CN106635864B - A method of handling oil extraction waste water using YB-6 bacterial strain anaerobic degradations - Google Patents
A method of handling oil extraction waste water using YB-6 bacterial strain anaerobic degradations Download PDFInfo
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- CN106635864B CN106635864B CN201610566032.7A CN201610566032A CN106635864B CN 106635864 B CN106635864 B CN 106635864B CN 201610566032 A CN201610566032 A CN 201610566032A CN 106635864 B CN106635864 B CN 106635864B
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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
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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
- C02F3/28—Anaerobic digestion processes
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—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
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/10—Nature of the water, waste water, sewage or sludge to be treated from quarries or from mining activities
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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
- C02F2209/00—Controlling or monitoring parameters in water treatment
- C02F2209/20—Total organic carbon [TOC]
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- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
Abstract
The invention discloses a kind of methods handling oil extraction waste water using YB-6 bacterial strain anaerobic degradations.The separation of microorganism fungus kind is carried out using spread plate, isolation medium is reinforced clostridial medium, and activated sludge is selected 10 after gradient dilution‑6~10‑8The bacterium solution of concentration is coated in reinforced clostridial medium, constant-temperatureanaerobic anaerobic culture, then it is good to select growing way, the single bacterium colony to come in every shape, streak plating is respectively adopted to be inoculated in new reinforced clostridial medium, constant-temperatureanaerobic anaerobic culture, and carry out purifying culture three times, obtain YB-6 bacterial strains, it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, constant-temperatureanaerobic anaerobic culture is enriched in new reinforced clostridial medium, enrichment bacterium solution is made, it is inoculated into oil extraction waste water, the constant-temperatureanaerobic anaerobic culture 96h at 37 DEG C, complete the anaerobic degradation processing to oil extraction waste water.The method of the present invention operating process is convenient and efficient, good to the TOC degradation effects of oil extraction waste water.
Description
Technical field
The invention belongs to environment pollutant biological treatment technical fields, more particularly to a kind of to utilize YB-6 bacterial strain anaerobic degradations
The method for handling oil extraction waste water.
Background technology
Oil extraction waste water is mainly oil exploitation, storage, transport, dehydration and recycles the mixture for generating waste water in the process.
Oil extraction waste water complicated component, mainly contain the solid suspensions such as mud and silt, the oil of dissolving, the organic matter of benzene, hydrogen sulfide,
The components such as the heavy metals such as copper and mickel and sulfate, while it is high with COD contents, BOD contents are low, toxicity is high and dissolubility is oily
The features such as hardly possible removal, it is consequently belonging to used water difficult to degradate.The component of oil extraction waste water is to continue variation, this mainly with institute lard oil
Property, stage are related with the geologic feature of oil-water-layer, so it is useless how to be recovered the oil using the processing of prior art efficient stable environmental protection
Water is enterprise's focus of interest.Nowadays, the materilization freatment method of oil extraction waste water includes mainly electrochemical process, absorption method, ozone
Method, flocculence and catalysis method, but there are high energy consumption, low-yield and secondary pollution problems for materializing strategy technique, and do not meet
The requirement of national energy-saving environmental protection.In contrast, the high investment of bioanalysis efficiency is low, and treatment effect is stablized.At present at main biology
Logos includes activated sludge process, biofilm, anaerobic-aerobic process and natural constructed wetland, is paid attention to and at application biochemistry
Reason technology is the trend of domestic and international oil extraction waste water processing.
Anaerobic process can have certain capacity of resisting impact load with stable organic matter density and type in biological treatment；
Aerobic rule can thoroughly remove the organic matter in waste water.It is single although anaerobic process or aerobic method can handle waste water
One aerobic or anaerobic technique has some limitations.It is more and more in order to preferably handle the organic matter in waste water
Researcher two or more process combinations are got up, to improve the treatment effect of waste water.Oil extraction waste water in this research
Treatment plant uses ABR-SBR combination techniques, COD, petroleum-type, SS, S2-Removal rate be respectively 83-94%, 78-92%, 99% He
94%, water outlet reaches《Urban wastewater treatment firm pollutant emission standard》（GB18918-2002）First discharge standard.Therefore, originally
Application filters out highly effective petroleum degradation anaerobism bacterial strain out of ABR activated sludge tanks, is improved for structure efficient degreasing microbial inoculum in next step
The treatment effect of oil extraction waste water is of great significance.
Invention content
The object of the present invention is to provide a kind of method handling oil extraction waste water using YB-6 bacterial strain anaerobic degradations, this method energy
The practical oil extraction waste water generated of enough effective degradation treatments.
