CN106318871A - Strain screening method with efficient desulfurization and denitrification activities simultaneously - Google Patents

Strain screening method with efficient desulfurization and denitrification activities simultaneously Download PDF

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CN106318871A
CN106318871A CN201610911519.4A CN201610911519A CN106318871A CN 106318871 A CN106318871 A CN 106318871A CN 201610911519 A CN201610911519 A CN 201610911519A CN 106318871 A CN106318871 A CN 106318871A
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bacterial strain
culture medium
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screening method
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CN106318871B (en
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王晓慧
郭爱洪
陈健飞
吕德东
刘璐
席琦
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China Petroleum and Chemical Corp
Technology Inspection Center of Sinopec Shengli Oilfield Co
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China Petroleum and Chemical Corp
Technology Inspection Center of Sinopec Shengli Oilfield Co
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Abstract

The invention provides a strain screening method with efficient desulfurization and denitrification activities simultaneously and belongs to the technical field of biology. The method comprises the following steps: primary screening in the first step, purification and separation in the second step, biochemical culture in the third step and multiplication culture in the fourth step. According to the screening method provided by the invention, by setting screening and culturing conditions, strains with relatively high desulfurization and denitrification activities can be obtained. For strains obtained by adopting the screening method, the sulfide degradation efficiency is higher than 99%, and the ammonia nitrogen degradation efficiency is higher than 70%. The strain can save water treatment facility investment, ensure the sewage treatment effect, reduce investment for chemical treatment and energy consumption, and lower the sewage treatment cost, which is at least lowered by 0.20 yuan/cubic meter, has no secondary pollution, and can finally eliminate the influence to surrounding water environment and atmospheric environment; furthermore, the strain can be applied to treatment of petrochemical wastewater and domestic sewage, and has good environment and social benefits.

Description

A kind of bacterial strain screening method with high-efficiency desulfurization denitrification activity simultaneously
Technical field
The present invention relates to biological technical field, particularly relate to a kind of bacterial strain screening with high-efficiency desulfurization denitrification activity simultaneously Method.
Background technology
Current oil field produced water handling reclamation main object is petroleum-type, ammonia nitrogen, sulfide: sulfide is to cause oil field to note One of principal element of corrosion phenomenon serious in water system and industrial circulating cooling water；Ammonia nitrogen causes body eutrophication One of material.
Containing a large amount of H in oil plant treatment of wastewater from stripping2S and NH3, unprocessed in line enter to purify between waterwheel, certainly will be raw to routine Change system impacts, and causes always discharging the defective discharge of water；The water pot volatilization gas stench of acidic water stripping device is serious, and Exacerbate tank deck and the corrosion of neighbouring equipment.
Biological desulfurization process is because clearance is high, without adding chemical agent, can generate elemental sulfur recovery mineral nitrogen resource, operation The advantages such as expense formation secondary pollution low, less and of increasing concern, it has also become remove sulphide removal and odor prevention both at home and abroad Main stream approach in application；Bio-denitrification technology is owing to having high treating effect, and processing procedure is reliable and stable, and processing cost is low, The advantages such as convenient operation and management and be used widely, the removal for Water provides effective means.
The bacterial strain that some current screening techniques obtain can realize the desulfurization to sewage disposal or denitrogenation, and for same The waste water of Shi Hanyou sulfur and nitrogen carries out desulfurization and denitrogenation, and at present, the application carried out a biological disposal upon in water processes is concentrated mainly on: 1) good Oxygen desulfurization；2) anaerobism desulfurization；3) the anti-nitre of anaerobism.On the one hand add operating cost, on the other hand add equipment and raw material Put into, and synchronized desulfuring denitrogenation becomes a kind of brand-new research direction, also not about utilizing biological treatment simultaneous desulfurization and denitrification, And desulfurization product is controlled the relevant report in the elemental sulfur stage.
Summary of the invention
It is an object of the invention to provide a kind of bacterial strain screening method with high-efficiency desulfurization denitrification activity simultaneously.
