CN1786184A - Method of separating screening heterotrophic nitration bacteria - Google Patents
Method of separating screening heterotrophic nitration bacteria Download PDFInfo
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Abstract
The invention relates to a method of separating and scalping heterotrophy nitrobacteria. Its making steps are preparing beef extract-peptone liquid medium; heating sterilization after adding agar; condensing into flat solid medium; coating mixed microorganism flora to beef extract-peptone flat; setting in incubator at 30 centigrade degree to culture for 7-10 days; selecting mono bacteria to fall on the extract-peptone flat line to purify to from pure bacteria; inoculating the pure bacteria lawn to ammonia oxidation ability identification culture solution; dipping gelisi reagent into the culture solution to do nitration activity identification. The upper steps can separate heterotrophy nitrobacteria from the mixed microorganism series. So the invention is exact and reliable.
Description
Technical field
The present invention relates to a kind of method of separating screening heterotrophic nitration bacteria, be specifically related to a kind of from handle artificial simcity sanitary sewage system separation screening and identify the method for the heterotrophic organism with nitrification activity, belong to microbial technology field.
Background technology
In physical environment, nitrification mainly is to rely on the nitrated ability of microorganism to finish, and the nitrated notion of mentioning at present is meant that generally ammonia nitrogen is oxidized to nitrite nitrogen, further is oxidized to the process of nitrate nitrogen again.This process is finished by the different bacterium of two classes: a class is an ammonia oxidizing bacteria, and ammonia nitrogen is oxidized to nitrite nitrogen; Another kind of is nitrite-oxidizing bacteria, and nitrite nitrogen is oxidized to nitrate nitrogen.The main body that generally believes nitrification at present is the bacterial classification of chemosynthetic autotroph.
In recent years, Chinese scholars has been carried out a large amount of theories and experimental study to the problem and the phenomenon that disclose in the wastewater biological denitrificaion engineering practice, make every effort to further improve cognition to the nitrogen transformation process, and the new knowledge that some break through traditional theories has been proposed, wherein just comprise the proposition of heterotrophic nitrification notion.Think in the heterotrophic nitrification notion that ammonia nitrogen not only can finish traditional nitrifying process by autotrophic bacteria, some heterotrophic bacterium also can carry out the oxygenizement to ammonia nitrogen, and the substrate of heterotrophic nitrification effect is wider, both can be that inorganic nitrogen also can be an organic nitrogen, even can be nitrate nitrogen with the organonitrogen direct oxidation, be decomposed into ammonia nitrogen, reoxidize and be two steps of nitrite nitrogen and stride across organonitrogen.For the autotrophy nitrifier, though heterotrophic bacterium has lower decomposition efficiency, but since on its quantity in environment often much larger than autotrophic bacteria, therefore among some environment, the contribution of heterotrophic nitrification effect can be suitable with autotrophic bacteria, even exceed.Some heterotrophic bacterium is nitrification and denitrification simultaneously, makes ammonia in water or nitrous acid after oxidation, can discharge into the atmosphere with the form of gas through denitrogenation, and this kind ability also can provide the competitive edge of this bacterial classification on environment.
At present in the research to nitrification bacteria, fungi is considered to quantity maximum, most effective nitrification bacteria, as Penicillium notatum and fragrant aspergillus tubigensis, Verticillium or the like.In addition, some actinomycetes, bacterium even algae in nature or breadboard pure culture, the heterotrophic microorganism that also is confirmed as having nitrated ability.As denitrifying pseudomonas, Pseudomonas aeruginosa (Pseudomonas aeruginos), Pseudomonas fluorescens (Pseudomonas fluorescens), Alcaligenes, Arthrobacter, Alcaligenes faecalis (Alcaligenus faecalis), PB16 pseudomonas and some newfound bacterial classifications (or genus) or the like.
For effectively regulation and control, monitoring and evaluation nitrification, to the certain research of microbial population distribution carrying out in the denitrification system active sludge, the separation method of the novel reliable separation allotrophic nitrobacteria of setting up of a cover carries out the isolating active discriminating to the allotrophic nitrobacteria in the mixing microorganisms flora in the system and seems very important and necessary, has important theory and using value.
