CN106399176A - Paenibacillus and its application in water body purification - Google Patents
Paenibacillus and its application in water body purification Download PDFInfo
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- CN106399176A CN106399176A CN201610875833.1A CN201610875833A CN106399176A CN 106399176 A CN106399176 A CN 106399176A CN 201610875833 A CN201610875833 A CN 201610875833A CN 106399176 A CN106399176 A CN 106399176A
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- 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
- C12N1/205—Bacterial isolates
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
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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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- 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
- C02F2101/30—Organic compounds
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Abstract
The invention relates to a water body purifying microorganism, and particularly discloses a paenibacillus YE-1 and its application. The paenibacillus YE-1 is preserved at China General Microbiological Culture Collection Center, the preservation date is August 4, 2016 and the preservation number is CGMCC No. 12814. The paenibacillus YE-1 can be used for effectively absorbing degrading organic matters in black and odorous waste in a biological oxidation reactor, and purifying water quality. Furthermore, since a distributed photovoltaic system supplies power energy to an aeration device of the biological oxidation reactor, the technical flow needs not additional power energy, the operation cost is low, and the treatment is effective and stable.
Description
Technical field
The present invention relates to water body purification microorganism, specifically, it is related to Bacillus species and its application.
Background technology
With the quickening of economic fast development and urbanization, China's many urban river water quality pollutions and ecological degradation are asked
Topic is very prominent, or even occurs in that seasonality and perennially black and odorous water phenomenon.Existing black and odorous water administration way mainly adopts
Based on plant purification and aeration improvement, purification load is low, and governance process needs additional electrical energy to put into, and operation cost is high.
Bacillus cereuss are the dominant populations in soil, can consumingly decompose carbon system, nitrogen system, phosphorus system, sulfur system pollutant, and energy
Decomposing protein and complicated polysaccharide, also play the role of important to the decomposition of water soluble organic substance, simultaneously can with breeding environment in
Harmful algae and aquatic pathogenic bacteria competition, form dominant population, suppression harmful algae and aquatic pathogenic bacteria.Characteristic due to it
It is better than photosynthetic bacteria with function and is expected to become the succedaneum of photosynthetic bacteria, it has also become the currently focus class of international water purification circle research
Topic.
Publication No. is that the Chinese patent of CN103923868B discloses a kind of mixed bacterial microorganism formulation, is by class spore
Bacillus, the bacteria suspension of Pseudomonas stutzeri, pseudomonas pseudoalcaligenes and Pseudomonas oleovorans composition, total viable count is 3.8*
108More than CFU/ml.Described mixed bacterial microorganism formulation can be used for processing sewage, can strengthen autumn artificial wetland treatment
Polluted river water and the ability of town domestic sewage.However, described microorganism formulation relies on the composition of multiple-microorganism and plays
Function, its need above-mentioned multiple-microorganism is carried out to separate, screen and enrichment culture after be prepared, workload is big, and cost
Higher.
Content of the invention
In order to solve problems of the prior art, it is an object of the invention to provide Bacillus species and its net
Change the application of water body aspect.
In order to realize the object of the invention, technical scheme is as follows：
In a first aspect, the invention provides Bacillus species, being identified as bacillus genus
(Paenibacillus sp.), numbering YE-1, it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center
(abbreviation CGMCC, address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, postcode：
100101), preservation date is August in 2016 4, and deposit number is CGMCC No.12814.
This bacterial strain have active adsorption, degraded black and odorous water in Organic substance function.
Invention further provides application in purifying water body for the described series bacillus YE-1.
Second aspect, the invention provides a kind of microbial bacterial agent for purifying water body, containing described series bacillus
YE-1.Further, in described microbial inoculum, the quantity of series bacillus YE-1 is not less than 108Individual/mL.
The third aspect, the invention provides a kind of method of purifying water body, methods described comprises the steps：
S1, water body to be clean is passed through biooxidation reactions device；
S2, by after described series bacillus YE-1 enrichment culture, be seeded to biooxidation reactions device；
S3, biochemical reactor, with room temperature-operating, are passed through air and carry out Air Exposure, hydraulic retention 2～4 hours；At this
During individual, series bacillus can in biochemical reactor flourish, and complete water body organism degradation process；
S4, biooxidation reactions device water outlet enter solid-liquid separation reactor, and natural subsidence completes water outlet after solid-liquid separation, obtains
To purifying water body.
Further, in described biooxidation reactions device, filler is commercially available silicon substrate porous aggregate.
Preferably, in S2, the addition of series bacillus YE-1 is not less than 2 × 10 with respect to water body to be clean11～4 ×
1011Individual/m3.
Further, in S2, the method for described enrichment culture is to inoculate described series bacillus YE-1 into enrichment culture
In base, under 25～30 DEG C of cultivation temperature, 100rpm shaking table culture reaches 10 to bacterial concentration8Individual/mL.