For achieving the above object, the present invention is realized by following measure: one has high-efficiency desulfurization simultaneously and takes off Nitrogen activity bacterial strain screening method, comprise the following steps: first step primary dcreening operation, second step purifies and separates, the 3rd step biochemical culture and 4th step enrichment culture；
The program of first step primary dcreening operation is:
Take 20-30g activated sludge and put in the culture dish filling primary dcreening operation culture medium, culture dish is put into 25-30 DEG C of constant-temperature table Middle cultivation, the rotating speed keeping constant-temperature table is that 180-185 turns/min, and incubation time is 4-5 days；
Primary dcreening operation culture medium is by 13-15g Na2S2O3、7-9g KNO3、5-6g (NH4)2HPO4、20-1.5g MgCl2、5-6g NaHCO3, 1-1.5g cysteine, 25-30ml vitamin liquid and 25-30ml microelement nutritious liquid mixing, regulation pH be 7.0, and through 120-121 DEG C, obtain after the sterilization treatment of 18-20min；
The program of second step purifies and separates is:
The strain in culture dish after being cultivated by first step shaking table is seeded to solid medium, by the solid culture after inoculation strain Base is placed on the biochemical cultivation case of 25-30 DEG C and cultivates 2-3 days；
Solid medium is by 12-15g Na2S2O3、5-7g KNO3、7-9g (NH4)2HPO4、1-1.5g MgCl2、6-7g NaHCO3, 1.3-1.5g cysteine, 10-12g agar, 0.1-0.15g activated carbon, 1.8-2g mannose, 28-30ml vitamin Nutritional solution and 28-30ml microelement nutritious liquid mix；
The program of the 3rd step biochemical culture is:
Put respectively to primary dcreening operation culture medium, at 25-from single bacterium colony of the line isolated of picking through the culture medium of second step 30 DEG C, 180-185 turn/shaking table of min cultivates 8-10 days, takes 2ml bacterium solution to being placed with 100-150ml anaerobic culture medium after cultivation In anaerobism bottle, then anaerobism bottle is put into 25-30 DEG C of biochemical cultivation case 25-30 days；Again anaerobism bottle is put to 30 DEG C of Anaerobic culturel In tank, cultivate 25 days, during cultivating, fill the N of 5min2Or persistently rush N2,Obtain bacterial strain；
Anaerobic culture medium is by 0.02-0.025g MnSO4、10-12g Na2S2O3、1-1.2g Na2HPO4、1.5-1.8g KH2HPO4、1-1.5g NaHCO3、0.4-0.5g MgSO4•7H2O、0.5-0.6g NH4Cl、0.05-0.1g CaCl2、0.02- 0.05g FeCl3With 5-5.5g KNO3Boil after mixing and dry in the air cool, add and adjust after 0.5 ‰ cysteine pH to neutral, then add 1- 1.2ml indicator, fills N2Bottling, through 120-121 DEG C, the sterilizing of 20-25min obtains；
The program of the 4th step enrichment culture is:
Each bacterial strain is inoculated into respectively equipped with carrying out enrichment culture in the denitrification and desulfurization culture medium of 130-150ml, puts into 35-37 DEG C Shaken cultivation case is cultivated 7 days, obtains having higher desulfurization, the bacterial strain of fall nitrogen activity；
Denitrification and desulfurization culture medium is by 10-12g peptone, 3-3.5g Carnis Bovis seu Bubali cream, 5-6g NaCl, 1-1.5g KNO3, 1-2mg hexamethylene Six alcohol, 3-6mg6-benayl aminopurine, 0.5-0.8g cysteine and the mixing of 20-25g agar, and adjust pH to obtain to 7.2.
Wherein, in described vitamin liquid thiamine be 0.12-0.15ug/L, pyridoxol be 0.13-0.15ug/L, general Acid calcium is 0.15-0.2ug/L and nicotinic acid is 0.11-0.15ug/L.
Wherein, the composition of described microelement nutritious liquid be potassium iodide 0.5-0.65mg/L, zinc sulfate 5-7mg/L, chlorination Cobalt 0.02-0.05mg/L, copper sulfate 0.02-0.05mg/L, biotin 0.3-0.5mg/L, methionine 0.2-0.5mg/L, Soviet Union's ammonia The mixed solution of acid 0.28-0.3mg/L.
Wherein, described indicator is methylene blue Anaerobic indicator.
Described activated sludge is oil field produced water treatment plant biochemical sludge, ash field biochemical sludge or sewage treatment plants Aeration tank.
