CN104152377B - The aerobic denitrifying bacteria of heavy metal tolerance and application thereof - Google Patents
The aerobic denitrifying bacteria of heavy metal tolerance and application thereof Download PDFInfo
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Abstract
A kind of aerobic denitrifying bacteria of heavy metal tolerance of environmental protection technical field and application thereof; does is this bacterial strain bacillus pumilus (Bacillus? pumilus) SNR ?3; be preserved in China General Microbiological culture presevation administrative center (being called for short CGMCC); does is deposit number CGMCC? NO.7126, the present invention can tolerate Cd
2+, Cu
2+in heavy metal, and aerobic denitrification can be carried out under above-mentioned heavy metal existent condition, effectively remove the nitric nitrogen in water body and nitrite nitrogen, and adsorb corresponding heavy metal.This strain culturing condition is extensive, simply easy to operate, and processing cost is low, and the high-efficient purification of the nitrogenous effluent that heavy metal is polluted is significant.
Description
Technical field
What the present invention relates to is a kind of biological treating method of environmental protection technical field, specifically a kind of aerobic denitrifying bacteria of heavy metal tolerance and application thereof.
Background technology
Underground water, as the important component part of water resources, plays a part very important in the evolution of human society.Over nearly 20 years, due to the unreasonable disposal of domestic waste and Industrial " three Waste " etc., a large amount of uses of agriculture production Pesticides, chemical fertilizer, national groundwater pollution situation increases the weight of increasingly.According to the sampling and testing data presentation of Chinese Geological office in 2010, east China main Plain groundwater quality totally causes anxiety, underground water three polluted by nitrogen is general, in facial contamination feature, its exceeding standard rate reaches 7.7 ~ 15.4%, North of Huai River more than 10 province about has 3,000 ten thousand people to drink high nitric acid salt solution, and Haihe basin contaminated underground water stock number accounts for 62% of underground water total resources, and rural area about has 3.6 hundred million people not drink standard compliant tap water.Heavy metal detects generally in point pollution feature, is distributed in the irrigating region around surrounding city and industrial and mining enterprises more.Safe drinking water and people's health have been arrived in the combined pollution serious threat that this multi-pollutant exists jointly.Therefore repair and administer three nitrogen, heavy-metal composite pollution underground water extremely urgent.
Present great majority about the based technique for in-situ remediation of groundwater pollution all for certain first kinds of pollution matter.For the improvement of nitrate pollution, biological denitrificaion is current the most cost-effective Treatment process.Aerobic denitrifying bacteria is the microorganism that a class can play denitrification under aerobic conditions, thus also synchronously can carry out with nitrification, and avoid
to the suppression of nitration reaction, accelerate nitration reaction process.In the method for current process heavy metal wastewater thereby, biosorption process is one of the most promising method.So-called biosorption process is exactly utilize the chemical structure of some organism itself and ingredient properties to adsorb metal ion soluble in water, then removes the method for metal ion in the aqueous solution by solid-liquid two-phase laminated flow.What mention in the report of concerns about bio absorption in recent years is all the adsorption of non-living cells, but the biological adsorption of dead cell is difficult to practical, and for the metal of some kind, dead cell is difficult to active adsorption, and live body bacterium can realize specific adsorption, so people start again to pay close attention to the utilization of living microorganism in the Heavy Metals Bio-adsorption.Metal ion enters viable cell, generally will combine outward through born of the same parents and be transported to two stages in born of the same parents.The former is a Fast Process, does not require the expenditure of energy, is called passive adsorption; The latter carries out comparatively slow, depends on the regulation and control of energy and metabolic system, is called active absorption.Although investigators achieve larger progress at biological denitrificaion and biological adsorption field respectively, also lack the bioremediation technology research that relative synchronous repairs groundwater and heavy metal contamination.
Through finding the retrieval of prior art, open (bulletin) the day 2014.07.23 of Chinese patent literature CN103937701A, disclose a kind of bacillus pumilus shou002 and application thereof, bacillus pumilus (Bacilluspumilus) shou002 is wherein preserved in China typical culture collection center on November 3rd, 2013, deposit number is CCTCCNO:M2013536, this bacillus pumilus shou002 has good restraining effect to aquatic pathogenic bacterium, and no pathogenicity, safe and reliable, can organism effectively in degrading cultivation water, the pollutent such as ammonia nitrogen and cultured water, Optimal culture environment, promote water surrounding benign ecological cycles, can be applied in aquaculture as fodder additives and water ecological setting modifying agent.But this technology does not relate to the degraded of water body nitric nitrogen, and bacterial strain itself does not have the ability of Adsorption of Heavy Metals, cannot be applied to the nitrogenous effluent of heavy metal contamination.