Preferably, the formula of described enrichment medium is：0.5g/L yeast extract, 0.5g/L glucose, 0.1g/L
Calcium chloride, 0.05g/L dipotassium hydrogen phosphate, 0.5g/L ammonium sulphate, 1000mL distilled water, pH=7.0.During configuration, each composition is mixed
Uniformly, adjust pH=7.0, sterilize 25 minutes at 121 DEG C.
Described enrichment medium can be additionally used in separation and Culture series bacillus as described above, during as isolation medium,
30g/L gellan gum will be added after composition mix homogeneously in formula during configuration, adjust pH=7.0, sterilizing at 121 DEG C was down flat after 25 minutes
Plate.
Preferably, the method for the invention is to COD>40mg/L, total nitrogen>15mg/L, total phosphorus>The easy anoxia of 1mg/L is black
Smelly water body has preferably clean-up effect.
Further, when processing high-concentration difficult-degradation organic sewage using series bacillus YE-1 of the present invention, can
With to complex biological nutritional preparation in handled sewage, to meet the growth needs of the strain of the present invention.
Further. the electric energy of the aeration facility in reactor is supplied as distributed photovoltaic system.With distributed photovoltaic
The aerating system being used in conjunction can provide suitable growing environment for this strain and be that this strain degradation Organic substance supplies oxygen, above-mentioned
Technological process need not be additionally provided electric energy, and to black and odorous water organic matter degradation efficiency high, operation cost is low, processes effectively and steady
Fixed.
Series bacillus YE-1 of the present invention has the spy such as good to city black and odorous water strong adaptability, biological stability
Point.They not only can be effectively reduced the COD of sewage, its sharpest edges be can low temperature (<5 DEG C) under the conditions of sewage is entered
Row is processed, and still has good microbial activity.In one embodiment of the invention, carry out biochemical scene using this strain
Experiment, plant running 180 days, growth of microorganism is good, system stable water outlet.
The beneficial effects of the present invention is：
The present invention newly isolates Bacillus species, can effective absorption degradation black and odorous water Organic substance, purify water.
Additionally, the aerating system being used in conjunction with distributed photovoltaic can be had for the suitable growing environment of this strain offer and for this strain degradation
Machine thing supplies oxygen, and above-mentioned technological process need not be additionally provided electric energy, to black and odorous water organic matter degradation efficiency high, operation cost
Low, process effectively and stable.
Brief description
Fig. 1 is the stereoscan photograph of series bacillus YE-1 of the present invention.
Fig. 2 is the distributed photovoltaic electrical schematic described in the embodiment of the present invention 2.
Specific embodiment
Below in conjunction with embodiment, the preferred embodiment of the present invention is described in detail.It will be appreciated that it is following real
Applying providing merely to playing descriptive purpose of example, being not used to the scope of the present invention is limited.The skill of this area
Art personnel, in the case of without departing substantially from spirit of the invention and spirit, can carry out various modifications and replace to the present invention.
Experimental technique used in following embodiments if no special instructions, is conventional method.
Material used, reagent etc. in following embodiments, if no special instructions, all commercially obtain.
For the above-mentioned series bacillus of separation and Culture culture medium (referred to as：Isolation medium), formula is：
0.5g/L yeast extract, 0.5g/L glucose, 0.1g/L calcium chloride, 0.05g/L dipotassium hydrogen phosphate, 0.5g/L sulfur
Sour ammonia, 1000mL distilled water, gellan gum 30g/L.
Gellan gum will be added after composition mix homogeneously beyond gellan gum during configuration, adjust pH7.0, sterilize 25 points at 121 DEG C
Clock.Configure 100ml culture medium according to the above ratio, every 20mL falls one flat plate.
For the above-mentioned series bacillus of enrichment culture culture medium (referred to as：Enrichment medium), formula is：0.5g/L yeast
Extract, 0.5g/L glucose, 0.1g/L calcium chloride, 0.05g/L dipotassium hydrogen phosphate, 0.5g/L ammonium sulphate, 1000mL distilled water.
By each composition mix homogeneously during configuration, adjust pH7.0, sterilize 25 minutes at 121 DEG C.
The screening of embodiment 1 bacterial strain and identification
1st, screen
1) contain in described series bacillus mud sample in 100mL and add 0.3g yeast powder, gently shake in 30 DEG C of shaking table 100rpm
Swing enrichment culture 1 week, judge bacterial growth situation using microscopic examination；Guangxi Long Jianghe taken from by mud sample.Bacterial growth is to survey
Amount light absorption value at 600nm for the inoculum, if the numerical value of the OD600 obtaining is between 0.6～0.8.