The invention have the benefit that pick-ups and deliveries under the bacterial strain anaerobic condition that this screening technique obtains, have relatively High lasting sulfide degrading activity and ammonia nitrogen degradation activity, sulfide degradation rate reaches more than 99%, and ammonia nitrogen degradation rate reaches More than 70%；The bacterial strain that this screening technique obtains is applied in sewage disposal without reactor is carried out aeration, it is possible to decrease run into This；Without additional Organic substance as electron acceptor, both reduced cost and turn avoid the load increasing reactor；This screening technique obtains To bacterial strain achieve sulfide and be converted into elemental sulfur and can carry out resource reclaim, prevent from causing secondary pollution；And this screening side The bacterial strain that method obtains, can apply to the process of sanitary sewage and petrifaction sewage, obtains good economic benefit, has the most wide Market.
Accompanying drawing explanation
Fig. 1 be the embodiment of the present invention 1 about ammonia nitrogen, sulfide time dependent degradation efficiency figure.
Fig. 2 be the embodiment of the present invention 2 about ammonia nitrogen, sulfide time dependent degradation efficiency figure.
Fig. 3 be the embodiment of the present invention 3 about ammonia nitrogen, sulfide time dependent degradation efficiency figure.
Detailed description of the invention
For the technical characterstic of this programme can be clearly described, below by detailed description of the invention, this programme is illustrated.
Embodiment 1
The program of first step primary dcreening operation is:
The biochemical sludge taking 20g waste ash pond is put in the culture dish filling primary dcreening operation culture medium, and culture dish is put into 30 DEG C of constant temperature Cultivating in shaking table, the rotating speed keeping constant-temperature table is 185 turns/min, and incubation time is 5 days；
Primary dcreening operation culture medium is by 15g Na2S2O3、9g KNO3、6g (NH4)2HPO4、1.5g MgCl2、6g NaHCO3, 1.5g half Guang Propylhomoserin, 30ml vitamin liquid and the mixing of 30ml microelement nutritious liquid, regulation pH is 7.0, and through 121 DEG C, 20min Obtain after sterilization treatment；
The program of second step purifies and separates is:
The strain in culture dish after being cultivated by first step shaking table is seeded to solid medium, by the solid culture after inoculation strain Base is placed on the biochemical cultivation case of 30 DEG C and cultivates 3 days；
Solid medium is by 15g Na2S2O3、9g KNO3、6g (NH4)2HPO4、1.5g MgCl2、6g NaHCO3, 1.5g half Guang Propylhomoserin, 12g agar, 0.1g activated carbon, 2g mannose, 30ml vitamin liquid and 30ml microelement nutritious liquid mixing and Become；
The program of the 3rd step biochemical culture is:
Put respectively to primary dcreening operation culture medium, 30 from single bacterium colony of the line isolated of picking through the culture medium of second step DEG C, the shaking table of 185 turns/min cultivate 10 days, take 2ml bacterium solution after cultivation in the anaerobism bottle being placed with 150ml anaerobic culture medium, so After anaerobism bottle is put into 30 DEG C of biochemical cultivation cases 30 days；Anaerobism bottle is put to 30 DEG C of anaerobic jars again, cultivate 25 days, in training Persistently rush N2 during Yanging, obtain bacterial strain；
Anaerobic culture medium is by 0.02g MnSO4、10g Na2S2O3、1.2g Na2HPO4、1.8g KH2HPO4、1g NaHCO3、0.4g MgSO4•7H2O、0.5g NH4Cl、0.05g CaCl2、0.02g FeCl3With 5g KNO3Boil after mixing and dry in the air cool, add 0.5 ‰ half Adjust pH to neutral after cystine, then add 1ml methylene blue Anaerobic indicator, fill N2Bottling, through 121 DEG C, the sterilizing of 25min obtains Arrive；
The program of the 4th step enrichment culture is:
Each bacterial strain is inoculated into respectively equipped with the denitrification and desulfurization culture medium of 150 ml carries out enrichment culture, puts into 37 DEG C of vibration trainings Support in case and cultivate 7 days, obtain having higher desulfurization, fall nitrogen activity bacterial strain 1.；
Denitrification and desulfurization culture medium is by 10g peptone, 3g Carnis Bovis seu Bubali cream, 5g NaCl, 1g KNO3, 2mg cyclohexanhexanol, 4mg 6-benzyl ammonia Base purine, 0.5g cysteine and the mixing of 20g agar, and adjust pH to obtain to 7.2.
