CN106399176B - One Bacillus species and its application in terms of purifying water body - Google Patents
One Bacillus species and its application in terms of purifying water body Download PDFInfo
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- CN106399176B CN106399176B CN201610875833.1A CN201610875833A CN106399176B CN 106399176 B CN106399176 B CN 106399176B CN 201610875833 A CN201610875833 A CN 201610875833A CN 106399176 B CN106399176 B CN 106399176B
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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
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- 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 present invention relates to water body purification microorganisms, specifically disclose a Bacillus species YE-1 and its application.The series bacillus YE-1, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and the deposit date is on August 4th, 2016, deposit number was CGMCC No.12814.Using series bacillus YE-1 of the present invention, in biooxidation reactions device can effective absorption degradation black and odorous water organic matter, purify water.Further, electric energy is supplied by the aeration facility that distributed photovoltaic system is biological oxidation reactor, without being additionally provided electric energy, operation cost is low for process flow, handles effectively and stablizes.
Description
Technical field
The present invention relates to water body purification microorganisms, specifically, being related to a Bacillus species and its application.
Background technique
With the quickening of economic fast development and urbanization, many urban river water quality pollutions in China and ecological degradation are asked
Topic is very prominent, or even seasonality and perennially black and odorous water phenomenon occurs.Existing black and odorous water administration way mainly uses
Based on plant purification and aeration are administered, purification load is low, and governance process needs additional electrical energy to put into, and operation cost is high.
Bacillus is the dominant population in soil, can consumingly decompose carbon system, nitrogen system, phosphorus system, sulphur system pollutant, and energy
Decomposing protein and complicated polysaccharide also have an important role to the decomposition of water soluble organic substance, at the same can in breeding environment
Harmful algae and aquatic pathogenic bacteria competition, form dominant population, inhibit harmful algae and aquatic pathogenic bacteria.Due to its characteristic
It is expected to become the substitute of photosynthetic bacteria better than photosynthetic bacteria with function, it has also become the hot spot class of current world 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 gemma
The bacteria suspension of bacillus, Pseudomonas stutzeri, pseudomonas pseudoalcaligenes and Pseudomonas oleovorans composition, total viable count are 3.8*
108CFU/ml or more.The mixed bacterial microorganism formulation can be used for handling sewage, can reinforce autumn artificial wetland treatment
The ability of polluted river water and town domestic sewage.However, the microorganism formulation relies on the composition of multiple-microorganism and plays
Function needs that above-mentioned multiple-microorganism is separated, screens and prepared after enrichment culture, heavy workload, and cost
It is higher.
Summary of the invention
In order to solve the problems in the existing technology, the object of the present invention is to provide a Bacillus species and its net
Change the application in terms of water body.
In order to achieve the object of the present invention, technical scheme is as follows:
In a first aspect, being identified as bacillus genus the present invention provides a Bacillus species
(Paenibacillus sp.), number YE-1 are 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), the deposit date is on August 4th, 2016, deposit number is CGMCC No.12814.
The bacterial strain have the function of effectively adsorb, degradation black and odorous water in organic matter.
Invention further provides application of the series bacillus YE-1 in purifying water body.
Second aspect, the present invention provides a kind of microbial bacterial agents for purifying water body, contain the series bacillus
YE-1.Further, the quantity of series bacillus YE-1 is not less than 10 in the microbial inoculum8A/mL.
The third aspect, the present invention provides a kind of methods of purifying water body, and described method includes following steps:
S1, water body to be clean is passed through biooxidation reactions device；
S2, by after the series bacillus YE-1 enrichment culture, be seeded to biooxidation reactions device；
S3, biochemical reactor are passed through air and are carried out Air Exposure with room temperature-operating, and hydraulic retention 2~4 hours；At this
During a, series bacillus can in biochemical reactor flourish, and complete water body organism degradation process；
S4, the water outlet of biooxidation reactions device, which enter, is separated by solid-liquid separation reactor, and natural subsidence is completed to be discharged after being separated by solid-liquid separation, and obtains
To purifying water body.
Further, filler is commercially available silicon substrate porous aggregate in the biooxidation reactions device.
Preferably, the additive amount of series bacillus YE-1 is not less than 2 × 10 relative to water body to be clean in S211~4 ×
1011A/m3。
Further, in S2, the method for the enrichment culture is that the series bacillus YE-1 is inoculated with into enrichment culture
In base, under 25~30 DEG C of cultivation temperature, 100rpm shaking table culture to bacterial concentration reaches 108A/mL.
