CN107699513A - A kind of special degradation bacteria of black and odorous water and its application - Google Patents
A kind of special degradation bacteria of black and odorous water and its application Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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
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- C12R2001/11—Bacillus megaterium
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- 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
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/105—Phosphorus compounds
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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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Abstract
The invention provides a kind of special degradation bacteria strains of black and odorous water and its application in black and odorous water is administered, the bacterial strain is bacillus megaterium, deposit number CGMCC No.13543, the bacterial strain reduce the COD of black and odorous water, the ammonia nitrogen in degraded black and odorous water and/or remove phosphate.The special degradation bacteria strains of black and odorous water provided by the invention, there is strong selectivity to the pollutant in black and odorous water, the organic matter and ammonia nitrogen in black and odorous water can efficiently be utilized, the COD of black and odorous water can reduce by more than 90%, good phosphorus ligands ability is provided simultaneously with, black and odorous water purification of water quality efficiency can be greatly improved.
Description
Technical field
The invention belongs to microorganism field, and in particular to a kind of special degradation bacteria of black and odorous water and its application.
Background technology
Black and odorous water is that unpleasant color (black and blacking up color) is presented and (or) gives out that to make us foreign odor (smelly
Or stench) water body general name.In recent years, China's major part urban river, closing are received with semiclosed swag water body in the presence of excessive
Dirty situation.Excess emissions directly result in body eutrophication, and water body is unbalance for oxygen consumption, ammonia nitrogen, hydrogen sulfide, iron, manganese sulfide etc.
Concentration raises, so as to cause water quality constantly to deteriorate and occur with black smelly phenomenon.Water body organism pollution is to cause black and odorous water
One of important factor in order.Organic matter such as carbohydrate, amino acid, protein, grease, esters etc. are mainly with solubilised state, suspended state
In the presence of these materials resolve into small-molecule substance under microbial action, and this process will consume a large amount of dissolved oxygens.In black and odorous water
Primarily serve blackening effect is inorganic pollution, wherein main blackening composition is easily oxidized FeS and MnS.Chemical oxygen demand
Amount (COD) reflects the degree polluted in water by reducing substances, and these materials include organic matter, nitrite, ferrous salt, sulphur
Compound etc., therefore, COD are a composite targets of reducing substances relative amount.
Black and odorous water Treatment process mainly includes artificial aeration, ecological engineering reparation, biological growth-promoting, microbial cleaning, group
The technology such as symphysis thing and plant purification.Wherein, using special microorganism come be metabolized high density pollution thing be at present both at home and abroad administer
The main method and development trend of black and odorous water.Efficient special degradation bacteria can be in serious pollution water body by using organic
Thing, hydrogen sulfide, ammonia etc. synthesize the component of itself as hydrogen donor, increase dissolved oxygen, so as to play a part of purifying sewage.
At present, special microorganism strain limited amount, the degradation effect of black and odorous water are uneven, related process technologies are to be developed.
In Related product, efficiently special bacteria agent needs import for part, improves cost, have impact on the fast development in technological service market.Cause
This, it would be highly desirable to the special microorganism fungus kind of black and odorous water and microbial inoculum are developed as early as possible with reference to China's water body treating real needs, with service
Municipal relevant departments and technical market.
The content of the invention
In order to obtain the new microbial inoculum processing system of Effec-tive Function, inventor has done substantial amounts of work, black smelly for China
Water body characteristic feature, according to microorganisms in water NATURAL DISTRIBUTION and metabolic rule, efficient water body is separated and filtered out from sludge
Contaminant degradation bacterial strain.
Therefore, the invention provides a kind of special degradation bacteria strains of black and odorous water, the bacterial strain is bacillus megaterium, name
For AW0F5, Chinese microorganism strain preservation management committee of the institute 3 of Chaoyang District, Beijing City Beichen Lu 1 is preserved on January 10th, 2017
Member's meeting common micro-organisms center, preserving number is CGMCC No.13543.AW0F5 16S rDNA sequences such as SEQ ID NO.1 institutes
Show.
The screening step of above-mentioned bacterial strains is in the present invention：Sludge, sample collection are taken from Jizhuangzi Waste Water Treatment Plant, Tianjin
Sterilization container is placed at once afterwards to take back；5g sludge is taken to shake expansion culture 24h in LB fluid nutrient mediums under aseptic condition；Take
5mL nutrient solutions are transferred cultivates 24h (being repeated 3 times) in LB fluid nutrient mediums；Last time LB nutrient solutions 1mL is taken to dilute 103-108
Times, dilution is coated on No. 1 solid medium of Gao Shi；30 DEG C incubated, line separation repeatedly occurs after bacterium colony to going out
Existing pure single bacterium colony.