Wherein, in described vitamin liquid thiamine be 0.12ug/L, pyridoxol be 0.13ug/L, calcium pantothenate be 0.15ug/L and nicotinic acid are 0.11ug/L.
Wherein, the composition of described microelement nutritious liquid be potassium iodide 0.65mg/L, zinc sulfate 5mg/L, cobaltous chloride 0.02mg/L, copper sulfate 0.02mg/L, biotin 0.3mg/L, methionine 0.2mg/L, the mixed solution of threonine 0.28mg/L.
Embodiment 2
The program of first step primary dcreening operation is:
Qu20gSha Ying sewage treatment plant biochemical sludge is put in the culture dish filling primary dcreening operation culture medium, is put into by culture dish Cultivating in 30 DEG C of constant-temperature tables, the rotating speed keeping constant-temperature table is 185 turns/min, and incubation time is 5 days；
Primary dcreening operation culture medium is by 15g Na2S2O3、9g KNO3、6g (NH4)2HPO4、1.5g MgCl2、6g NaHCO3, 1.5g half Guang Propylhomoserin, 30ml vitamin liquid and the mixing of 30ml microelement nutritious liquid, regulation pH is 7.0, and through 121 DEG C, 20min Obtain after sterilization treatment；
The program of second step purifies and separates is:
The strain in culture dish after being cultivated by first step shaking table is seeded to solid medium, by the solid culture after inoculation strain Base is placed on the biochemical cultivation case of 30 DEG C and cultivates 3 days；
Solid medium is by 15g Na2S2O3、9g KNO3、6g (NH4)2HPO4、1.5g MgCl2、6g NaHCO3, 1.5g half Guang Propylhomoserin, 12g agar, 0.1g activated carbon, 2g mannose, 30ml vitamin liquid and 30ml microelement nutritious liquid mixing and Become；
The program of the 3rd step biochemical culture is:
Put respectively to primary dcreening operation culture medium, 30 from single bacterium colony of the line isolated of picking through the culture medium of second step DEG C, the shaking table of 185 turns/min cultivate 10 days, take 2ml bacterium solution after cultivation in the anaerobism bottle being placed with 150ml anaerobic culture medium, so After anaerobism bottle is put into 30 DEG C of biochemical cultivation cases 30 days；Anaerobism bottle is put to 30 DEG C of anaerobic jars again, cultivate 25 days, in training Fill the N2 of 5min during Yanging, obtain bacterial strain；
Anaerobic culture medium is by 0.02g MnSO4、10g Na2S2O3、1.2g Na2HPO4、1.8g KH2HPO4、1g NaHCO3、0.4g MgSO4•7H2O、0.5g NH4Cl、0.05g CaCl2、0.02g FeCl3With 5g KNO3Boil after mixing and dry in the air cool, add 0.5 ‰ half Adjust pH to neutral after cystine, then add 1ml methylene blue Anaerobic indicator, fill N2Bottling, through 121 DEG C, the sterilizing of 25min obtains Arrive；
The program of the 4th step enrichment culture is:
Each bacterial strain is inoculated into respectively equipped with the denitrification and desulfurization culture medium of 150 ml carries out enrichment culture, puts into 37 DEG C of vibration trainings Support in case and cultivate 7 days, obtain having higher desulfurization, fall nitrogen activity bacterial strain 2.；
Denitrification and desulfurization culture medium is by 10g peptone, 3g Carnis Bovis seu Bubali cream, 5g NaCl, 1g KNO3, 2mg cyclohexanhexanol, 4mg 6-benzyl ammonia Base purine, 0.5g cysteine and the mixing of 20g agar, and adjust pH to obtain to 7.2.
Wherein, in described vitamin liquid thiamine be 0.12ug/L, pyridoxol be 0.13ug/L, calcium pantothenate be 0.15ug/L and nicotinic acid are 0.11ug/L.
Wherein, the composition of described microelement nutritious liquid be potassium iodide 0.65mg/L, zinc sulfate 5mg/L, cobaltous chloride 0.02mg/L, copper sulfate 0.02mg/L, biotin 0.3mg/L, methionine 0.2mg/L, the mixed solution of threonine 0.28mg/L.