Winogradsky thinks: the autotrophy nitrification can not be grown on beef gelatin flat board, nitrobacteria particularly ammonium salt oxidizing bacteria can only use specific chemoautotrophy mode to live in inorganic salt, can not grow on nutrient gelatin or nutrient agar plate at all.In view of the above in the process of separating the autotrophy nitrobacteria in order whether to have organotrophy microbial contamination in the nutrient solution that confirms to separate the nitrated bacterial strain of autotrophy, will use the meat extract agar flat board tests, it is the auxiliary means that meat extract agar nutrition flat board becomes check " autotrophy nitrobacteria " purity, growth is the assorted bacterium of the nitrated bacterial strain of non-separation purpose autotrophy on it, and never in order to separate the autotrophy nitrobacteria.
The 1950's, Quasel etc. take place to say to traditional nitrification autotrophic bacteria and have proposed query, and are the sole carbon nitrogenous source with the acetoxime, adopt earth-pillar percolation to carry out the selective enrichment cultivation, have obtained to have generation NO
2 -The heterotrophism bacterial strain of ability, investigator afterwards adopts similar methods to carry out the separation of heterotrophic nitrification active bacterial strain more.Substratum commonly used at present has: pyruvic acid oxime, ethanamide and some other organism such as oxime, pyridine etc.The shortcoming of these methods is that sample pre-treatments is more loaded down with trivial details, time and effort consuming, and poor repeatability, strain separated is single and lack representative.
Find by literature search, application number is that 03118598.3 Chinese invention patent discloses a kind of isolation identification purifying heterotrophic nitrification method of microorganism, being characterized as of this patent: after sample to be checked coated beef extract-peptone agar plate, picking list bacterium colony to above-mentioned beef extract-peptone agar plate continues the line purifying, cultivated 10 days through 28 ℃, Griess reagent directly point drips to flat board, carries out nitrification activity and confirms, and make blank with the flat board that does not connect bacterium.Though the operation of the described method of this patent is easy relatively and required time is shorter, so tool is than high practicability, yet its described method has the following disadvantages:
When using aforesaid method, because the not high influence that brings of purity of reagent is difficult for getting rid of, therefore need do high requirement, such as carrying out checking in advance to wherein the nitrite and the content of nitrate to the composition of substratum.Because complicated component in present domestic commercially available peptone and the extractum carnis, if after using the peptone of more easily buying on the market in order to reduce cost, often before the coating and scratching process in the experiment sepn process, periphery of bacterial colonies has a certain amount of nitrite or nitrate with regard to the influence owing to substratum, bring very big influence certainly will for like this judgement of experimental result, thereby have influence on separating resulting allotrophic nitrobacteria.
Summary of the invention
The objective of the invention is to deficiency, propose a kind of method of new separating screening heterotrophic nitration bacteria, simplify the experimental implementation step, reduce in the experiment reagent impurity to the interference of experimental result, thereby improve recall rate at the above various separation method.
For realizing this purpose, the present invention at first prepares extractum carnis-peptone liquid nutrient medium, add the sterilization of agar post-heating, be condensed into the flat-plate solid substratum, then the mixing microorganisms flora is evenly coated extractum carnis-peptone flat board, cultivated 7-10 days down for 30 ℃ in biochemical incubator after leaving standstill condensation, picking list bacterium colony obtains pure bacterium to extractum carnis-peptone plate streaking purifying in the bacterium colony that forms, then pure bacterium lawn is inoculated in ammoxidation capability and identifies nutrient solution, adopt Griess reagent directly to put and splash into and carry out the nitrification activity evaluation in an amount of nutrient solution, qualification test is made blank with the ammoxidation capability evaluation nutrient solution that does not connect the purifying bacterial strain.
