h.
Referring to Fig. 6, test shows, with the increase of fermented incubation time, thalline constantly grows, and wherein, fermentation time is 4-16 hour, microbial inoculum is in optimal growth conditions.
The culture medium of described fermentation tank includes：Na2HPO47.9 grams of (disodium hydrogen phosphate), KH2PO4(potassium dihydrogen phosphate) 1.5 grams, KNO31.0 grams of (potassium nitrate), 9.4 grams of glucose, pH 7.0-7.5,1000 milliliters of seawater；
The described vibrios with dirty seawater denitrification ability, suitable pH value is preferably in 5.0-6.0 or 6.1-7.0 or 7.1-8.0 Or the arbitrary value range of 8.1-9.0.
The described vibrios with dirty seawater denitrification ability, optimum pH is preferably in any value of 6 or 7 or 8 or 9；
Referring to Fig. 7, it is wider, in the equal energy of initial pH value 5.0-9.0 scope that test shows that bacterial strain MCW153 adapts to pH value range Interior energy well grows.
Referring to Fig. 8, test shows carbon source (glucose, sodium acetate, sodium succinate, sodium oxalate, sodium citrate etc.) to microbial inoculum Fermentation has large effect, and wherein, glucose is that nitrate nitrogen removal effect during carbon source is optimal, and therefore, glucose is bacterial strain The most suitable carbon source of MCW153.
Acceptable after the completion of fermentation：Nutrient solution is directly dispensed with plastic barrel, or makes solid using charcoal absorption Microbial inoculum.
Microbial inoculum is obtained by preceding method there is reliable biological safety.
Referring to Figure 10, biological safety test (thalline feeds goldfish) shows, vibrios MCW153 bacterial strain, safe and harmless, bacterium Body feeds goldfish 3 months, and goldfish upgrowth situation is good, and is better than control group.
The process that microbial inoculum is used for marine culture wastewater is obtained by preceding method, sewage water denitrification clean-up effect is notable.
Marine culture wastewater is applied to by the vibrios microbial inoculum with dirty seawater denitrification ability that foregoing embodiments are obtained Process, and constantly Measures program prepared by further microbial inoculum, can make the manufacturing cost more cheap of microbial inoculum, the industry of being relatively beneficial to pushes away Wide apply, have following tests can illustrate that dirty microbial inoculum processes the excellent effect of marine culture wastewater only for this case dirt seawater denitrogenation.
Laboratory shake flask is tested, referring to Fig. 9：
MCW153 inoculation in nitrate nitrogen (NO3 -- N) initial concentration be 150 mg/litre artificial seawater cultivation useless In water, 30 DEG C, 160 revs/min of shaking table cultures, timing sampling measures thalli growth, nitrate nitrogen (NO3 -- N) and nitrite nitrogen (NO2 -- N) concentration levels.
The result of Fig. 9 shows, MCW153 can be by nitrate nitrogen (NO in 12 hours3 -- N) concentration is more than 150 mg/litre It is down to below 0.05 mg/litre, meanwhile, nitrite nitrogen (NO2 -- N) concentration be first stepped up, then gradually reduce to 0.05 Below mg/litre, illustrate that MCW153 bacterial strain has efficient denitrification denitrogenation ability, artificial seawater breeding wastewater is had preferably Denitrogenation clean-up effect.
In addition, by Fig. 9 it is also shown that the growth of MCW153 bacterial strain is synchronized with nitrate nitrogen (NO3 -- N) concentration reduction, and The growth curve of MCW153 bacterial strain, nitrite nitrogen (NO2 -- N) metabolic chart in lag phase substantially all in 0-3 hour, so Enter logarithmic phase (4-15 hour) afterwards, about reach highest thalli growth amount at 24 hours.Concentration unit " mg/litre " also illustrates that For " mg/L ".Waste water after seawater utilization is the marine culture wastewater containing nitrogen source pollutant.
This case has vibrios of dirty seawater denitrification ability and application thereof, the vibrios MCW153 in the present invention, not only has relatively Good denitrification ability, can effectively remove nitrate nitrogen and nitrite nitrogen, and safe and harmless, and biological safety test shows (thalline feeds goldfish 3 months), goldfish upgrowth situation is good, and is better than control group.Vibrios MCW153 can be used for making further Make biological denitrogenation microbial agent, seawater aquaculture system total nitrogen is exceeded to wait eutrophication problem particularly with solving, and has good Application prospect.
The above, be only presently preferred embodiments of the present invention, and not the present invention is made with any pro forma restriction, all It is any simple modification, equivalent variations and modification above example made according to the technical spirit of the present invention, all still fall within In the range of technical solution of the present invention.

