CN107058108A - A kind of method using improved folded ware flat band method screening separation sulfate reducing bacteria - Google Patents
A kind of method using improved folded ware flat band method screening separation sulfate reducing bacteria Download PDFInfo
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Abstract
The present invention relates to a kind of method using improved folded ware flat band method screening separation sulfate reducing bacteria, this method carries out switching screening by sulfate reduction bacteria liquid enriched medium to the microorganism in soil first, then using the dilution spread repeat dish sandwish method and screening and culturing medium culture bacterial strain after improving, 3rd step isolates and purifies previous step obtained strains using commonly folded ware double-layer agar technique, and culture is finally preserved in Storaged media.The method overcome prior art presence bacterium solution can not equably coating penetrating to be unfavorable for its growth, bacterium colony on first layer flat board, after bacterium solution dilution easily overlapping or even the deficiencies such as varied bacteria growing occur up and down, have the advantages that with material is easy, screening efficiency is high, the screening cycle is short, gained strain high conversion rate.
Description
Technical field
The invention belongs to technical field of environmental microorganism, and in particular to one kind utilizes improved folded ware flat band method screening separation
The method of sulfate reducing bacteria.
Background technology
At present, with high-tech biotechnologies such as bioengineering, plant and the soil that can be used in fixing heavy metal are cultivated
Microorganism super engineering bacteria, and apply to remediation contaminated soil be current environmental science, it is Environmental Engineering, plant physiology, micro-
One of most active Disciplinary Frontiers in the research such as biology.Microorganism can by redox, methylate and demethylation act on
Heavy metal is converted, noxious material is changed into the not available form of animals and plants.
Sulfate reducing bacteria (Sulfate-Reducing Bacterium, abbreviation SRB are similarly hereinafter) is can be sulphur in nature
The oxysulfides such as hydrochlorate, sulphite, thiosulfate and elemental sulphur reduction formation sulphion, due to heavy metal sulfide
Solubility product is universal smaller, and sulphion can generate biological unavailable form with micro heavy ions binding immediately, therefore
Sulfate reducing bacteria may be used as handling the heavy metal ion in fixed environment.SRB bacterium industrial pipeline burn into by heavy metal
Key player is all play in the waste water of pollution and the environment remediation of acidic mine waste water.
SRB is strict anaerobic bacteria, but going deep into research, existing result of study shows SRB not on stricti jurise
Absolute anaerobism but amphimicrobian, but SRB, to oxygen or extremely sensitive, therefore is cultivated with separating pass it in general
Key will use strict anaerobic technology.The available cultural methods of SRB have a lot, such as repeat dish sandwish method, dilution shake tube method,
Hungate rolling tube techniques, can also utilize anaerobism bag, anaerobic jar, anaerobism glove box etc..The weak point that tube method is shaken in dilution is to see
Examine relatively difficult with picking colony, Hungate rolls tube method and utilizes the method for anaerobism equipment then by technical method and equipment
The restriction of material cost, and repeat dish sandwish method be substantially by bacterium be clipped in above and below between two layers of culture medium, make its cause one it is relative
The environment of anaerobic, so that SRB can grow in crack.The advantage of repeat dish sandwish method is that culture is grown on battalion using coating
Support in agar interlayer, taking can easily accomplish that fixed point takes bacterium during bacterium colony, while this method need not create an anaerobic in addition
Environment, it is time saving, laborsaving, possess all aerobic, anaerobism separation method advantages while easily realization.
But traditional repeat dish sandwish method separation bacterium typically can not equably coating penetrating is on first layer flat board, and bacterium solution
Through dilution rear oxidation reduction potential can be raised with the increase of extension rate, for sulfate reducing bacteria growth very not
Profit.In addition dilution can be with diffusing into the second layer, so as to occur above and below bacterium colony during the pouring into of second layer agar layer
Phenomenon that is overlapping or even that the second layer surface of varied bacteria growing occur, is difficult to obtain single bacterium colony, and one is brought to separating and screening bacterium colony
Fixed difficulty.For the advantage of inheriting tradition Anaerobic culturel partition method, its shortcoming is overcome, the present invention is actually needed pair according to scientific research
The dilution spread of sulfate reducing bacteria (SRB)-repeat dish sandwish method isolated culture method is improved.