Present invention also offers a kind of method for expanding the foregoing special degradation bacteria strains of black and odorous water, it is trained using LB liquid
Support base；Cultivation temperature is 5-55 DEG C, preferably 30-40 DEG C；PH is 6.8-8.0, preferably 7.2-7.8.
Present invention also offers another method for expanding the foregoing special degradation bacteria strains of black and odorous water, it uses Gao Shi 1
Number fluid nutrient medium；Cultivation temperature is 15-45 DEG C, preferably 25-40 DEG C；PH is 6.8-7.5.
The invention provides application of the special degradation bacteria strains of black and odorous water in black and odorous water is administered.
In one embodiment of the invention, the special degradation bacteria strains of black and odorous water reduce black and odorous water
COD。
In another specific embodiment of the present invention, in the special degradation bacteria strains degraded black and odorous water of black and odorous water
Ammonia nitrogen and/or remove phosphate.
In another specific embodiment of the present invention, the black and odorous water is from river course, swag, irrigation canals and ditches.
In another specific embodiment of the present invention, the pH of black and odorous water is 6.5-8.5, preferably 7.5-8.2；It is black
The temperature of smelly water body is 10-40 DEG C, preferably 23-30 DEG C；The state of black and odorous water is preferably shaken to stand, shaking or be aerated
Swing or be aerated.
Present invention also offers a kind of microbial bacterial agent for administering black and odorous water, it includes the foregoing special drop of black and odorous water
Solve bacterium.
The special degradation bacteria strains of black and odorous water provided by the invention, have strong selectivity, energy to the pollutant in black and odorous water
Enough organic matters and ammonia nitrogen efficiently utilized in black and odorous water, the COD of black and odorous water can reduce by more than 90%, be provided simultaneously with good
Phosphorus ligands ability, black and odorous water purification of water quality efficiency can be greatly improved.
Embodiment
Embodiment 1：Separation, screening and the identification of bacterial strain
Sludge is taken from Jizhuangzi Waste Water Treatment Plant, Tianjin, being placed in sterilization container at once after sample collection takes back；Sterile bar
5g sludge is taken to shake expansion culture 24h in LB fluid nutrient mediums under part；Take 5mL nutrient solutions to transfer to train in LB fluid nutrient mediums
Support 24h (being repeated 3 times)；Last time LB nutrient solutions 1mL is taken to dilute 103-108Times, dilution is coated on LB solid mediums,
37 DEG C incubated, line separation repeatedly occurs after bacterium colony to there is pure single bacterium colony.
Another kind separation, screening mode：Sludge is taken from Jizhuangzi Waste Water Treatment Plant, Tianjin, is placed at once after sample collection
Sterilization container is taken back；5g sludge is taken to shake expansion culture 24h in LB fluid nutrient mediums under aseptic condition；5mL nutrient solutions are taken to turn
It is connected in LB fluid nutrient mediums and cultivates 24h (being repeated 3 times)；Last time LB nutrient solutions 1mL is taken to dilute 103-108Times, by dilution
It is coated on No. 1 solid medium of Gao Shi；30 DEG C incubated, line separation repeatedly occurs after bacterium colony to there is pure single bacterium colony..
10 pure single bacterium colonies of picking move to the 4 DEG C of preservations of LB slant mediums respectively.
10 bacterial strains are inoculated in ammonia nitrogen enrichment LB plating mediums (glucose 5g, (NH respectively4)2SO42g, NaCl2g,
FeSO4·7H2O 0.4g, K2HPO41g, MgSO4·7H2O 0.5g, pH 7.2) on, 37 DEG C of quiescent cultures 3 days, wherein AW0F5
Survival.
10 bacterial strains are inoculated on phosphorus enrichment LB plating mediums respectively (in per 1000mL water, glucose 10g,
(NH4)2SO40.5g, MgSO4·H2O 0.3g, NaCl0.3g, KCl0.3g, FeSO47H2O 0.03g, MnSO4·H2O
0.03g, K2HPO40.5g, pH 7.2), 37 DEG C of quiescent cultures 5 days, wherein AW0F5 is survived and Soluble phosphorus circle occurs.
The 4 DEG C of preservations of LB slant mediums will be moved to after AW0F5 bacterial strains.
The aobvious positive of strains A W0F5 Gram's staining, during using LB culture mediums, the condition of Strain survival and growth and breeding is temperature
Spend 5-55 DEG C of temperature, pH 6.5-8.0.Bacterium colony is rounded during flat board culture, and surface is smooth slightly convex, regular edges, color
For opaque milky, single thalline is into shaft-like.LB medium cultures after 24 hours thalline it is long 3.0-4.0 μm.It is general using 16S
Primer 2 7F and 1492R carry out 16S bacterium conserved sequences PCR amplification, obtain rDNA sequences and carry out BLAST sequence analysis, find with
At least 15 plants of bacillus megateriums have 97% homology (being sequenced by Beijing six directions Hua Da Gene Tech. Company Limited), can be preliminary
It is accredited as bacillus megaterium (Bacillus megaterium).AW0F5 16S rDNA sequences are as shown in SEQ ID NO.1.