Method of the present invention specifically comprises the steps:
1, the preparation of extractum carnis-protein culture medium: press extractum carnis 10g, peptone 10g, NaCl5g, the ratio obtaining liq substratum of 1000mL distilled water, earlier extractum carnis, peptone, NaCl are dissolved in the distilled water, regulating medium pH with the NaOH of 1mol/L was 7.0～8.0 to form liquid nutrient mediums, through 121 ℃ of autoclave sterilizations 15～20 minutes.In above-mentioned extractum carnis-peptone liquid nutrient medium, add 15～20g agar, after fully dissolving it, heating is loaded in the Erlenmeyer flask, sterilized 15～20 minutes down in 121 ℃, treat that substratum is cooled in the culture dish that is poured onto sterilization after 45～50 ℃, leave standstill and be condensed into flat board, obtain extractum carnis-protein culture medium.
2, the dilution of mixing microorganisms flora:
With sample and sterile distilled water is that 1: 9 ratio is put into the container that fills granulated glass sphere by volume, and the vibration dilution of airtight back obtains diluting 10 times sample.Get sample after the dilution and sterile distilled water then in the dilution of vibrating once more of same ratio, carry out altogether 10 times, each extension rate is 10 times, obtains 10 kinds of dilution samples of difference.
3, the coating of mixing microorganisms flora:
From each dilution sample, all get 0.1mL and join respectively on the extractum carnis-protein culture medium flat board for preparing, with the spreading rod of sterilization the dilute sample dispersion is spread out and make its drying.Three repetitions of each dilution sample, extent of dilution series of every variation will be changed transfer pipet again.After leaving standstill condensation, the culture medium flat plate button is come to be put in biochemical incubator cultivated 7-10 days down for 30 ℃.
4, the purifies and separates of bacterium:
In the flat board that forms bacterium colony, pick out the flat board of 90-110 isolated bacterium colony.On the double clean work station of horizontal laminar flow, being housed, the isolated bacterium colony of selecting in the dull and stereotyped culture dish chooses with transfering loop, be transplanted on extractum carnis-protein culture medium, cultivated 7-10 days under 30 ℃ of conditions, then, be put in the sterilized water with transfering loop picking one collarium bacterium lawn again, fully vibration.Use this bacterial suspension, be coated with on flat board according to the viable count measuring method, make it on culture dish, form 30-50 isolated bacterium colony, then isolated bacterium colony is chosen bacterium once according to the method described above again, 2-3 time repeatedly, can obtain the heterotrophism bacterial strain after separation and purification.
5, ammoxidation capability is identified the preparation of nutrient solution:
Ammoxidation capability is identified the preparation of nutrient solution: get glucose 5g/L, (NH
4)
2SO
40.5g/L, NaCl0.3g/L, K
2HPO
41.0g/L, MgSO
4.7H
2O0.3g/L, FeSO
4.7H
2O0.03g/L, CaCO
37.5g/L, being sub-packed in the glass test tube after the thorough mixing dissolving, every test tube packing 3mL uses the silica gel plug jam-pack, in 110 ℃ of following autoclaving 20min, makes ammoxidation capability and identifies nutrient solution.
6, the preparation of Griess reagent:
Take by weighing p-aminophenyl semi-annular jade pendant acid 0.5g, be dissolved in the mixed solution of 30mL Glacial acetic acid and 100mL distilled water, make Sulphanilic Acid solution after the filtration; Take by weighing the 0.1g alpha-naphthylamine and be dissolved in the 100mL hot distilled water, after the cooling, add the 30mL Glacial acetic acid, make alpha-naphthylamine solution after the filtration.Become Griess reagent to use two kinds of solution balanced mix.
7, the nitrification activity checking of liquid pure cultivation:
The heterotrophism bacterial strain that will obtain after separation and purification is inoculated in ammoxidation capability with transfering loop picking one collarium lawn and identifies nutrient solution, under 30 ℃ of conditions, cultivated 7-10 days, got the 2mL nutrient solution to clean tube every 3 days between incubation period, Griess reagent 1-2 is dripped direct point splash into and carry out nitrification activity in the test tube and confirm, and identify that with the ammoxidation capability that does not connect the purifying bacterial strain substratum makes blank.If nutrient solution reddens and can determine that bacterial strain has nitrification activity.