The content of the invention
The above mentioned problem that the present invention exists for existing sulfate reducing bacteria triage techniques is enriched with there is provided one kind from soil
And the sulfate reducing bacteria for being screened using the folded ware flat band method after improvement, separating, being purified into higher heavy metal ion tolerance
Method.This method operation is easier, improves the efficiency of screening separation, shortens screening separation cycle.It is above-mentioned to realize
Purpose, the technical solution adopted in the present invention is as follows：
A kind of method using improved folded ware flat band method screening separation sulfate reducing bacteria, comprises the following steps：(a) will
Pretreated soil, which is added in sulfate reduction bacteria liquid enriched medium, carries out Anaerobic culturel, and gained pregnant solution is seeded to
Anaerobic culturel is carried out in new sulfate reduction bacteria liquid enriched medium again, obtains being enriched with bacterium solution after switching culture repeatedly；
(b) enrichment bacterium solution is diluted to various concentrations, addition Cys, ascorbic acid and iron ammonium sulfate are standby thereto, will
Screening and culturing medium is poured into culture dish, the dilution of various concentrations is coated on flat board respectively after after its condensation, liquid to be diluted
Successively poured into twice after infiltration and the second layer, third layer are sequentially formed after screening and culturing medium, condensation, the closed rear constant temperature of culture dish is trained
Support；(c) screening and culturing medium after sterilizing is poured into culture dish condense it is standby, in picking step (b) second layer strain growth layer
Obvious black single bacterium is grown, putting down after line separation is poured into the flat lining out separation newly prepared, then by screening and culturing medium
It is repeated multiple times after sealing to be separately cultured on plate；(d) the isolated single bacterium colony that will rule is inoculated into Storaged media, constant temperature
Culture is produced.
According to such scheme, the composition of the sulfate reduction bacteria liquid enriched medium includes：0.4-0.6g/'s
K2HPO4, 1-2g/L MgSO4·7H2O, 0.4-0.6g/L Na2SO4, 0.1-0.2g/L CaCl2, 1-2g/L NaCl,
0.8-1.0g/L NH4Cl, 1.0-2.0g/L yeast extract, sterilize 20min after preparing at 121 DEG C, adjusts pH to 8.0-
8.5。
According to such scheme, the composition of the screening and culturing medium includes：0.4-0.6g/L K2HPO4, 0.1-0.2g/L's
CaCl2, 0.8-1.0g/L NH4Cl, 1-2g/L MgSO4·7H2O, 1-2g/L NaCl, 0.4-0.6g/L Na2SO4,
2.0-3.0g/L C3H5O3Na, 0.5-0.6g/L Cys, 0.5-0.6g/L ascorbic acid, 0.5-0.6g/L's
Fe(NH4)2(SO)2And 2wt% agar, first other raw materials are mixed and the 20min that sterilized at 121 DEG C during preparation, then dropped
Cys, ascorbic acid, Fe (NH are added after warm to 55 DEG C4)2(SO4)2, adjust pH to 8.0-8.5.