Claims (7)

1. one plant of vibrios with dirty seawater denitrification ability, is preserved in China Committee for Culture Collection of Microorganisms's commonly micro- life Thing center, it is CGMCC No.12311 that culture presevation is numbered.
2. there is the vibrios of dirty seawater denitrification ability as claimed in claim 1, wherein, described bacterial strain 16SrDNA sequence is：
3. the purposes of the vibrios with dirty seawater denitrification ability according to claim 1 and 2 is it is characterised in that by this bacterium The microorganism formulation that strain is made, for the wastewater treatment after seawater utilization, the waste water after described seawater utilization is dirty containing nitrogen source The marine culture wastewater of dye thing.
4. the purposes of the vibrios with dirty seawater denitrification ability according to claim 1 and 2 is it is characterised in that by this bacterium For the wastewater treatment after seawater utilization, the waste water after described seawater utilization, nitrogen source therein after the microorganism formulation that strain is made Pollutant, with nitrate nitrogen (NO3 -- N) meter, initial concentration is 150 mg/litre.
5. the purposes of the vibrios with dirty seawater denitrification ability according to claim 3 or 4 is it is characterised in that by this bacterium For the wastewater treatment after seawater utilization after the microorganism formulation that strain is made, wherein there is the cholerae strain of dirty seawater denitrification ability Suitable carbon source be glucose or sodium acetate or sodium succinate or sodium oxalate or sodium citrate any one.
6. the purposes of the vibrios with dirty seawater denitrification ability according to claim 3 or 4 is it is characterised in that by this bacterium For the wastewater treatment after seawater utilization after the microorganism formulation that strain is made, wherein, there is the vibrios bacterium of dirty seawater denitrification ability The suitable pH value of strain is in the arbitrary value range of 5.0-6.0 or 6.1-7.0 or 7.1-8.0 or 8.1-9.0.
7. the purposes of the vibrios with dirty seawater denitrification ability according to claim 3 or 4 is it is characterised in that by this bacterium For the wastewater treatment after seawater utilization after the microorganism formulation that strain is made, wherein, there is the vibrios bacterium of dirty seawater denitrification ability The suitable pH value of strain is in 5.0 or 6.0 or 7.0 or 8.0 or 9.0 any value.
CN201610723504.5A 2016-08-23 2016-08-23 Vibrios and application thereof with dirty seawater denitrification ability Expired - Fee Related CN106434424B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610723504.5A CN106434424B (en) 2016-08-23 2016-08-23 Vibrios and application thereof with dirty seawater denitrification ability

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610723504.5A CN106434424B (en) 2016-08-23 2016-08-23 Vibrios and application thereof with dirty seawater denitrification ability

Publications (2)

Publication Number Publication Date
CN106434424A true CN106434424A (en) 2017-02-22
CN106434424B CN106434424B (en) 2019-09-06

Family

ID=58182970

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610723504.5A Expired - Fee Related CN106434424B (en) 2016-08-23 2016-08-23 Vibrios and application thereof with dirty seawater denitrification ability

Country Status (1)

Country Link
CN (1) CN106434424B (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108342341A (en) * 2018-03-06 2018-07-31 武汉光谷环保科技股份有限公司 Cholerae strain for handling high nitrogen low-carbon brine waste and its application
CN108441439A (en) * 2018-03-06 2018-08-24 武汉光谷环保科技股份有限公司 Cholerae strain and cultural method for handling high nitrogen low-carbon brine waste

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102747014A (en) * 2012-05-29 2012-10-24 北京盖雅技术中心有限公司 High-saline biological denitrification salinivibrio strain and application thereof in wastewater treatment

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102747014A (en) * 2012-05-29 2012-10-24 北京盖雅技术中心有限公司 High-saline biological denitrification salinivibrio strain and application thereof in wastewater treatment