In the process that the single bacterium colony lawn after the separation and purification is moved to ammonia liquor oxidation capacity evaluation nutrient solution, the attention transfering loop don't be touched agar, identifies nutrient solution in order to avoid the impurity in the nutrient agar is brought the ammonia liquor oxidation capacity into, thereby influences qualification result; And when moving into ammoxidation capability evaluation nutrient solution, note not touching test tube wall.
The Griess reagent that is suitable among the present invention need keep in Dark Place in 4 ℃ of refrigerators, use in two weeks, in case find that muddiness need reconfigure.
The present invention separates by extractum carnis-peptone agar plate, ammoxidation capability identify that liquid nutrient medium is cultivated and steps such as Griess reagent colour developing can be from mixing microorganisms be separating screening heterotrophic nitration bacteria, compare with present employed method, have easy, good reproducibility, isolated strains is effective, accurately and reliably characteristics.
Embodiment
Below in conjunction with specific embodiment technical scheme of the present invention is further described.
One embodiment of the present of invention are in the high performance membrane bio-reactor of handling the urban life simulated sewage allotrophic nitrobacteria have been carried out the separation screening process.
Main laboratory apparatus and equipment that method is used have:
The vertical pressure steam sterilizer: the rich Medical Equipment Plant of industry company limited that proves to be true after interrogation in Shanghai produces model YXQ-LS-50SI;
Biochemical incubator: Guangdong Medical Apparatus and Instruments Factory produces, model LRH-250A;
The double clean work station of horizontal laminar flow: Jiangsu Purifying Equipment Co., Ltd., Suzhou produces, model HS-1300;
Constant temperature oscillator: changzhou state China Electrical Appliances Co., Ltd produces model SHZ-82;
Microscope: Japanese Olympus produces, model BX51TF.
Method of the present invention is carried out as follows:
1, the preparation of extractum carnis-protein culture medium:
The prescription of preparation extractum carnis-peptone liquid nutrient medium is as follows: extractum carnis 10g/L, peptone 10g/L, NaCl5g/L.
Making processes: take by weighing extractum carnis 10g, peptone 10g, NaCl5g are dissolved in the 1000mL distilled water, and regulating medium pH with the NaOH of 1mol/L is 7.0-7.2, form liquid nutrient medium, through autoclave sterilization (121 ℃, 15-20min) stand-by.
In above-mentioned extractum carnis-peptone liquid nutrient medium, add 15-20g agar, after fully dissolving it, heating is loaded in the Erlenmeyer flask, in 121 ℃ of 15-20min that sterilize down, treat that substratum is cooled in the culture dish that is poured onto sterilization after 45-50 ℃, leave standstill flat board to be condensed into.Can be placed on dissolving back use in the boiling water during use.
2, the dilution of mixing microorganisms flora:
Draw in the 500mL triangular flask that active sludge 20mL in the membrane bioreactor joins the sterile distilled water that fills 180mL and granulated glass sphere, Mie Jun cotton plug beyond the Great Wall, on constant temperature oscillator, vibrated 10 hours down for 30 ℃ with the 100r/min rotating speed, then getting the 5mL diluent is connected in the 250mL triangular flask that contains 45mL sterile distilled water and granulated glass sphere, Mie Jun silica gel plug beyond the Great Wall, on constant temperature oscillator with 100r/min speed oscillation 10 minutes.This is that for the second time extension rate is 10 dilution, then joins in the sterilized water that 90mL sterilizes with the pipette, extract 10mL of sterilization rapidly.Beyond the Great Wall through the tampon of sterilization, with hand vibration 40 times, get in the sterilized water that 1mL is added to 9mL after dilution finishes for the third time, this is the 4th dilution, then always again according to dilution process serial dilution for the third time 6 times.Carry out 10 dilutions altogether, obtain 10 kinds of dilution samples of difference.