2) adopt membrane filtration by the bacteriological filter in above-mentioned culture fluid, and by antibacterial with the aseptic washing of 20mL.With 5%
Inoculum concentration is inoculated in multiple 100mL enrichment mediums respectively, and 45 DEG C of degree are cultivated, and arranges and do not inoculate comparison (CK).
100rpm shaking table culture observes bacterial growth situation after two weeks.
3) enrichment culture base fluid gradient dilution 1,2,3,4,5 (is corresponded to 10 respectively-1, 10-2, 10-3, 10-4, 10-5), respectively
Take 100 μ L diluents to be coated on the flat board of isolation medium preparation, cultivate three days for 45 DEG C.Choose single bacterium colony and carry out further flat board
Line, separates and obtains single bacterium colony.After picking single bacterium colony, proceed in new solid medium flat board, using plate streaking partition method
Proceed separation and Culture, until obtaining single bacterium colony.
2nd, identify
Observed by strain morphology and physical and chemical index analysis, pick out 12 plants of bacterial strains and carry out testing at indoor black and odorous water
Reason experiment, experiment water body is 1000mL, in before processing water ammonia nitrogen be 15 ± 2mg/L, total nitrogen be 50 ± 2mg/L and COD for 300 ±
3mg/L, it is 2 × 10 that experiment connects bacterium amount8Individual, 25 DEG C of shaking table cultures 15h, the such as table 1 of the water quality index after process.
1 12 plants of isolated strains of table process water quality parameter (unit before and after black and odorous water：mg/L)
As can be seen from the above table, numbering be YE-1 bacterial strain process sewage resultant effect optimal.Process through YE-1 bacterial strain
After 15h, the clearance of chemical oxygen demand in sewage (COD), total nitrogen (TN) and total phosphorus (P) respectively reaches 80%, 80% and 70%,
The ability of its purifying water body is the strongest.
The somatic cells of YE-1 bacterial strain are in rod-short, and size is 1 μm of 2 μ m.Gram-positive, produces spore, and spore is circle
Shape or ellipse, middle life or closely middle life, size is 1 μm.Bacterium colony is circular or sub-circular, and rough surface is flat, and slightly gloss is (seemingly
Wax candle sample color), colony diameter 1～2mm (as shown in Figure 1).
Identify strain YE-1 with 16S rDNA clone library technical Analysis.The centrifugation of 1mL bacterium solution is obtained bacterium mud, extracts total
DNA, expands 16S rDNA fragment using round pcr with prokaryote universal primer 530f and 1490r.PCR primer after purification with
The T-easy carrier of Promega connects, and converts bacillus coli DH 5 alpha.The white colony of picking determines positive bacteria by bacterium colony PCR
Fall, through enzyme action typing, to 4 cloning and sequencings.Gained sequence relatively shows through Blast, and this bacterial strain belongs to thin for Paenibacillus
Bacterium, is named as Paenibacillus YE-1.
Embodiment 2 purifying water body
1st, typical black and odorous water (the cured mountains and rivers in Jinan black and odorous water) is passed through biooxidation reactions device, this biooxidation reactions
Commercially available silicon substrate porous aggregate is filled in device.
This waste water quality is as follows：pH7.0；COD (COD), 45mg/L；Total nitrogen (TN), 15mg/L, total phosphorus (P),
3mg/L.
2nd, by the strain of identified series bacillus, it is seeded in 25 DEG C of shaking table 100rpm in enrichment medium and cultivates to bacterium
Concentration is 108Individual/mL；Then bacterium solution (also referred to as microbial inoculum, with microbial inoculum described in content of the invention) is seeded to biological oxidation anti-
Answer in device, inoculum concentration is 3L/m3.Air is filled in biooxidation reactions device bottom, aeration rate different, hydraulic retention according to water quality difference
1 hour；
3rd, the electric energy of aerator is supplied as distributed photovoltaic system；
4th, water outlet after reacting is passed through solid-liquid separation reactor, staticly settles water outlet after solid-liquid separation；
5th, whole response system continuously runs 30 days, and effluent quality is analyzed as follows：COD (COD) clearance
70%, total nitrogen (TN) clearance 80%, total phosphorus (P) clearance 70%.
The method of the process black and odorous water that the present invention provides, being powered by distributed photovoltaic provides aeration (as shown in Figure 2),
Under the absorption degradation of series bacillus strain acts on, black and odorous water can obtain effective process.
Purifying water body under embodiment 3 low temperature
1st, typical black and odorous water (the cured mountains and rivers in Jinan black and odorous water) is passed through biooxidation reactions device, this biooxidation reactions
Commercially available silicon substrate porous aggregate is filled, this waste water quality is as follows in device：pH7.0；COD (COD), 45mg/L；Total nitrogen
(TN), 15mg/L, total phosphorus (P), 3mg/L.