The specific steps are：
(1) spread plate is used to carry out the separation of microorganism fungus kind, isolation medium is reinforced clostridial medium, will be lived
Property sludge selects 10 after gradient dilution-6~10-8The bacterium solution of concentration is coated in reinforced clostridial medium, and will strengthen clostridium
Culture medium is placed in anaerobic culture box to be cultivated 7 days at 37 DEG C, to be selected spare.
(2) single bacterium that growing way is good, comes in every shape is selected from the spare reinforced clostridial medium to be selected obtained by step (1)
It falls, streak plating is respectively adopted and is inoculated in new reinforced clostridial medium, reinforced clostridial medium is then placed in anaerobism
It is cultivated at 37 DEG C in incubator 7 days, i.e. it is white to obtain well-grown anaerobism bacterial strain, bacterium colony in reinforced clostridial medium for screening
Color, irregular shape, surface is smooth, opaque, and edge is irregular, is named as YB-6, is then purified three times respectively to YB-6
Culture, obtains the pure YB-6 bacterial strains interfered without other miscellaneous bacterias.
(3) the YB-6 bacterial strains that step (2) obtains are enriched in new reinforced clostridial medium, are placed in anaerobic culture box
It is cultivated 7 days at 37 DEG C, enrichment bacterium solution is made.
(4) step (3) enrichment bacterium solution obtained is inoculated into oil extraction waste water, is enriched with the volume ratio of bacterium solution and oil extraction waste water
It is 1:150, then the constant-temperatureanaerobic anaerobic culture 96h at 37 DEG C, that is, complete the anaerobic degradation processing to oil extraction waste water.
The activated sludge is the sludge in the anaerobic baffled reactor of oil recovery terminal wastewater treatment plant, i.e. Anaerobic
Sludge in Bafflted Reactor, abbreviation ABR sludge.
The composition of the reinforced clostridial medium is as follows：9 ~ 11g/L of extracted beef powder, 9 ~ 11g/L of peptone, yeast extract powder 2 ~
4g/L, 0.5 ~ 1.5g/L of soluble starch, 4 ~ 6g/L of glucose, 0.5 ~ 1g/L of cysteine hydrochloride, 4 ~ 6g/L of sodium chloride, sodium acetate
The mass percent content of 2 ~ 4g/L, agar are 2%；The pH value 6.8 ± 0.2 of the reinforced clostridial medium.
The YB-6 bacterial strains are now preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation day
Phase：On 03 15th, 2016, deposit number：CGMCC No.12214, Classification And Nomenclature：Clostridium Clostridium sp.；Its thalline
Form and molecular biological characteristics：Thalline is rod-shaped, 0.4 ~ 0.6 2.5 ~ 3.2 μm of μ m, single or arrangement in pairs, raw, column in brood cell
Shape, cyst expand, Gram-positive.
The method of the present invention operating process is convenient and efficient, good and of low cost to the TOC degradation effects in oil extraction waste water,
It is easy to utilize.
Description of the drawings
Fig. 1 is the colonial morphology schematic diagram of the YB-6 bacterial strains obtained in the embodiment of the present invention.
Fig. 2 is the microscope thalli morphology schematic diagram of the YB-6 bacterial strains obtained in the embodiment of the present invention.
Fig. 3 is the sequence information of the YB-6 bacterial strains obtained in the embodiment of the present invention.
Fig. 4 is that the YB-6 bacterial strains obtained in the embodiment of the present invention develop tree graph to related kind of 16S rDNA sequential systems.
Specific implementation mode
Embodiment：
The present embodiment is with the ABR of Weizhou Island, Beihai, Guangxi CNOOC terminal wastewater treatment plant（Anaerobic Bafflted
Reactor, anaerobic baffled reactor）In activated sludge be sample specific implementation, completely show YB-6 bacterial strains screening, mirror
The application of fixed and its anaerobic degradation processing oil extraction waste water.
(1) spread plate is used to carry out the separation of microorganism fungus kind, isolation medium is reinforced clostridial medium, will be wide
The ABR activated sludge of western Weizhou Island, Beihai City CNOOC terminal wastewater treatment plant selects 10 after gradient dilution-7The bacterium solution of concentration is coated with
In reinforced clostridial medium, and reinforced clostridial medium is placed in anaerobic culture box and is cultivated 7 days at 37 DEG C, it is to be selected standby
With.
(2) single bacterium that growing way is good, comes in every shape is selected from the spare reinforced clostridial medium to be selected obtained by step (1)
It falls, streak plating is respectively adopted and is inoculated in new reinforced clostridial medium, reinforced clostridial medium is then placed in anaerobism
It is cultivated at 37 DEG C in incubator 7 days, i.e. it is white to obtain well-grown anaerobism bacterial strain, bacterium colony in reinforced clostridial medium for screening
Color, irregular shape, surface is smooth, opaque, and edge is irregular, as shown in Figure 1, be named as YB-6, then to YB-6 respectively into
Row purifying culture three times, obtains the pure YB-6 bacterial strains interfered without other miscellaneous bacterias.