3, the coating of mixing microorganisms flora:
From each dilution sample, all get 0.1mL and join respectively on the extractum carnis-protein culture medium flat board for preparing, with the spreading rod of sterilization the dilute sample dispersion is spread out and make its drying.Three repetitions of each dilution sample, extent of dilution series of every variation will be changed transfer pipet again.After leaving standstill condensation, the culture medium flat plate button is come to be put in biochemical incubator cultivated 7-10 days down for 30 ℃.
4, the purifies and separates of bacterium:
In the flat board that forms bacterium colony, pick out the flat board of 90-110 isolated bacterium colony.On the double clean work station of horizontal laminar flow, the isolated bacterium colony in the culture dish that the flat board of selecting is housed to be chosen with transfering loop, line separates.
Usually, can think that a bacterium colony is come by a cell development, therefore is made up of a kind bacterioid., the possibility that is formed a bacterium colony by diverse several bacterial cells is also arranged.So, need for caution's sake these bacterium are carried out purifying.Behind the bacterium colony to be formed, be transplanted on the solid medium with transfering loop again, cultivated 7-10 days under 30 ℃ of conditions, then, the bacterium lawn of picking one transfering loop is put in the sterilized water again, fully vibration.Use this bacterial suspension, be coated with on flat board according to the viable count measuring method, make it on culture dish, form 30-50 isolated bacterium colony, then isolated bacterium colony is chosen bacterium once according to the method described above again, 2-3 time repeatedly, can obtain the heterotrophism bacterial strain after separation and purification.And do blank with the flat board that does not connect bacterium.
5, ammoxidation capability is identified the preparation of nutrient solution:
It is as follows to fill a prescription: glucose 5g/L, (NH
4)
2SO
40.5g/L, NaCl0.3g/L, K
2HPO
41.0g/L, MgSO
4.7H
2O0.3g/L, FeSO
4.7H
2O0.03g/L, CaCO
37.5g/L.
Making processes: take by weighing above each material respective quality, after the thorough mixing dissolving it is sub-packed in the glass test tube, specification is 18 * 150 millimeters, and every test tube packing 3mL uses the silica gel plug jam-pack, in 110 ℃ of following autoclaving 20min.Make ammoxidation capability and identify nutrient solution.
6, the preparation of Griess reagent:
Griess reagent mixes use by two kinds of solution.1, Sulphanilic Acid solution: take by weighing p-aminophenyl semi-annular jade pendant acid 0.5g, be dissolved in the mixed solution of 30mL Glacial acetic acid and 100mL distilled water, filter, keep in Dark Place, available in this reagent one month; 2, alpha-naphthylamine solution: take by weighing the 0.1g alpha-naphthylamine and be dissolved in the 100mL hot distilled water, after the cooling, add the 30mL Glacial acetic acid, filter, keep in Dark Place, available in this one week of reagent.
Face with preceding two kinds of solution balanced mix.
7, the nitrification activity checking of liquid pure cultivation:
The heterotrophism bacterial strain after separation and purification that obtains is identified substratum by being inoculated in ammoxidation capability with the pure bacterium lawn of transfering loop picking one collarium in extractum carnis-peptone solid medium, cultivated 7-10 days under 30 ℃ of conditions, got the 2mL nutrient solution to clean tube every 3 days between incubation period, Griess reagent 1-2 is dripped direct point splash into and carry out nitrification activity in the test tube and confirm, and identify that with the ammoxidation capability that does not connect the purifying bacterial strain substratum makes blank.As existing redness and even brown, then explanation is owing to nitrification has been put aside nitrite.After 2-3 minute again in the test tube of not colour developing with little spoon add zinc powder a little, leave standstill, whether observation presents redness.As add and show red behind the zinc powder, generation in the nutrient solution then has been described nitrate.Be that bacterial strain also has nitrification activity, tentatively confirm as the nitrification activity bacterium.