2nd, by the strain of identified series bacillus, it is seeded in enrichment medium, 5 DEG C of shaking table 100rpm cultivate to bacterium
Concentration is 108Individual/mL；Then by bacterium solution (also referred to as microbial inoculum, with microbial inoculum described in content of the invention) be seeded to low temperature environment (<5
DEG C) in biooxidation reactions device, inoculum concentration is 3L/m3.Air is filled in biooxidation reactions device bottom, and aeration rate is according to water quality difference
Difference, hydraulic retention 1 hour；
3rd, the electric energy of aerator is supplied as distributed photovoltaic system；
4th, water outlet after reacting is passed through solid-liquid separation reactor, staticly settles water outlet after solid-liquid separation；
5th, whole response system continuously runs 30 days, and effluent quality is analyzed as follows：COD (COD) clearance
67%, total nitrogen (TN) clearance 78%, total phosphorus (P) clearance 60%.Can be seen that this bacterial strain still has at low temperature stronger
Black and odorous water detergent power.
Although, above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to the scope of protection of present invention.
Claims (10)
1. a Bacillus species YE-1, its deposit number is CGMCC No.12814.
2. application in purifying water body for the series bacillus YE-1 described in claim 1.
3. a kind of microbial bacterial agent for purifying water body is it is characterised in that it contains the series bacillus described in claim 1
YE-1.
4. microbial bacterial agent according to claim 3 it is characterised in that in described microbial inoculum series bacillus YE-1 quantity
Not less than 108Individual/mL.
5. a kind of method of purifying water body is it is characterised in that methods described comprises the steps：
S1, water body to be clean is passed through biooxidation reactions device；
S2, by after the series bacillus YE-1 enrichment culture described in claim 1, be seeded to biooxidation reactions device；
S3, biochemical reactor, with room temperature-operating, are passed through air and carry out Air Exposure, hydraulic retention 2～4 hours；
S4, biooxidation reactions device water outlet enter solid-liquid separation reactor, and natural subsidence completes water outlet after solid-liquid separation, obtain net
Change water body.
6. method according to claim 5 is it is characterised in that in S2, and the addition of series bacillus YE-1 is with respect to treating
Purifying water body is not less than 2 × 1011～4 × 1011Individual/L.
7. method according to claim 5 is it is characterised in that in S2, the method for described enrichment culture is by described class bud
Spore bacillus YE-1 inoculates in enrichment medium, and under 25～30 DEG C of cultivation temperature, shaking table culture reaches 10 to bacterial concentration8
Individual/mL.
8. method according to claim 7 is it is characterised in that the formula of described enrichment medium is：0.5g/L yeast soaks
Extract, 0.5g/L glucose, 0.1g/L calcium chloride, 0.05g/L dipotassium hydrogen phosphate, 0.5g/L ammonium sulphate, 1000mL distilled water, pH
=7.0.
9. the method according to any one of claim 5～8 is it is characterised in that described water body to be clean is COD>40mg/L、
Total nitrogen>15mg/L, total phosphorus>The black smelly water body of easy anoxia of 1mg/L.
10. method according to claim 8 is it is characterised in that the electric energy of aeration facility is supplied as distributed photovoltaic system.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108865920A (en) * | 2017-05-11 | 2018-11-23 | 北京有色金属研究总院 | One plant of resistance to cadmium, the organic matter degradation bacteria of lead ion and its application |
| CN110643554A (en) * | 2019-11-26 | 2020-01-03 | 中国科学院烟台海岸带研究所 | Strain capable of efficiently removing inorganic phosphorus in water body and application thereof |
| CN111172076A (en) * | 2019-12-17 | 2020-05-19 | 中国科学院上海高等研究院 | Bacterial strain for treating feces in ecological toilet and application thereof |
| CN111269855A (en) * | 2020-02-19 | 2020-06-12 | 常州大学 | Algicidal bacteria, screening method and application |
| CN112553188A (en) * | 2020-10-27 | 2021-03-26 | 杭州秀川科技有限公司 | Carbon source slow-release wax loaded with microorganisms as well as preparation method and application thereof |
| CN113583924A (en) * | 2021-10-08 | 2021-11-02 | 佛山市玉凰生态环境科技有限公司 | Riverway water body COD degrading strain and application thereof |
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| CN112553188A (en) * | 2020-10-27 | 2021-03-26 | 杭州秀川科技有限公司 | Carbon source slow-release wax loaded with microorganisms as well as preparation method and application thereof |
| CN113583924A (en) * | 2021-10-08 | 2021-11-02 | 佛山市玉凰生态环境科技有限公司 | Riverway water body COD degrading strain and application thereof |
| CN113583924B (en) * | 2021-10-08 | 2022-02-01 | 佛山市玉凰生态环境科技有限公司 | Riverway water body COD degrading strain and application thereof |
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| CN106399176B (en) | 2019-08-13 |
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