(3) the YB-6 bacterial strains that step (2) obtains are enriched in new reinforced clostridial medium, are placed in anaerobic culture box
It is cultivated 7 days at 37 DEG C, enrichment bacterium solution is made.
(4) step (3) enrichment bacterium solution obtained is inoculated into oil extraction waste water, is enriched with the volume ratio of bacterium solution and oil extraction waste water
It is 1:150, then the constant-temperatureanaerobic anaerobic culture 96h at 37 DEG C, that is, complete the anaerobic degradation processing to oil extraction waste water.
The composition of the reinforced clostridial medium is as follows：Extracted beef powder 10g/L, peptone 10g/L, yeast extract powder 3g/L,
Soluble starch 0.5g/L, glucose 5g/L, cysteine hydrochloride 0.5g/L, sodium chloride 5g/L, sodium acetate 3g/L, the quality of agar
Relative content is 2%；The pH value 6.8 of the reinforced clostridial medium.
The YB-6 bacterial strains are now preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation day
Phase：On 03 15th, 2016, deposit number：CGMCC No.12214, Classification And Nomenclature：Clostridium Clostridium sp..
By the thalli morphology for the YB-6 bacterial strains that micro- sem observation the present embodiment obtains, as shown in Fig. 2, thalline is rod-shaped, 0.4
~ 0.6 2.5 ~ 3.2 μm of μ m individually or in pairs arranges, and raw in brood cell, column, cyst expands；Then Gram's staining mirror is carried out
It is fixed, after just contaminating, mordant dyeing, decolourizing, redye, glass slide is covered with coverslip, is placed in Classica E220LED binocular opticals
Microscope 100 × oil under the microscope, is identified as Gram-positive.
Bacterial strain base is carried out using Ezup pillar bacterial genomes DNA extraction agent boxes to the YB-6 bacterial strains that the present embodiment obtains
Because of the extraction of group DNA, operating procedure by specification carries out.The concentration of DNA sample passes through ultraviolet point of Quawell Q5000 ultramicron
Light photometer detects, while detecting DNA mass with 0.8% agarose gel electrophoresis, and is shown on Gel DocTM XR+ imagers
Picture.Molecular biology identification, obtained alkali are carried out using FMIC-QO01-002 bacterial 16 S rDNA detection methods to YB-6 bacterial strains
Basic sequence compares in GeneBank databases, with the result phylogenetic tree construction of comparison.Gained YB-6 bacterial strains are sent into
State's Microbiological Culture Collection administration committee common micro-organisms center carries out preservation, preservation date：On 03 15th, 2016, preservation
Number：CGMCC No.12214, Classification And Nomenclature：Clostridium Clostridium sp..The result shows that YB-6 is fusobacterium
（Clostridiumsp.）A potential novel species, sequence information and 16S rDNA sequential systems the development tree of bacterial strain are respectively such as
Shown in Fig. 3 and Fig. 4.
The present embodiment is as follows to the anaerobic degradation result of oil extraction waste water：Oil extraction waste water TOC contents are 112.30 ~ 147.10
Mg/L, COD content are 451 ~ 496 mg/L, and pH value takes 10ml to make between 7.76 ~ 7.91 after 37 DEG C of constant-temperatureanaerobic anaerobic culture 96h
To detect sample, the situation of change front and back in inoculation of TOC contents in oil extraction waste water is measured using Multi N/C3100 analyzers,
The result shows that YB-6 bacterial strains are 33.3% for the TOC degradation rates of oil extraction waste water.