Separated the heterotrophic organism with nitrification activity in the mixed bacterial of the present invention by membrane bioreactor system, be numbered B1 and B2 respectively, and carried out relevant bacteriology evaluation and physiological and biochemical test research respectively, the result is as follows:
The bacterium colony that bacterial strain B1 forms after cultivating 1-2 days on the beef-protein medium flat board is creamy white, and is opaque, the Gram-positive genus bacillus, the bacterium size is the μ m of (0.5-0.6) μ m * (1.6-2.6), tool club period of change is not grown for 60 ℃, does not have a resistance to acid.The catalase positive, V-P tests negative, starch hydrolysis feminine gender, the anaerobism agar does not grow, and glucose does not produce acid, except that indivedual physiological and biochemical properties (can utilize glucose to do organic carbon source etc.), its principal character and Bacillus macroides basically identical.
The bacterium colony that bacterial strain B2 forms after cultivating 1 day on the beef-protein medium flat board is creamy white grey, opaque partially, and the bacterium size is the μ m of (0.4-0.6) μ m * (1.2-2.1), tool club period of change.In the gramstaining experiment, bacterial strain is the genus bacillus of being positive at the cultivation initial stage, but the gramstaining instability, and dyeing but is negative about 7 days growing to, and bacterium colony fades to brown.Do not grow for 60 ℃, do not have a resistance to acid.The catalase positive, V-P tests positive, starch hydrolysis feminine gender, the anaerobism agar does not grow, and glucose produces acid, removes indivedual physiological and biochemical properties and (does not have a resistance to acid, on ammonium nitrogen, do not grow) outside, its principal character and bacillus brevis belong to (Brevibacillus) basically identical.
Claims (1)
1, a kind of method of separating screening heterotrophic nitration bacteria is characterized in that comprising the steps:
1) preparation of extractum carnis-protein culture medium: press extractum carnis 10g, peptone 10g, NaCl 5g, the ratio obtaining liq substratum of 1000mL distilled water, earlier extractum carnis, peptone, NaCl are dissolved in the distilled water, regulating medium pH with the NaOH of 1mol/L was 7.0～8.0 to form liquid nutrient mediums, through 121 ℃ of autoclave sterilizations 15～20 minutes; In above-mentioned extractum carnis one peptone liquid nutrient medium, add 15～20g agar, after fully dissolving it, heating is loaded in the Erlenmeyer flask, sterilized 15～20 minutes down in 121 ℃, treat that substratum is cooled in the culture dish that is poured onto sterilization after 45～50 ℃, leave standstill and be condensed into flat board, obtain extractum carnis-protein culture medium;
2) dilution of mixing microorganisms flora: with sample and sterile distilled water is that 1: 9 ratio is put into the container that fills granulated glass sphere by volume, the vibration dilution of airtight back, obtain diluting 10 times sample, get sample after the dilution and sterile distilled water then in the dilution of vibrating once more of same ratio, carry out altogether 10 times, each extension rate is 10 times, obtains 10 kinds of dilution samples of difference;
3) coating of mixing microorganisms flora: from each dilution sample, all get 0.1mL and join respectively on the extractum carnis-protein culture medium flat board for preparing, spreading rod with sterilization disperses dilute sample to spread out to make its drying, three repetitions of each dilution sample, extent of dilution series of every variation, to change transfer pipet again, after leaving standstill condensation, the culture medium flat plate button is come to be put in biochemical incubator cultivated 7-10 days down for 30 ℃;
4) purifies and separates of bacterium: in the flat board that forms bacterium colony, pick out the flat board of 90-110 isolated bacterium colony, on the double clean work station of horizontal laminar flow, being housed, the isolated bacterium colony of selecting in the dull and stereotyped culture dish chooses with transfering loop, be transplanted on extractum carnis-protein culture medium, cultivated 7-10 days under 30 ℃ of conditions, then, be put in the sterilized water with transfering loop picking one collarium bacterium lawn again, fully vibration, on flat board, be coated with according to the viable count measuring method with this bacterial suspension, make it on culture dish, form 30-50 isolated bacterium colony, then isolated bacterium colony is chosen bacterium once according to the method described above again, 2-3 time repeatedly, can obtain the heterotrophism bacterial strain after separation and purification;