Embodiment 3
The program of first step primary dcreening operation is:
The activated sludge taking 30g stake western sewage disposal plant aeration tank is put in the culture dish filling primary dcreening operation culture medium, by culture dish Putting in 30 DEG C of constant-temperature tables and cultivate, the rotating speed keeping constant-temperature table is 185 turns/min, and incubation time is 5 days；
Primary dcreening operation culture medium is by 15g Na2S2O3、9g KNO3、6g (NH4)2HPO4、1.5g MgCl2、6g NaHCO3, 1.5g half Guang Propylhomoserin, 30ml vitamin liquid and the mixing of 30ml microelement nutritious liquid, regulation pH is 7.0, and through 121 DEG C, 20min Obtain after sterilization treatment；
The program of second step purifies and separates is:
The strain in culture dish after being cultivated by first step shaking table is seeded to solid medium, by the solid culture after inoculation strain Base is placed on the biochemical cultivation case of 30 DEG C and cultivates 3 days；
Solid medium is by 15g Na2S2O3、9g KNO3、6g (NH4)2HPO4、1.5g MgCl2、6g NaHCO3, 1.5g half Guang Propylhomoserin, 12g agar, 0.1g activated carbon, 2g mannose, 30ml vitamin liquid and 30ml microelement nutritious liquid mixing and Become；
The program of the 3rd step biochemical culture is:
Put respectively to primary dcreening operation culture medium, 30 from single bacterium colony of the line isolated of picking through the culture medium of second step DEG C, the shaking table of 185 turns/min cultivate 10 days, take 2ml bacterium solution after cultivation in the anaerobism bottle being placed with 150ml anaerobic culture medium, so After anaerobism bottle is put into 30 DEG C of biochemical cultivation cases 30 days；Anaerobism bottle is put to 30 DEG C of anaerobic jars again, cultivate 25 days, in training Fill the N2 of 5min during Yanging or persistently rush N2, obtaining bacterial strain；
Anaerobic culture medium is by 0.02g MnSO4、10g Na2S2O3、1.2g Na2HPO4、1.8g KH2HPO4、1g NaHCO3、0.4g MgSO4•7H2O、0.5g NH4Cl、0.05g CaCl2、0.02g FeCl3With 5g KNO3Boil after mixing and dry in the air cool, add 0.5 ‰ half Adjust pH to neutral after cystine, then add 1ml methylene blue Anaerobic indicator, fill N2Bottling, through 121 DEG C, the sterilizing of 25min obtains Arrive；
The program of the 4th step enrichment culture is:
Each bacterial strain is inoculated into respectively equipped with the denitrification and desulfurization culture medium of 150 ml carries out enrichment culture, puts into 37 DEG C of vibration trainings Support in case and cultivate 7 days, obtain having higher desulfurization, fall nitrogen activity bacterial strain 3.；
Denitrification and desulfurization culture medium is by 10g peptone, 3g Carnis Bovis seu Bubali cream, 5g NaCl, 1g KNO3, 2mg cyclohexanhexanol, 4mg 6-benzyl ammonia Base purine, 0.5g cysteine and the mixing of 20g agar, and adjust pH to obtain to 7.2.
Wherein, in described vitamin liquid thiamine be 0.12ug/L, pyridoxol be 0.13ug/L, calcium pantothenate be 0.15ug/L and nicotinic acid are 0.11ug/L.
Wherein, the composition of described microelement nutritious liquid be potassium iodide 0.65mg/L, zinc sulfate 5mg/L, cobaltous chloride 0.02mg/L, copper sulfate 0.02mg/L, biotin 0.3mg/L, methionine 0.2mg/L, the mixed solution of threonine 0.28mg/L.
Degradation rate is tested
1. experimental subject
The bacterial strain that embodiments of the invention 1-3 obtains 1.-3..
2. experiment main agents
(1) ammonium sulfate [(NH4)2SO4], sodium nitrite (NaNO2), potassium nitrate (KNO3) and peptone be purchased from Beijing chemical industry respectively Factory, Beijing Yili Fine Chemicals Co., Ltd., Red Star chemical plant, Beijing and Beijing bispin microbiological culture media products factory.