Li Han discloses two strain aerobic denitrifying bacterias in " Nitrogen Removal Mechanism of aerobic denitrifying bacteria and the research of flocculating properties thereof " (Institutes Of Technology Of Nanjing, environmental engineering, 2013, Master's thesis), respectively called after ADB4A and ADB7.Arrange nitric nitrogen influent concentration be 268.8mgL ?1, at 160r/min, cultivate after 2 days under the envrionment conditions of 30 DEG C, strains A DB4A and ADB7 reaches 83% and 63% respectively to the degradation rate of nitric nitrogen.Through 16SrDNA sequencing, think that strains A DB4A and ADB7 is respectively bacillus pumilus and Bacillus licheniformis.In order to understand better and utilize aerobic denitrifying bacteria, the document is set about from factors such as carbon source, carbon-nitrogen ratio, temperature, rotating speed, pH, with single_factor method inquired into aerobic denitrifying bacteria growth optimal environmental condition and environment on the impact of the aerobic denitrification of bacterial strain, all obtain optimum parameter.But this technology is when carrying out denitrification process to water body nitric nitrogen, the accumulation of nitrite nitrogen and ammonia nitrogen in water body can be caused, cause secondary pollution.
Summary of the invention
The present invention is directed to prior art above shortcomings, propose a kind of aerobic denitrifying bacteria and application thereof of heavy metal tolerance, this bacterial strain can tolerate Cd
2+, Cu
2+in heavy metal, and aerobic denitrification can be carried out under above-mentioned heavy metal existent condition, effectively remove the nitric nitrogen in water body and nitrite nitrogen, and adsorb corresponding heavy metal.This strain culturing condition is extensive, simply easy to operate, and processing cost is low, and the high-efficient purification of the nitrogenous effluent that heavy metal is polluted is significant.
The present invention is achieved by the following technical solutions:
The present invention relates to a kind of bacillus pumilus (Bacilluspumilus) SNR ?3, this bacterial strain is preserved in China General Microbiological culture presevation administrative center (being called for short CGMCC) at present, preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, postcode: 100101.Deposit number is CGMCCNO.7126, and preservation date is on January 11st, 2013.
Described bacillus pumilus SNR ?3 there is following character:
1, colonial morphology feature: vegetative cell is the bacillus of 0.4 ~ 0.8 × 1.5 ~ 2.7 μm of sizes on peptone nutrient agar, cultivate for 37 DEG C and form gemma in 2 ~ 3 days, gemma is Long Circle or cylindric.In above-mentioned substratum, 30 DEG C of cultivation 8h thalline can raised growth.Bacterium colony is in white, and there is fold at edge, opaque, does not glisten.
2, physiology and biochemistry characteristic: culture temperature: 20 ~ 37 DEG C, optimum temperuture is 30 DEG C; Grow in the scope of pH5.8 ~ 7.5; Gramstaining tests positive; Methyl red experiment tests positive; Nitrate reduction tests positive; Indoles production testing is positive; H2S production testing is negative; Urase production testing is negative.
Described bacillus pumilus SNR ?3 to record through 16SrDNA sequence, its 16SrDNA major part sequence 1414bp, as shown in SEQIDNo.1, gene order in this sequence and GenBank is carried out tetraploid rice, finds that itself and bacillus pumilus (Bacilluspumilus) homology reach 99.9%.
The present invention relates to a kind of bacillus pumilus (Bacilluspumilus) SNR ?3 application, use it for and remove nitric nitrogen in polluted-water and nitrite nitrogen, be specially: by bacillus pumilus SNR ?after 3 enlarged culturing, be inoculated in containing Cd
2+and/or Cu
2+nitrogenous effluent in, thalline removes nitric nitrogen in water body and nitrite nitrogen by aerobic denitrification in process of growth, utilizes biological adsorption effect to remove Cd in water body simultaneously
2+, Cu
2+heavy metal, and be all positioned at born of the same parents by the heavy metal adsorbed, thus reach the object of removal heavy metal.
Described enlarged culturing refers to: by SNR ?3 inoculation in containing carbon source, nitrogenous source, inorganic salt and water without in bacteria fermentation culture medium, be more than or equal to 5 at C/N ratio, initial pH be 5.8 ~ 7.5, culture temperature be under the environment of 25 ~ 37 DEG C cultivate 6 ~ 24 hours.
Described comprises following component without bacteria fermentation culture medium: carbon source 2 ~ 40g/L, nitrogenous source 0.1 ~ 20g/L, inorganic salt 0.01 ~ 20g/L, all the other are water, pH6.0 ~ 7.5, wherein: described carbon source is any one or a few the combination in glucose, sucrose, maltose, lactose, wood sugar, fructose, citric acid, glycerine and molasses; Nitrogenous source is extractum carnis, peptone, yeast extract paste, starch, corn steep liquor, soybean cake powder, cottonseed meal, urea, (NH
4)
2sO
4, KNO
3and NH4NO
3in any one or a few combination; Inorganic salt are one or more in vitriol, phosphoric acid salt, dihydrogen phosphate and hydrochloride.
Described inoculation refers to: put in nitrogenous effluent according to the ratio of 0.5 ~ 10%.
In described nitrogenous effluent, the starting point concentration of nitric nitrogen is 10 ~ 2000mg/L, nitrite nitrogen starting point concentration is 1 ~ 350mg/L.