SEQUENCE LISTING
<110>Guilin University of Technology, Hezhou College
<120>A method of handling oil extraction waste water using YB-6 bacterial strain anaerobic degradations
<130>
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1383
<212> DNA
<213>Fusobacterium（Clostridium sp.）
<400> 1
tgcagtcgag cgatgaagct ccttcgggag tggattagcg gcggacgggt gagtaacacg 60
tgggtaacct gcctaataga gagggatagc cttccgaaag gaagattaat acctcataag 120
attgtagttt cgcatgaaac agcaataaaa ggagcaatcc gctatgagat ggacccgcgt 180
cgcattagct agttggtaag gtaaaggctt accaaggcga cgatgcgtag ccgacctgag 240
agggtgatcg gccacattgg gactgagaca cggcccagac tcctacggga ggcagcagtg 300
gggaatattg cacaatgggg gaaaccctga tgcagcaacg ccgcgtgagt gatgacggtc 360
ttcggattgt aaaactctgt ctttggggac gataatgacg gtacccaagg aggaagccac 420
ggctaactac gtgccagcag ccgcggtaat acgtaggtgg caagcgttgt ccggatttac 480
tgggcgtaaa gggagcgtag gtggatattt aagtgggatg tgaaatactc gggcttaacc 540
tgagtgctgc attccaaact ggatatctag agtgcaggag aggaaagtgg aattcctagt 600
gtagcggtga aatgcgtaga tattaggaag aacaccagtg gcgaaggcga ctttctggac 660
tgtaactgac actgaggctc gaaagcgtgg ggagcgaaca ggattagata ccctggtagt 720
ccacgccgta aacgatgaat actaggtgta ggggttgtca tgacctctgt gccgccgcta 780
acgcattaag tattccgcct ggggagtacg gtcgcaagat taaaactcaa aggaattgac 840
gggggcccgc acaagcagcg gagcatgtgg tttaattcga agcaacgcga agaaccttac 900
ctagacttga catctcctga attactctta atcgaggaag tcacttcggt gacaggaaga 960
caggtggtgc atggttgtcg tcagctcgtg tcgtgagatg ttgggttaag tcccgcaacg 1020
agcgcaaccc ttattgttag ttgctaccat taagttgagc actctagcga gactgcccgg 1080
gttaaccggg aggaaggtgg ggatgacgtc aaatcatcat gccccttatg tctagggcta 1140
cacacgtgct acaatggcaa gtacagagag atgctaaacc gtgaggtgga gccaaacttc 1200
aaaacttgtc tcagttcgga ttgtaggctg aaactcgcct acatgaagct ggagttgcta 1260
gtaatcgcga atcagaatgt cgcggtgaat acgttcccgg gccttgtaca caccgcccgt 1320
cacaccatga gagttggcaa tacccaaagt tcgtgagcta acccgtaagg gaggcagcga 1380
cct 1383
Claims (1)
1. a kind of method handling oil extraction waste water using YB-6 bacterial strain anaerobic degradations, it is characterised in that the specific steps are：
(1) YB-6 bacterial strains are enriched in reinforced clostridial medium, are placed in anaerobic culture box and are cultivated 7 days at 37 DEG C, be made
It is enriched with bacterium solution；
(2) step (1) enrichment bacterium solution obtained is inoculated into oil extraction waste water, the volume ratio for being enriched with bacterium solution and oil extraction waste water is 1:
150, then the constant-temperatureanaerobic anaerobic culture 96h at 37 DEG C, that is, complete the anaerobic degradation processing to oil extraction waste water；
The composition of the reinforced clostridial medium is as follows：9 ~ 11g/L of extracted beef powder, 9 ~ 11g/L of peptone, yeast extract powder 2 ~
4g/ L, 0.5 ~ 1.5g/L of soluble starch, 4 ~ 6g/L of glucose, 0.5 ~ 1g/L of cysteine hydrochloride, 4 ~ 6g/L of sodium chloride, acetic acid
2 ~ 4g/L of sodium；The pH value 6.8 ± 0.2 of the reinforced clostridial medium；
The YB-6 bacterial strains are now preserved in China General Microbiological culture presevation administrative center, preservation date：03 month 2016 15
Day, deposit number：CGMCC No.12214, Classification And Nomenclature：Clostridium Clostridium sp.；Its thalli morphology and molecular biosciences
Learn feature：Thalline is rod-shaped, 0.4 ~ 0.6 2.5 ~ 3.2 μm of μ m, single or arrangement in pairs, and raw in brood cell, column, cyst expands, and removes from office
Lan Shi is positive.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101054567A (en) * | 2007-04-06 | 2007-10-17 | 哈尔滨工业大学 | Clostridium Daqing clostridium and application thereof |
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2016
- 2016-07-19 CN CN201610566032.7A patent/CN106635864B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101054567A (en) * | 2007-04-06 | 2007-10-17 | 哈尔滨工业大学 | Clostridium Daqing clostridium and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| Culture-Independent Analysis of Bacterial Community Composition during Bioremediation of Crude Oil-Polluted Soil;Chioma Blaise Chikere;《British Microbiology Research Journal》;20121231;第2卷(第3期);第187-211页 * |
| 克鲁氏梭状芽孢杆菌的研究进展;丁雪松等;《食品工业科技》;20121231;第33卷(第15期);摘要及第403页左栏第2.3.1 * |
| 采油废水处理系统活性污泥中可培养菌群多样性研究;史利荣等;《环境科学与技术》;20140630;第37卷(第6期);第38-43页 * |
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