Preferably, the formula of the enriched medium are as follows: 0.5g/L yeast extract, 0.5g/L glucose, 0.1g/L
Calcium chloride, 0.05g/L dipotassium hydrogen phosphate, 0.5g/L sulfate of ammoniac, 1000mL distilled water, pH=7.0.Each ingredient is mixed when configuration
Uniformly, pH=7.0 is adjusted, is sterilized 25 minutes at 121 DEG C.
The enriched medium can also be used to be separately cultured series bacillus as described above, when as isolation medium,
When configuration 30g/L gellan gum is added in ingredient in formula after mixing, adjusts pH=7.0, fallen after sterilizing 25 minutes at 121 DEG C flat
Plate.
Preferably, the method for the invention is black to COD > 40mg/L, total nitrogen > 15mg/L, the easy anoxic of total phosphorus > 1mg/L
Smelly water body has preferable clean-up effect.
It further, can when using series bacillus YE-1 of the present invention processing high-concentration difficult-degradation organic sewage
With the complex biological nutritional preparation into handled sewage, to meet the growth needs of strain of the invention.
The electric energy of aeration facility further in reactor is supplied as distributed photovoltaic system.With distributed photovoltaic
The aerating system being used in conjunction can provide suitable growing environment for the strain and be strain degradation organic matter supply oxygen, above-mentioned
Process flow is without being additionally provided electric energy, and high-efficient to black and odorous water organic matter degradation, operation cost is low, and processing is effectively and steady
It is fixed.
Series bacillus YE-1 of the present invention has the spies such as adaptable to city black and odorous water, biological stability is good
Point.They can not only be effectively reduced the COD of sewage, sharpest edges be can under the conditions of low temperature (< 5 DEG C) to sewage into
Row processing, still has good microbial activity.In one embodiment of the invention, biochemical scene is carried out using the strain
Experiment, device are run 180 days, microorganism well-grown, system stable water outlet.
The beneficial effects of the present invention are:
The present invention newly isolates a Bacillus species, can effective absorption degradation black and odorous water organic matter, purify water.
In addition, the aerating system being used in conjunction with distributed photovoltaic can provide suitable growing environment for the strain and be that the strain degradation has
Machine object supplies oxygen, and above-mentioned process flow is without being additionally provided electric energy, operation cost high-efficient to black and odorous water organic matter degradation
Low, processing is effectively and stable.
Detailed description of the invention
Fig. 1 is the stereoscan photograph of series bacillus YE-1 of the present invention.
Fig. 2 is distributed photovoltaic electrical schematic described in the embodiment of the present invention 2.
Specific embodiment
The preferred embodiment of the present invention is described in detail below in conjunction with embodiment.It will be appreciated that following real
Providing merely to play the purpose of explanation for example is applied, is not used to limit the scope of the present invention.The skill of this field
Art personnel without departing from the spirit and purpose of the present invention, can carry out various modifications and replace to the present invention.
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
For being separately cultured the culture medium (referred to as: isolation medium) of above-mentioned series bacillus, formula are as follows:
0.5g/L yeast extract, 0.5g/L glucose, 0.1g/L calcium chloride, 0.05g/L dipotassium hydrogen phosphate, 0.5g/L sulphur
Sour ammonia, 1000mL distilled water, gellan gum 30g/L.
When configuration gellan gum is added in the ingredient other than gellan gum after mixing, adjusts pH7.0, sterilize 25 points at 121 DEG C
Clock.100ml culture medium is configured according to the above ratio, and every 20mL falls one flat plate.
For the culture medium (referred to as: enriched medium) of the above-mentioned series bacillus of enrichment culture, formula are as follows: 0.5g/L yeast
Extract, 0.5g/L glucose, 0.1g/L calcium chloride, 0.05g/L dipotassium hydrogen phosphate, 0.5g/L sulfate of ammoniac, 1000mL distilled water.
Each ingredient is uniformly mixed when configuration, pH7.0 is adjusted, sterilizes 25 minutes at 121 DEG C.
The screening and identification of 1 bacterial strain of embodiment
1, it screens
1) it is gently shaken containing 0.3g yeast powder is added in the series bacillus mud sample in 30 DEG C of shaking table 100rpm in 100mL
It swings enrichment culture 1 week, judges bacterial growth situation using microscope detection；Mud sample is derived from Guangxi Long Jianghe.Bacterial growth is extremely surveyed
Light absorption value of the inoculum at 600nm is measured, if the numerical value of obtained OD600 is between 0.6~0.8.