According to such scheme, the composition of the Storaged media includes：0.5-0.7g/L K2HPO4, 0.1-0.2g/L's
CaCl2, 1-1.2g/L NH4Cl, 1-2g/L MgSO4·7H2O, 2-3g/L NaCl, 0.4-0.6g/L Na2SO4,3.0-
4.0g/L C3H5O3Na, 0.5-0.6g/L Cys, 0.5-0.6g/L ascorbic acid are first former by other during preparation
Material mixing and the 20min that sterilized at 121 DEG C, be then cooled to after 55 DEG C add Cys, ascorbic acid, regulation pH to
8.0-8.5。
According to such scheme, screening pretreatment is carried out to pedotheque with screen cloth first in step (a), then according to 1g：
Soil is added in the sulfate reduction bacteria liquid enriched medium after sterilizing by 200-220mL ratio, in the covering of its surface
One layer of atoleine and inflated with nitrogen, are put into 30 DEG C of constant-temperature table and carry out Anaerobic culturel with 100-120r/min rotating speed.
According to such scheme, first enrichment time is 7d in step (a), afterwards with 5 ‰ inoculum concentration in new sulfate
Reduce switching culture 2-3 times, each duration 4d in bacteria liquid enriched medium.
According to such scheme, enrichment bacterium solution is diluted to 10 in step (b)-4、10-5、10-6Three concentration, then to dilution
0.5wt% Cys and 0.5wt% ascorbic acid and 0.5% iron ammonium sulfate are added in liquid.
According to such scheme, 55 DEG C of screening and culturing mediums of about 1/2 height are poured into step (b) into some culture dishes, it is cold
The dilution that 0.2-0.3mL various concentrations are drawn after solidifying is rapidly coating on each flat board, and liquid infiltration a period of time to be diluted is backward
The screening and culturing medium of the same race formation second layer strain growth layer of 1/3 height is poured into culture dish, then pours into culture medium of the same race and extremely will
Inexcessive form of overflowing forms third layer agar layer, and 30 DEG C of incubated 2-3d are placed in after covering the sealing of culture dish lid, until occurring
Obvious black colonies.
According to such scheme, line is separately cultured temperature for 30 DEG C in step (c), and line every time is separately cultured 3-4d, point
From 2-3 times.
According to such scheme, the isolated single bacterium colony of line is inoculated into by preservation culture using needle point method in step (d)
It is incubated in 30 DEG C in base, until occurring being placed in low temperature environment refrigeration after obvious filmanetous colony.
Compared with prior art, beneficial effects of the present invention are：
1) bacterium solution being coated on flat board can be dispersed in second layer Boping flaggy by the present invention well, be solved
The bacterium solution for being coated on first layer is easily mixed into second poured into afterwards by traditional folded ware flat band method when pouring into second layer agar layer
It is overlapping above and below bacterium colony so as to occur in layer agar layer, or even there is the phenomenon of varied bacteria growing in second layer agar layer surface；
2) the repeat dish sandwish method that the present invention is provided compares other improved method, and operation is easier, while two above and below because having
The layer seamless bubble-free of agar layer clamps intermediate strains grown layer, so that the growth to sulfate reducing bacteria is provided and preferably detested
Oxygen environment；
3) compared with current repeat dish sandwish method sulfate reducing bacteria method for screening and separating, compared with Traditional Method after present invention improvement
Apply to plant in same concentrations gradient and can filter out the sulfate reducing bacteria single bacterium colony of 50% quantity more, improve screening separation
Efficiency；
4) present invention because in dilution spread liquid addition 0.5wt% Cys and 0.5wt% ascorbic acid with
And 0.5wt% iron ammonium sulfate so that isolated sieve of the sulfate reducing bacteria compared with Traditional Method sulfate reducing bacteria of screening
Select and 2~3d is shortened in separation cycle.
Embodiment
To make those of ordinary skill in the art fully understand technical scheme and beneficial effect, below in conjunction with specific
Embodiment is further described.