5) ammoxidation capability is identified the preparation of nutrient solution: get glucose 5g/L, (NH
4)
2SO
40.5g/L, NaCl0.3g/L, K
2HPO
41.0g/L, MgSO
4.7H
2O0.3g/L, FeSO
4.7H
2O0.03g/L, CaCO
37.5g/L, being sub-packed in the glass test tube after the thorough mixing dissolving, every test tube packing 3mL uses the silica gel plug jam-pack, in 110 ℃ of following autoclaving 20min, makes ammoxidation capability and identifies nutrient solution;
6) preparation of Griess reagent: take by weighing p-aminophenyl semi-annular jade pendant acid 0.5g, be dissolved in the mixed solution of 30mL Glacial acetic acid and 100mL distilled water, make Sulphanilic Acid solution after the filtration; Take by weighing the 0.1g alpha-naphthylamine and be dissolved in the 100mL hot distilled water, after the cooling, add the 30mL Glacial acetic acid, make alpha-naphthylamine solution after the filtration; Become Griess reagent to use two kinds of solution balanced mix;
7) the nitrification activity checking of liquid pure cultivation: the heterotrophism bacterial strain that will obtain after separation and purification is inoculated in ammoxidation capability with transfering loop picking one collarium bacterium lawn and identifies nutrient solution, cultivated 7-10 days under 30 ℃ of conditions, got the 2mL nutrient solution to clean tube every 3 days between incubation period, Griess reagent 1-2 is dripped direct point to splash into and carries out nitrification activity in the test tube and confirm, and identify that with the ammoxidation capability that does not connect the purifying bacterial strain substratum makes blank, if nutrient solution reddens and can determine that bacterial strain has nitrification activity.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101368200B (en) * | 2008-07-18 | 2011-09-21 | 哈尔滨师范大学 | Culture medium for sifting motion anaerobic coreduction sulfate and inverse nitrification function bacterial strain and sifting motion method thereof |
| CN104531595A (en) * | 2015-01-20 | 2015-04-22 | 重庆大学 | Method for rapidly screening and separating heterotrophic nitrification-aerobic denitrification bacterium |
| CN106916873A (en) * | 2017-03-14 | 2017-07-04 | 无锡市中渔健宝生物有限公司 | A kind of method for quick of nitrobacteria nitrification effect |
| CN109517754A (en) * | 2018-11-20 | 2019-03-26 | 上海交通大学 | A method of high temperature bacterial strain is isolated and purified using common biochemical equipment |
| CN114789193A (en) * | 2022-02-17 | 2022-07-26 | 青岛理工大学 | Method for repairing arsenic and antibiotic contaminated soil by bacteria and magnetic diatomite |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1187440C (en) * | 2003-02-14 | 2005-02-02 | 中国科学院南京土壤研究所 | Method for separating, idemtifying, purifying, heterotrophying and nitrifying microbe |
-
2005
- 2005-10-13 CN CNB2005100304563A patent/CN100339487C/en not_active Ceased
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101368200B (en) * | 2008-07-18 | 2011-09-21 | 哈尔滨师范大学 | Culture medium for sifting motion anaerobic coreduction sulfate and inverse nitrification function bacterial strain and sifting motion method thereof |
| CN104531595A (en) * | 2015-01-20 | 2015-04-22 | 重庆大学 | Method for rapidly screening and separating heterotrophic nitrification-aerobic denitrification bacterium |
| CN106916873A (en) * | 2017-03-14 | 2017-07-04 | 无锡市中渔健宝生物有限公司 | A kind of method for quick of nitrobacteria nitrification effect |
| CN109517754A (en) * | 2018-11-20 | 2019-03-26 | 上海交通大学 | A method of high temperature bacterial strain is isolated and purified using common biochemical equipment |
| CN114789193A (en) * | 2022-02-17 | 2022-07-26 | 青岛理工大学 | Method for repairing arsenic and antibiotic contaminated soil by bacteria and magnetic diatomite |
Also Published As
| Publication number | Publication date |
|---|---|
| CN100339487C (en) | 2007-09-26 |
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