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
苏兵: "一株海洋异养硝化—好氧反硝化菌的分离鉴定、脱氮特性及应用", 《中国优秀硕士学位论文全文数据库 基础科学辑》 *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108342341A (en) * 2018-03-06 2018-07-31 武汉光谷环保科技股份有限公司 Cholerae strain for handling high nitrogen low-carbon brine waste and its application
CN108441439A (en) * 2018-03-06 2018-08-24 武汉光谷环保科技股份有限公司 Cholerae strain and cultural method for handling high nitrogen low-carbon brine waste
CN108441439B (en) * 2018-03-06 2022-03-29 武汉光谷环保科技股份有限公司 Vibrio strain for treating high-nitrogen low-carbon salt-containing wastewater and culture method

Also Published As

Publication number Publication date
CN106434424B (en) 2019-09-06

Similar Documents

Publication Publication Date Title
CN101565240B (en) Method for preparing water quality modifying microecological preparation for reducing ammonia nitrogen and nitrite nitrogen
CN101565239A (en) Method for preparing composite heterotrophic nitrobacteria water quality modifying solid powder or granule
CN104845920A (en) Marine zobellella sp. and application thereof
WO2021077453A1 (en) Pseudomonas stutzeri, composite microbial inoculum prepared using the pseudomonas stutzeri, and use of the composite microbial inoculum
CN115125164B (en) Highly salt-resistant and heavy metal-resistant heterotrophic nitrification-aerobic denitrification self-flocculation marine bacterium and application thereof in community construction
CN114703095A (en) Pseudomonas mendocina and application thereof in field of sewage and wastewater purification
CN101811779B (en) Preparation method of halophilic decontamination bacterial agent and bacterial agent prepared by same
CN101701197B (en) Novel microorganism flora mixture and mixed nutrient medium thereof
CN113151063B (en) Citrobacter freundii AS11 and application thereof in sewage treatment
CN108546657B (en) Baria alismoides FX-1 and application thereof
CN111690546B (en) Rhodosporidium toruloides ZDFY1801 and application thereof
CN106434424B (en) Vibrios and application thereof with dirty seawater denitrification ability
CN109055252A (en) A kind of heterotrophic nitrification-aerobic denitrification complex microorganism preparations and preparation method thereof
CN111471611A (en) Rhodococcus ruber HDRR1 for purifying inorganic nitrogen and phosphorus in tail water of seawater pond culture and application thereof
CN106434423A (en) Preparation method of contaminated seawater denitrification vibrio microbial agent and application of microbial agent
CN106434422A (en) Preparation method of vibrio bacterial agent with contaminated seawater denitrification ability and bacterial agent thereof
CN101811780B (en) Preparation method and application of halophilic decontamination bacterial agent
CN111471612B (en) Rhodococcus ruber HDRR2Y for purifying inorganic nitrogen and phosphorus in seawater pond culture tail water and application thereof
CN110499265B (en) Autotrophic nitrifying bacteria consortium and culture expanding method thereof
CN108728372B (en) Sphingomonas bacterium LPN080 capable of assimilating ammonia monoxide, microbial preparation and application thereof
CN110184217B (en) Salt-tolerant denitrifying bacterium taking nitrite as nitrogen source and application thereof
CN108546662A (en) Using the method for nitrifying bacteria community-bacillus Combined Treatment breeding wastewater of immobilization respectively
US10400255B2 (en) Method of converting marine fish waste to biomethane
CN116396912B (en) Pyrethroid pesticide intermediate degrading bacterium, microbial inoculum and wastewater treatment method and treatment device thereof
CN116904349B (en) Adhesive sword bacteria with aerobic denitrification capability and application thereof

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
CB02 Change of applicant information

Address after: 300 000 Nankai District, Tianjin, No. 55 Navigation Channel

Applicant after: Tianjin Institute of Seawater Desalination and Comprehensive Utilization, Ministry of Natural Resources

Address before: 300192 No. 1 East scientific research road, Tianjin, Nankai District

Applicant before: Tianjin Inst. of Seawater Desalination and Comprehensive Application, National S

CB02 Change of applicant information
GR01 Patent grant
GR01 Patent grant
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20190906

Termination date: 20200823

CF01 Termination of patent right due to non-payment of annual fee