Embodiment 1
The first step：The enrichment of sulfate reducing bacteria
It is first that the soil of sampling is standby with screen cloth removing bulky grain stone and plant roots and stems etc..Sulphur is prepared in beaker
Hydrochlorate reduces bacteria liquid enriched medium, and 200-220ml sulfate reduction bacteria liquid enrichment culture liquid is poured into conical flask,
In the 20min that sterilized at 121 DEG C in high-temperature sterilization pot.After it cools, weigh the soil after 1g is treated and be added to cooling
Into the conical flask of normal temperature, and in one layer of atoleine of conical flask nutrient solution Surface mulch, detest while being filled with nitrogen sealing holding
Oxygen environment, conical flask is put into 30 DEG C of constant-temperature tables and cultivated under 100~120r/min rotating speed.Enrichment time is for the first time
7d, until pregnant solution color to prepared Chinese ink color is to represent to be enriched with successfully, then with 5 ‰ inoculum concentration in new sulfate reduction bacterium solution
Switching culture 2-3 times, each duration 4d in body enriched medium.
Wherein, sulfate reduction bacteria liquid enriched medium includes following components：0.4-0.6g/ K2HPO4, 1-2g/L's
MgSO4·7H2O, 0.4-0.6g/L Na2SO4, 0.1-0.2g/L CaCl2, 1-2g/L NaCl, 0.8-1.0g/L's
NH4Cl, 1.0-2.0g/L yeast extract, sterilize 20min after preparing at 121 DEG C, adjusts pH to 8.0-8.5.
Second step：The dilution spread of bacterial strain-repeat dish sandwish method screening
The bacterium solution that above-mentioned acclimating is 2~3 times irradiates more than 15min through ultraviolet, and liquid-transfering gun is used on aseptic operating platform
Bacterium solution is diluted to 10-4、10-5、10-6Three concentration gradients, and into dilution add 0.5wt% Cys,
0.5wt% ascorbic acid and 0.5wt% iron ammonium sulfate.Then 55 DEG C will be maintained at after screening and culturing medium high-temperature sterilization
Left and right, about 1/2 height is poured into the ware bottom of some culture dishes, treats that its condensation is standby.Draw 0.2~0.3mL 10-4、10-5、
10-6The dilution of concentration gradient, respectively on rapidly coating some flat boards after condensation.Liquid to be diluted permeates about on flat board
After 10min, screening and culturing medium of the same race is poured slowly into the centre position at culture dish bottom to 1/3 height, bacterium solution is fixed therein
The second layer strain growth layer of formation, then screening and culturing medium of the same race is poured into the not excessive form formation third layer agar that will overflow
Layer.Cover culture dish lid rapidly to try not to leave bubble, remove agar unnecessary between two layers of side wall inside and outside culture dish, in latasuture
The paraffin melted in right amount is poured into, culture dish side parietal suture gap is sealed by paraffin, tries not to leave bubble.Finally culture dish is put
2-3d is cultivated in the insulating box for entering 30 DEG C, 10-520-30 black round point shape single bacterium is occurred in that in culture dish under concentration gradient
Fall, 10-62-4 black round point shape single bacterium colony is occurred in that in culture dish under concentration gradient.
Wherein, screening and culturing medium composition is：K2HPO40.4-0.6g/L, CaCl20.1-0.2g/L, NH4Cl 0.8-
1.0g/L, MgSO4·7H2O 1-2g/L, NaCl 1-2g/L, Na2SO40.4-0.6g/L, C3H5O3Na 2.0-3.0g/L, L-
Cysteine 0.5-0.6g/L, ascorbic acid 0.5-0.6g/L, Fe (NH4)2(SO)20.5-0.6g/L and 2wt% agar.
First other raw materials are mixed and the 20min that sterilized at 121 DEG C during preparation, is then cooled to after 55 DEG C and is added Cys, resists
Bad hematic acid, Fe (NH4)2(SO4)2, adjust pH to 8.0-8.5.