(2) Griess reagent A liquid:
P-aminobenzene sulfonic acid 0.5 g；Spirit of vinegar (10%) 150ml
B liquid: alpha-naphthylamine 0.1 g, spirit of vinegar (10%) 150ml, H2O 20ml
(3) diphenylamines reagent:
Diphenylamines 0.5g；Concentrated sulphuric acid 100ml；Distilled water 20ml
(4) Gram stain:
1. ammonium oxalate crystal violet mixed liquor
Solution A: crystal violet 2.0g, ethanol (95%) 20ml
Second liquid: ammonium oxalate 0.8g, distilled water 80ml
First, second two liquid phase is mixed, filters after standing 48 hours and use.
2. Lu's grignard iodine liquid (iodine liquid)
Iodine 1.0g, potassium iodide 2.0g, distilled water 300ml, first dissolve potassium iodide with a small amount of distilled water 3-5ml, then put into Iodine tablet, adds water after iodine all dissolves and is settled to 300ml.
3. sarranine redyes liquid
Sarranine (Safranine O), 2.0g distilled water 100ml,
(5) C.I. 13020. reagent (M-R test reagent):
C.I. 13020. 0.1g, ethanol (95%) 300ml, H2O200ml
3. test key instrument equipment
Centrifuge (1-14, Germany Sigma), gel imaging system (GELDOC, U.S. Bio-Rad), optical microscope (BX51, Japan OLYMPUS), scanning electron microscope (JSM-6700, Japan JEOL), superclean bench (PCV, Japan HITACHI), ultraviolet-uisible spectrophotometer (Cary, VARIAN), autoclave (MLS-3750, Japan SANYO), incubator (MLR-350HT, Japan SANYO), Water Test Kits (HACH, U.S. DR2800)
4. experimental procedure
Fall nitrogen, desulphurizing activated detection
Take respectively 1.-3. in the bacterium solution 2ml to a of enrichment culture, tri-15ml denitrification and desulfurization test mediums of b, c, close plug 30 DEG C Lower cultivation two days.
Sampling from certain oil plant treatment of wastewater from stripping, recording the sulfide content in this oil plant treatment of wastewater from stripping is 45mg/L, ammonia Nitrogen content is 80mg/L, is respectively put into by waste water in three bottles, every bottle of 15ml.
Being respectively connected to the bacterium solution of 2ml a, b, c in three bottles, close plug room temperature (26 DEG C) is placed, every 24 h, centrifuging and taking Supernatant measures before and after cultivating ammonia nitrogen and sulfide content in waste water, and experiment carries out 4d, calculates each bacterial strain ammonia nitrogen and sulfide Removal efficiency.Ammonia nitrogen determination uses Nessler's reagent photometer；Sulfide measures and uses P-aminodimethylaniline photometry.
5. experimental result
Result of the test is shown in Table 1
By table 1 and accompanying drawing 1-3 it is known that the bacterial strain that the screening technique of the present invention obtains processes sewage, process the time shorter, Within 48 hours, just reaching reasonable nitrogen and desulfurization degradation effect, through processing, the concentration of sulfide and ammonia nitrogen compound is substantially reduced, Sulfide degradation rate reaches more than 99%, and ammonia nitrogen compound degradation rate reaches more than 70%, the bacterial strain that this screening technique of process obtains Processing, the sulfur nitrogen content of sewage reaches reduced levels.
The present invention can pass through or use prior art to realize without the technical characteristic described, and does not repeats them here, certainly, Described above is not limitation of the present invention, and the present invention is also not limited to the example above, the ordinary skill of the art Change that personnel are made in the essential scope of the present invention, retrofit, add or replace, also should belong to the protection model of the present invention Enclose.