2) it is filtered using film by the bacteriological filter in above-mentioned culture solution, and by bacterium under the sterile washing of 20mL.With 5%
Inoculum concentration is inoculated in respectively in multiple 100mL enriched mediums, and 45 DEG C of degree are cultivated, and is arranged and is not inoculated with control (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 respectively corresponded 10-1, 10-2, 10-3, 10-4, 10-5), respectively
100 μ L dilutions are taken to be coated on the plate of isolation medium preparation, 45 DEG C are cultivated three days.It chooses single colonie and carries out further plate
Scribing line, isolated single colonie.It after picking single colonie, is transferred in new solid medium tablets, utilizes plate streaking partition method
Continue to be separately cultured, until obtaining single colonie.
2, it identifies
By strain morphology observation and physical and chemical index analysis, picks out 12 plants of bacterial strains and test at indoor black and odorous water
Reason experiment, experiment water body are 1000mL, in the preceding water of processing ammonia nitrogen be 15 ± 2mg/L, total nitrogen be 50 ± 2mg/L and COD be 300 ±
3mg/L, it is 2 × 10 that experiment, which connects bacterium amount,8It is a, 25 DEG C of shaking table culture 15h, water quality indicator that treated such as table 1.
Water quality parameter (unit: mg/L) before and after 1 12 plants of isolated strains processing black and odorous waters of table
As can be seen from the above table, the resultant effect for the bacterial strain processing sewage that number is YE-1 is best.It is handled through YE-1 bacterial strain
After 15h, the removal rate 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 most strong.
The somatic cells of YE-1 bacterial strain are in rod-short, and size is 1 μm of 2 μ m.Gram-positive, produces gemma, and gemma is circle
Shape or ellipse, middle life or close middle raw, size is 1 μm.Bacterium colony is round or approximate circle, and rough surface is flat, and slightly gloss is (seemingly
Candle sample color), 1~2mm of colony diameter (as shown in Figure 1).
Strain YE-1 is analyzed and identified with 16S rDNA clone library technology.1mL bacterium solution is centrifuged to obtain bacterium mud, is extracted total
DNA expands 16S rDNA segment using round pcr with prokaryotes universal primer 530f and 1490r.PCR product 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
It falls, through digestion parting, to 4 cloning and sequencings.Gained sequence relatively shows that the bacterial strain is that Paenibacillus belongs to thin through Blast
Bacterium is named as Paenibacillus YE-1.
2 purifying water body of embodiment
1, typical black and odorous water (the cured mountains and rivers black and odorous water in Jinan) is passed through biooxidation reactions device, the biooxidation reactions
The commercially available silicon substrate porous aggregate of filling in device.
The waste water quality is as follows: pH7.0；COD (COD), 45mg/L；Total nitrogen (TN), 15mg/L, total phosphorus (P),
3mg/L。
2, by the strain of identified series bacillus, 25 DEG C of shaking table 100rpm in enriched medium is seeded in and are cultivated to bacterium
Concentration is 108A/mL；Then it is anti-bacterium solution (also referred to as microbial inoculum, with microbial inoculum described in summary of the invention) to be seeded to biological oxidation
It answers in device, inoculum concentration 3L/m3.Air, aeration quantity different, hydraulic retention according to water quality difference are filled in biooxidation reactions device bottom
1 hour；
3, the electric energy of aerator is supplied as distributed photovoltaic system；
4, water outlet is passed through separation of solid and liquid reactor after reacting, and is discharged after staticly settling separation of solid and liquid；
5, entire reaction system is continuously run 30 days, and effluent quality is analyzed as follows: COD (COD) removal rate
70%, total nitrogen (TN) removal rate 80%, total phosphorus (P) removal rate 70%.
The method of processing black and odorous water provided by the invention, is powered by distributed photovoltaic and provides aeration (as shown in Figure 2),
Under the absorption degradation effect of series bacillus strain, black and odorous water can get and be effectively treated.
Purifying water body under 3 low temperature of embodiment
1, typical black and odorous water (the cured mountains and rivers black and odorous water in Jinan) is passed through biooxidation reactions device, the biooxidation reactions
The commercially available silicon substrate porous aggregate of filling, the waste water quality are as follows in device: pH7.0；COD (COD), 45mg/L；Total nitrogen
(TN), 15mg/L, total phosphorus (P), 3mg/L.