3rd step：Bacterial strain is isolated and purified
On aseptic operating platform, 55 DEG C or so will be maintained at after screening and culturing medium high-temperature sterilization, to some culture dish ware bottoms
In pour into about 1/2 height.After after its condensation, the culture medium of plate screening culture is sealed off, the fine jade that different cool times are solidified
Fat flat layer is carefully pried open, the obvious black single bacterium colony of picking and the laterally line separation in the flat board newly prepared.Again by high temperature
It is maintained at 55 DEG C or so of screening and culturing medium after sterilizing to pour on the flat board after line separation, until the inexcessive state that will overflow is rapid
Culture dish is covered, tries not to leave bubble, removes agar unnecessary between two layers of side wall inside and outside culture dish, the gap of culture dish is used
Rubber belt sealing.Culture dish is put into 30 DEG C of insulating box and cultivated, until there is obvious black origin shape single bacterium colony.Once draw
Line is separately cultured 3-4d, separates 2-3 times.
4th step：The preservation of bacterial strain
Storaged media and 10mL small test tubes are placed in 121 DEG C of sterilizing 20min, by Storaged media on aseptic operating platform
Pour into 10mL small test tubes, leave and take 1-2mm gap.After after its cooled and solidified with needle point method will rule separation after black list
It is pierced into after bacterium colony picking in the solid conservation culture medium after solidification, small test tube is put into 30 DEG C of constant temperature oven and cultivated, until
Occur being placed in deepfreeze after obvious filmanetous colony, storage life is up to more than half a year.
Wherein, Storaged media composition is：K2HPO40.5-0.7g/L, CaCl20.1-0.2g/L, NH4Cl1-1.2g/L,
MgSO4·7H2O 1-2g/L, NaCl2-3g/L, Na2SO40.4-0.6g/L, C3H5O3Na3.0-4.0g/L, Cys
0.5-0.6g/L, ascorbic acid 0.5-0.6g/L.First other raw materials are mixed and the 20min that sterilized at 121 DEG C during preparation, then
It is cooled to after 55 DEG C and adds Cys, ascorbic acid, adjusts pH to 8.0-8.5.
Screening gained sulfate reducing bacteria sulphate reducing ability is determined
Picking sulfate reducing bacteria is seeded in fluid nutrient medium from Storaged media, and bacterium is activated under anaerobic
Strain.In activating more than 4d under 100-120r/min rotating speed in 30 DEG C of constant-temperature tables, its OD is determined600After more than 0.5, press
Inoculum concentration according to 5 ‰ is forwarded in new fluid nutrient medium, and sub-methyl blue spectrum analysis is used after cultivating under anaerobic
Determine the S in bacterium solution2-Content, calculating obtains it by SO4 2-It is converted into S2-The conversion ratio of ion is 70%~80%.Wherein, detest
Oxygen condition is：Atoleine front cover is used, and is filled with nitrogen and keeps anaerobic state；Used fluid nutrient medium is constituted：K2HPO4
0.4-0.6g/L, NH4Cl0.8-1g/L, MgSO4·7H2O 1-2g/L, NaCl2-3g/L, Na2SO40.4-0.6g/L,
C3H5O3Na2.0-3.0g/L, L- half Guang ammonia 0.5-0.6g/L, ascorbic acid 0.5-0.6g/L.First other raw materials are mixed during preparation
Merge the 20min that sterilized at 121 DEG C, be then cooled to after 55 DEG C and add Cys, ascorbic acid, adjust pH to 8.0-
8.5。
Ware flat band method contrast test is folded by three stacking ware flat band methods after improvement with conventional double to find, in same concentrations
The sulfate reducing bacteria single bacterium colony quantity obtained under gradient has more 50% or so；And due to being added in dilution
The iron ammonium sulfate of 0.5wt% Cys, 0.5wt% ascorbic acid and 0.5wt%, is sulfate reducing bacteria
Growth creates good anaerobic environment, it is screened separation cycle and shortens 2-3d；Final determine shows simultaneously, screening gained
Strain is by SO4 2-It is converted into S2-The high conversion rate of ion is higher by 10%~20% up to 70%~80% than conventional screening methods.