Claims (5)

1. a bacterial strain screening method with high-efficiency desulfurization denitrification activity simultaneously, it is characterised in that comprise the following steps: first Step primary dcreening operation, second step purifies and separates, the 3rd step biochemical culture and the 4th step enrichment culture；
The program of first step primary dcreening operation is:
Take 20-30g activated sludge and put in the culture dish filling primary dcreening operation culture medium, culture dish is put into 25-30 DEG C of constant-temperature table Middle cultivation, the rotating speed keeping constant-temperature table is that 180-185 turns/min, and incubation time is 4-5 days；
Primary dcreening operation culture medium is by 13-15g Na2S2O3、7-9g KNO3、5-6g (NH4)2HPO4、20-1.5g MgCl2、5- 6gNaHCO3, 1-1.5g cysteine, 25-30ml vitamin liquid and 25-30ml microelement nutritious liquid mixing, regulate pH Be 7.0, and through 120-121 DEG C, obtain after the sterilization treatment of 18-20min；
The program of second step purifies and separates is:
The strain in culture dish after being cultivated by first step shaking table is seeded to solid medium, by the solid culture after inoculation strain Base is placed on the biochemical cultivation case of 25-30 DEG C and cultivates 2-3 days；
Solid medium is by 12-15g Na2S2O3、5-7g KNO3、7-9g (NH4)2HPO4、1-1.5g MgCl2、6-7g NaHCO3, 1.3-1.5g cysteine, 10-12g agar, 0.1-0.15g activated carbon, 1.8-2g mannose, 28-30ml vitamin Nutritional solution and 28-30ml microelement nutritious liquid mix；
The program of the 3rd step biochemical culture is:
Put respectively to primary dcreening operation culture medium, at 25-from single bacterium colony of the line isolated of picking through the culture medium of second step 30 DEG C, 180-185 turn/shaking table of min cultivates 8-10 days, takes 2ml bacterium solution to being placed with 100-150ml anaerobic culture medium after cultivation In anaerobism bottle, then anaerobism bottle is put into 25-30 DEG C of biochemical cultivation case 25-30 days；Again anaerobism bottle is put to 30 DEG C of Anaerobic culturel In tank, cultivate 25 days, during cultivating, fill the N of 5min2Or persistently rush N2,Obtain bacterial strain；
Anaerobic culture medium is by 0.02-0.025g MnSO4、10-12g Na2S2O3、1-1.2g Na2HPO4、1.5-1.8g KH2HPO4、1-1.5g NaHCO3、0.4-0.5g MgSO4•7H2O、0.5-0.6g NH4Cl、0.05-0.1g CaCl2、0.02- 0.05g FeCl3With 5-5.5g KNO3Boil after mixing and dry in the air cool, add and adjust after 0.5 ‰ cysteine pH to neutral, then add 1- 1.2ml indicator, fills N2Bottling, through 120-121 DEG C, the sterilizing of 20-25min obtains；
The program of the 4th step enrichment culture is:
Each bacterial strain is inoculated into respectively equipped with carrying out enrichment culture in the denitrification and desulfurization culture medium of 130-150 ml, puts into 35-37 DEG C shaken cultivation case is cultivated 7 days, the bacterial strain of desulfurization fall nitrogen activity while of obtaining having higher；
Denitrification and desulfurization culture medium is by 10-12g peptone, 3-3.5g Carnis Bovis seu Bubali cream, 5-6g NaCl, 1-1.5g KNO3, 1-2mg hexamethylene Six alcohol, 3-6mg 6-benzyl aminopurine, 0.5-0.8g cysteine and the mixing of 20-25g agar, and adjust pH to obtain to 7.2.
The bacterial strain screening method with high-efficiency desulfurization denitrification activity simultaneously the most according to claim 1, it is characterised in that institute State thiamine in vitamin liquid be 0.12-0.15ug/L, pyridoxol be 0.13-0.15ug/L, calcium pantothenate be 0.15- 0.2ug/L and nicotinic acid are 0.11-0.15ug/L.
The bacterial strain screening method with high-efficiency desulfurization denitrification activity simultaneously the most according to claim 1 and 2, its feature exists In, the composition of described microelement nutritious liquid is potassium iodide 0.5-0.65mg/L, zinc sulfate 5-7mg/L, cobaltous chloride 0.02- 0.05mg/L, copper sulfate 0.02-0.05mg/L, biotin 0.3-0.5mg/L, methionine 0.2-0.5mg/L, threonine 0.28- 0.3mg/L。
4., according to the bacterial strain screening method with high-efficiency desulfurization denitrification activity simultaneously described in any one of claim 1-3, it is special Levying and be, described indicator is methylene blue Anaerobic indicator.
5., according to the bacterial strain screening method with high-efficiency desulfurization denitrification activity simultaneously described in any one of claim 1-4, it is special Levying and be, described activated sludge is oil field produced water treatment plant biochemical sludge, ash field biochemical sludge or sewage treatment plants Aeration tank.
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