2, it by the strain of identified series bacillus, is seeded in enriched medium, 5 DEG C of shaking table 100rpm are cultivated to bacterium
Concentration is 108A/mL；Then bacterium solution (also referred to as microbial inoculum, with microbial inoculum described in summary of the invention) is seeded to low temperature environment (< 5
DEG C) in biooxidation reactions device, inoculum concentration 3L/m3.Air is filled in biooxidation reactions device bottom, aeration quantity it is different according to water quality and
Difference, hydraulic retention 1 hour；
3, the electric energy of aerator is supplied as distributed photovoltaic system；
4, water outlet is passed through separation of solid and liquid reactor after reacting, and is discharged after staticly settling separation of solid and liquid；
5, entire reaction system is continuously run 30 days, and effluent quality is analyzed as follows: COD (COD) removal rate
67%, total nitrogen (TN) removal rate 78%, total phosphorus (P) removal rate 60%.It can be seen that the bacterial strain at low temperature still with 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, fall within the scope of the claimed invention without departing from theon the basis of the spirit of the present invention.
Claims (10)
1. a Bacillus species (Paenibacillus sp.) YE-1, deposit number is CGMCC No.12814.
2. application of the series bacillus YE-1 described in claim 1 in purifying water body.
3. a kind of microbial bacterial agent for purifying water body, which is characterized in that it contains series bacillus described in claim 1
YE-1。
4. microbial bacterial agent according to claim 3, which is characterized in that the quantity of series bacillus YE-1 in the microbial inoculum
Not less than 108A/mL.
5. a kind of method of purifying water body, which is characterized in that described method includes following steps:
S1, water body to be clean is passed through biooxidation reactions device；
S2, by after series bacillus YE-1 enrichment culture described in claim 1, be seeded to biooxidation reactions device；
S3, biochemical reactor are passed through air and are carried out Air Exposure with room temperature-operating, and hydraulic retention 2~4 hours；
S4, the water outlet of biooxidation reactions device, which enter, is separated by solid-liquid separation reactor, and natural subsidence is completed to be discharged after being separated by solid-liquid separation, and obtains net
Change water body.
6. according to the method described in claim 5, it is characterized in that, in S2, the additive amount of series bacillus YE-1 relative to
Purifying water body is not less than 2 × 1011~4 × 1011A/m3。
7. according to the method described in claim 5, it is characterized in that, the method for the enrichment culture is by the class bud in S2
Spore bacillus YE-1 is inoculated in enriched medium, and under 25~30 DEG C of cultivation temperature, shaking table culture to bacterial concentration reaches 108
A/mL.
8. the method according to the description of claim 7 is characterized in that the formula of the enriched medium are as follows: the leaching of 0.5g/L yeast
Extract, 0.5g/L glucose, 0.1g/L calcium chloride, 0.05g/L dipotassium hydrogen phosphate, 0.5g/L sulfate of ammoniac, 1000mL distilled water, pH
=7.0.
9. according to the described in any item methods of claim 5~8, which is characterized in that the water body to be clean be COD > 40mg/L,
The black smelly water body of total nitrogen > 15mg/L, the easy anoxic of total phosphorus > 1mg/L.
10. according to the method described in claim 8, it is characterized in that, the electric energy of aeration facility is supplied as distributed photovoltaic system.
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| CN108865920B (en) * | 2017-05-11 | 2021-06-08 | 有研资源环境技术研究院(北京)有限公司 | Cadmium and lead ion resistant organic matter degrading bacterium and application thereof |
| CN110643554B (en) * | 2019-11-26 | 2020-03-06 | 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 |
| CN111172076B (en) * | 2019-12-17 | 2020-10-16 | 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 |
| CN112553188B (en) * | 2020-10-27 | 2024-07-05 | 杭州秀川科技有限公司 | Microorganism-loaded carbon source slow-release wax and preparation method 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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| CN102220265A (en) * | 2011-05-11 | 2011-10-19 | 山东大学 | Denitrifying bacterial strain and application of the same in water body for nitrogen removal together with wetland plants |
| CN103103153A (en) * | 2013-01-29 | 2013-05-15 | 山东大学 | Anaerobic denitrification phosphorus-accumulating bacteria strain with denitrification and phosphorous removal effects and application thereof |
| CN104928209A (en) * | 2015-05-15 | 2015-09-23 | 中国科学院生态环境研究中心 | Paenibacillus and application thereof |
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| CN103103153A (en) * | 2013-01-29 | 2013-05-15 | 山东大学 | Anaerobic denitrification phosphorus-accumulating bacteria strain with denitrification and phosphorous removal effects and application thereof |
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