SEQIDNO.1：
Embodiment 2：Ammonia nitrogen degradation
Make ammonia nitrogen liquid culture medium：Glucose 1g, (NH4)2SO40.4g, NaCl2g, FeSO4·7H2O0.4g,
K2HPO41g, MgSO4·7H2O 0.5g, 1000mL deionized waters, 7.2,121 DEG C of sterilizing 20min of pH.
AW0F5 strains are connected to LB fluid nutrient mediums by slant medium, 110r/min is cultivated 24 hours and is made at 30 DEG C
Seed liquor.2000r/min centrifugations 7min collects thalline.Bacteria suspension is made in thalline with 3mL sterilized waters, takes 0.5mL to be inoculated in ammonia
In nitrogen liquid culture medium, the blank group for not being inoculated with thalline is set, cultivated at 23 DEG C under the conditions of 110r/min.Determine within every 24 hours
Ammonia nitrogen concentration, calculate ammonia nitrogen degradation rate.
(NH in the ammonia nitrogen liquid culture medium4)2SO4(in this application be also referred to as ammonia nitrogen) is only nitrogen source, at the beginning of its ammonia nitrogen
Beginning concentration 85.5mg/L, by 1 day, 7 days and 10 days, substrate ammonia nitrogen degradation rate reached 55.2%, 63.9% and 85.6%.
Ammonia nitrogen degradation rate calculation formula：
Wherein, X is ammonia nitrogen degradation rate；M1 is experimental group ammonia nitrogen concentration (mg/L)；M2 is blank group ammonia nitrogen concentration (mg/L)；
M is initial ammonia nitrogen concentration (mg/L).
Embodiment 3：The removal of phosphorus
Make ammonia nitrogen liquid culture medium：CH3COONa 1g, MgSO4·7H2O 0.082g, FeSO4·7H2O0.0037g,
CaCl20.06g, (NH4)2SO40.2g, beef extract 0.22g, K2HPO40.057g, pH7.2,121 DEG C of sterilizing 20min.
Strain AW0F5 is connected to LB fluid nutrient mediums by slant medium, 110r/min is cultivated 24 hours and is made at 30 DEG C
Seed liquor.2000r/min centrifugations 7min collects thalline.Bacteria suspension is made in thalline with 3mL sterilized waters, takes 0.5mL to be inoculated in ammonia
In nitrogen liquid culture medium, the blank group for not being inoculated with thalline is set, cultivated at 23 DEG C under the conditions of 110r/min.Determine within every 24 hours
Phosphate concentration, calculate dephosphorizing rate.
Initial phosphate concentration is determined as 23.75mg/L, and 7 days dephosphorizing rates are 57.9% after inoculation.
Dephosphorizing rate calculation formula：
Wherein, Y is dephosphorizing rate rate；N1 is experimental group phosphate concentration (mg/L)；N2 is blank group phosphate concentration (mg/
L)；N is initial phosphate concentration (mg/L).
Embodiment 4：Black and odorous water raw water is degraded
Black and odorous water used in the present embodiment is the water body of Tianjin Beichen District green light town swag；After measured, black smelly water
The COD of body raw waterCrIt is 240mg/L；Ammonia-nitrogen content is 50mg/L；Phosphorus content is 13mg/L.
Strain AW0F5 is connected to LB fluid nutrient mediums by slant medium, 110r/min is cultivated 24 hours and is made at 30 DEG C
Seed liquor.Thalline is collected by centrifugation.
Bacteria suspension is made in thalline with 5mL sterilized waters, takes 1mL to add into black and odorous water raw water, the 50r/min at 23 DEG C
Under the conditions of place.The COD of every 24 hours measure black and odorous water raw waters, and ammonia nitrogen and total phosphorus concentration, calculate COD reduced rates, ammonia
Nitrogen degradation rate and dephosphorizing rate, as a result as shown in table 1.
Each index content in water body after the processing different number of days of table 1
| CODCr/mg/L | Ammonia-nitrogen content/mg/L | Phosphorus content/mg/L | |
| 6h | 231.42 | 49.35 | 12.28 |
| 12h | 220.65 | 48.89 | 11.65 |
| 1 day | 196.81 | 47.05 | 10.58 |
| 3 days | 76.32 | 39.22 | 6.21 |
| 7 days | 23.52 | 24.93 | 5.10 |
| 15 days | 19.68 | 2.25 | 5.50 |
CODCrRefer to that with potassium bichromate be oxygen demand that oxidant is measured, be the value that COD is measured with potassium dichromate method.