Claims (10)
1. a kind of method using improved folded ware flat band method screening separation sulfate reducing bacteria, it is characterised in that including following
Step：(a) pretreated soil is added in sulfate reduction bacteria liquid enriched medium and carries out Anaerobic culturel, gained is rich
Liquid collecting is seeded in new sulfate reduction bacteria liquid enriched medium carries out Anaerobic culturel again, is obtained after switching culture repeatedly
It is enriched with bacterium solution；(b) enrichment bacterium solution is diluted to various concentrations, Cys, ascorbic acid and ferrous sulfate is added thereto
Ammonium is standby, and screening and culturing medium is poured into culture dish, and the dilution of various concentrations is coated on flat board respectively after after its condensation,
Successively poured into twice after liquid infiltration to be diluted and the second layer, third layer are sequentially formed after screening and culturing medium, condensation, culture dish is closed
It is incubated afterwards；(c) screening and culturing medium after sterilizing is poured into culture dish and condenses standby, picking step (b) second layer bacterial strain
Obvious black single bacterium is grown in grown layer, line point is poured into the flat lining out separation newly prepared, then by screening and culturing medium
It is repeated multiple times after sealing to be separately cultured on flat board from after；(d) the isolated single bacterium colony that will rule is inoculated into Storaged media
It is interior, it is incubated to produce.
2. a kind of method using improved folded ware flat band method screening separation sulfate reducing bacteria according to claim 1,
Characterized in that, the composition of the sulfate reduction bacteria liquid enriched medium includes：0.4-0.6g/ K2HPO4, 1-2g/L's
MgSO4·7H2O, 0.4-0.6g/L Na2SO4, 0.1-0.2g/L CaCl2, 1-2g/L NaCl, 0.8-1.0g/L's
NH4Cl, 1.0-2.0g/L yeast extract, sterilize 20min after preparing at 121 DEG C, adjusts pH to 8.0-8.5.
3. a kind of method using improved folded ware flat band method screening separation sulfate reducing bacteria according to claim 1,
Characterized in that, the composition of the screening and culturing medium includes：0.4-0.6g/L K2HPO4, 0.1-0.2g/L CaCl2, 0.8-
1.0g/L NH4Cl, 1-2g/L MgSO4·7H2O, 1-2g/L NaCl, 0.4-0.6g/L Na2SO4, 2.0-3.0g/L's
C3H5O3Na, 0.5-0.6g/L Cys, 0.5-0.6g/L ascorbic acid, 0.5-0.6g/L Fe (NH4)2(SO)2
And 2wt% agar, first other raw materials are mixed and the 20min that sterilized at 121 DEG C during preparation, is then cooled to after 55 DEG C and adds
Enter Cys, ascorbic acid, Fe (NH4)2(SO4)2, adjust pH to 8.0-8.5.
4. a kind of method using improved folded ware flat band method screening separation sulfate reducing bacteria according to claim 1,
Characterized in that, the composition of the Storaged media includes：0.5-0.7g/L K2HPO4, 0.1-0.2g/L CaCl2, 1-
1.2g/L NH4Cl, 1-2g/L MgSO4·7H2O, 2-3g/L NaCl, 0.4-0.6g/L Na2SO4, 3.0-4.0g/L's
C3H5O3Na, 0.5-0.6g/L Cys, 0.5-0.6g/L ascorbic acid first mix other raw materials simultaneously during preparation
Sterilize 20min at 121 DEG C, is then cooled to after 55 DEG C and adds Cys, ascorbic acid, adjusts pH to 8.0-8.5.
5. a kind of method using improved folded ware flat band method screening separation sulfate reducing bacteria according to claim 1,
It is characterized in that：Screening pretreatment is carried out to pedotheque with screen cloth first in step (a), then according to 1g：200-220mL's
Soil is added in the sulfate reduction bacteria liquid enriched medium after sterilizing by ratio, and one layer of atoleine is covered on its surface
And inflated with nitrogen, it is put into 30 DEG C of constant-temperature table and Anaerobic culturel is carried out with 100-120r/min rotating speed.