From table 1,3 days, 7 days and 15 days COD reduce 68.2%, 90.2% and 91.8% respectively after inoculation；Bacterial strain
10 days and 15 days ammonia nitrogen degradation rates are respectively 54.4% and 95.5% after inoculation；7 days dephosphorizing rates reach 60.8% after inoculation.
A kind of special degradation bacteria of black and odorous water provided by the present invention and its application are described in detail above.Herein
In apply specific embodiment to the present invention principle and embodiment be set forth, the explanation of above example is only intended to
Help to understand method and its central idea of the invention.It should be pointed out that for the person of ordinary skill of the art, do not taking off
On the premise of from the principle of the invention, some improvement and modification can also be carried out to the present invention, these are improved and modification also falls into this
The protection of invention claim.
Claims (9)
1. a kind of special degradation bacteria strains of black and odorous water, it is characterised in that the bacterial strain is bacillus megaterium, deposit number
CGMCC No.13543。
A kind of 2. method for expanding the special degradation bacteria strains of black and odorous water described in claim 1, it is characterised in that use LB liquid
Culture medium；Cultivation temperature is 5-55 DEG C, preferably 30-40 DEG C；PH is 6.8-8.0, preferably 7.2-7.8.
A kind of 3. method for expanding the special degradation bacteria strains of black and odorous water described in claim 1, it is characterised in that use Gao Shi 1
Number fluid nutrient medium；Cultivation temperature is 15-45 DEG C, preferably 25-40 DEG C；PH is 6.8-7.5.
4. application of the special degradation bacteria strains of black and odorous water according to claim 1 in black and odorous water is administered.
5. application according to claim 2, it is characterised in that the special degradation bacteria strains of black and odorous water reduce black and odorous water
COD.
6. application according to claim 2, it is characterised in that the special degradation bacteria strains degraded black and odorous water of black and odorous water
In ammonia nitrogen and/or remove phosphate.
7. application according to claim 2, it is characterised in that the black and odorous water comes from river course, swag or irrigation canals and ditches.
8. according to the application any one of claim 4-7, it is characterised in that the pH of black and odorous water is 6.5-8.5, preferably
For 7.5-8.2；The temperature of black and odorous water is 10-40 DEG C, preferably 23-30 DEG C；The state of black and odorous water is standing, shakes or expose
Gas, preferably shake or be aerated.
9. a kind of microbial bacterial agent for administering black and odorous water, it is characterised in that special comprising the black and odorous water described in claim 1
Degradation bacteria.
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Cited By (5)
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| CN109402029A (en) * | 2018-12-20 | 2019-03-01 | 北京本农科技发展有限公司 | Isolation and purification method, ammonia nitrogen degradation strain and the application of ammonia nitrogen degradation bacterium |
| CN109455823A (en) * | 2018-12-04 | 2019-03-12 | 扬州红硕环境与生物工程研究有限公司 | Phosphatic preparation and its preparation method and application in a kind of fixed water body |
| CN110157631A (en) * | 2018-03-27 | 2019-08-23 | 新疆水处理工程技术研究中心有限公司 | A kind of heat-resisting dephosphorization composite functional bacterial agent and preparation method thereof |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110157631A (en) * | 2018-03-27 | 2019-08-23 | 新疆水处理工程技术研究中心有限公司 | A kind of heat-resisting dephosphorization composite functional bacterial agent and preparation method thereof |
| CN108977398A (en) * | 2018-09-13 | 2018-12-11 | 成都友益佳环保设备工程有限公司 | One plant of bacillus megaterium and its application |
| CN108977398B (en) * | 2018-09-13 | 2021-10-08 | 成都友益佳环保设备工程有限公司 | Bacillus megaterium and application thereof |
| CN109455823A (en) * | 2018-12-04 | 2019-03-12 | 扬州红硕环境与生物工程研究有限公司 | Phosphatic preparation and its preparation method and application in a kind of fixed water body |
| CN109402029A (en) * | 2018-12-20 | 2019-03-01 | 北京本农科技发展有限公司 | Isolation and purification method, ammonia nitrogen degradation strain and the application of ammonia nitrogen degradation bacterium |
| CN109402029B (en) * | 2018-12-20 | 2020-11-10 | 北京本农科技发展有限公司 | Separation and purification method of ammonia nitrogen degrading bacteria, ammonia nitrogen degrading bacteria and application |
| CN116463232A (en) * | 2022-01-11 | 2023-07-21 | 河北省科学院生物研究所 | Glutamine bacillus G2 and application thereof |
| CN116463232B (en) * | 2022-01-11 | 2023-09-19 | 河北省科学院生物研究所 | Glutamine bacillus G2 and application thereof |
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