6. a kind of method using improved folded ware flat band method screening separation sulfate reducing bacteria according to claim 1,
It is characterized in that：First enrichment time is 7d in step (a), afterwards with 5 ‰ inoculum concentration in new sulfate reduction bacteria liquid
Switching culture 2-3 times, each duration 4d in enriched medium.
7. a kind of method using improved folded ware flat band method screening separation sulfate reducing bacteria according to claim 1,
It is characterized in that：Enrichment bacterium solution is diluted to 10 in step (b)-4、10-5、10-6Three concentration, are then added into dilution
The iron ammonium sulfate of 0.5wt% Cys and 0.5wt% ascorbic acid and 0.5wt%.
8. a kind of method using improved folded ware flat band method screening separation sulfate reducing bacteria according to claim 1,
It is characterized in that：55 DEG C of screening and culturing mediums of about 1/2 height are poured into step (b) into some culture dishes, are drawn after condensation
The dilution of 0.2-0.3mL various concentrations is rapidly coating on each flat board, in liquid infiltration a period of time backward culture dish to be diluted
Pouring into the screening and culturing medium of the same race formation second layer strain growth layer of 1/3 height, then pouring into culture medium of the same race will extremely overflow what is do not overflow
Form formation third layer agar layer, 30 DEG C of incubated 2-3d are placed in after covering the sealing of culture dish lid, until occurring obvious black
Color bacterium colony.
9. a kind of method using improved folded ware flat band method screening separation sulfate reducing bacteria according to claim 1,
It is characterized in that：Line is separately cultured temperature for 30 DEG C in step (c), and line every time is separately cultured 3-4d, separates 2-3 times.
10. a kind of method using improved folded ware flat band method screening separation sulfate reducing bacteria according to claim 1,
It is characterized in that：The isolated single bacterium colony of line is inoculated into Storaged media using needle point method in step (d), in 30 DEG C
It is incubated, until occurring being placed in low temperature environment refrigeration after obvious filmanetous colony.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110713899A (en) * | 2019-10-30 | 2020-01-21 | 南通大学 | Culture method for culturing desulfurization vibrio |
| CN111548940A (en) * | 2020-04-23 | 2020-08-18 | 华东理工大学 | Method for screening and producing biosurfactant facultative anaerobes |
| CN114058533A (en) * | 2021-09-30 | 2022-02-18 | 清华大学 | Screening method of sulfate reducing bacteria capable of degrading polycyclic aromatic hydrocarbons |
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| CN102260642A (en) * | 2011-07-20 | 2011-11-30 | 天津亿利科能源科技发展股份有限公司 | Method for separating and purifying sulfate reducing bacteria in oil field production water |
| CN104312962A (en) * | 2014-10-27 | 2015-01-28 | 中国石油化工股份有限公司 | Separation and purification method for sulfate reducing bacteria in sewage of oil field |
| CN104630097A (en) * | 2014-12-22 | 2015-05-20 | 韶关市桃林绿化科技有限公司 | Acidophilus sulfate reducing bacterium strain and application thereof |
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| CN110713899B (en) * | 2019-10-30 | 2023-04-28 | 南通大学 | Culture method for culturing vibrio desulphurisation |
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| CN111548940B (en) * | 2020-04-23 | 2023-02-10 | 华东理工大学 | Method for screening and producing biosurfactant facultative anaerobes |
| CN114058533A (en) * | 2021-09-30 | 2022-02-18 | 清华大学 | Screening method of sulfate reducing bacteria capable of degrading polycyclic aromatic hydrocarbons |
| CN114058533B (en) * | 2021-09-30 | 2023-10-03 | 清华大学 | Screening method of sulfate reducing bacteria capable of degrading polycyclic aromatic hydrocarbon |
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