Cd in described nitrogenous effluent
2+starting point concentration be 0.01 ~ 10mg/L, Cu
2+starting point concentration be 0.01 ~ 30mg/L.
Technique effect
Compared with prior art, technique effect of the present invention comprises:
1) bacillus pumilus (Bacilluspumilus) that the present invention screens acquisition can utilize several kinds of carbon source and nitrogenous source to carry out growth and breeding, and culture condition is very extensive, simply easy to operate.
2) bacterial strain provided by the invention can carry out denitrification completely under aerobic condition, is nitrogen by nitric nitrogen direct-reduction, realizes thorough denitrogenation and can not cause secondary pollution to environment.
3) bacterial strain provided by the invention has Cd
2+, Cu
2+heavy metal tolerance, can by the biological adsorption effect heavy-metal ion removal of thalline self, and synchronously can carry out with aerobic denitrification process, be applicable to the nitrogenous effluent processing nitrogenous effluent or heavy metal contamination, there is excellent environmental benefit and social benefit.
Accompanying drawing explanation
Fig. 1 be bacillus pumilus (Bacilluspumilus) SNR ?3 colonial morphology photo.
Fig. 2 be bacillus pumilus (Bacilluspumilus) SNR ?3 colony microscope photo (1000 ×).
Embodiment
Elaborate to embodiments of the invention below, the present embodiment is implemented under premised on technical solution of the present invention, give detailed embodiment and concrete operating process, but protection scope of the present invention is not limited to following embodiment.
Embodiment 1
Bacillus pumilus (Bacilluspumilus) SNR ?3 abstraction and purification
Enrichment medium 1: Seignette salt 20g/L, trisodium citrate 10g/L, sodium acetate 10g/L, KNO
32g/L, K
2hPO
40.5g/L, MgSO
47H
2o0.2g/L, trace element solution 0.2%, pH7.5.Trace element solution: EDTA-2Na57.1mg/L, ZnSO
47H
2o3.9mg/L, CaC1
22H
2o7mg/L, MnCl
24H
2o5.1mg/L, FeSO
47H
2o5.0mg/L, CuSO
45H
2o1.6mg/L, CoC1
21.6mg/L.
Enrichment medium 2: add 100mg/LCuSO45H2O on the basis of enrichment medium 1,50mg/L (CH
3cOO)
2cd2H
2o.
BTB substratum: KNO
31.0g/L, C
6h
5na
3o
22H
2o1.0g/L, KH
2pO
41.0/Lg, FeS0
47H
200.05g/L, CaCl
20.2g/L, MgSO
47H
2o1.0g, the BTB1mL of 1%, agar 20.0g/L, pH6.8.
Denitrification substratum: Seignette salt 20g/L, KNO
32.5g/L, K
2hPO
40.5g/L, MgSO
47H
2o0.2g/L, agar 20.0g/L, pH7.5.
Activated sludge sample is gathered from Chongming Island Coastal beach, the activated sludge 10g getting different sampling point respectively adds in enrichment medium 1,30 DEG C, 150r/min cultivates 3d, access in new enrichment medium 1 with 10% inoculum size, continue to cultivate according to the method described above, so repeatedly tame 4 times, with enrichment aerobic denitrifying bacteria.
Above-mentioned enrichment bacterium liquid is accessed in enrichment medium 2 again, carries out acclimating cultivation in the same way, after repeating 4 times, draw lmL enrichment culture liquid in 9mL stroke-physiological saline solution, obtain 10
-1extent of dilution bacteria suspension, is diluted to 10 by 10 times of dilution methods successively
-8.Draw each extent of dilution suspension 0.lmL to BTB solid medium even spread respectively, be inverted cultivation 2 ~ 3d for 30 DEG C, picking has single bacterium colony of blue halos, and on denitrification solid medium, continuous passage cultivated for 6 generations, until obtain the stable pure culture of colonial morphology.
Embodiment 2
Bacillus pumilus (Bacilluspumilus) SNR ?the denitrification effect of 3 pairs of nitric wastewaters
Seed culture medium: glucose 5g/L, extractum carnis 3g/L, peptone 5g/L, NaCl0.5g/L, utilize NaOH solution to adjust pH6.5.
Fermention medium: glucose 20g/L, yeast extract paste 2g/L, peptone 5g/L, K
2hPO
43H
2o2.5g/L, MgSO
47H
2o0.1g/L, utilizes NaOH solution to adjust pH7.0.
By bacillus pumilus B.pumilusSNR ?3 (CGMCCNO.7126) seed culture medium, 30 DEG C, cultivate 10h under the shaking table condition of 200r/min, seed liquor is inoculated in by the grain weight of 2% (v/v) in the 5L fermentor tank of the fermention medium after being preinstalled with 3.5L sterilizing and cultivates, culture condition: 30 DEG C, 300r/min, air flow 1.5L/min, fermenting process is without the need to regulating pH.Fermentation 12h, cell concentration reaches 300 × 10
8cfu/mL.
Above-mentioned bacterium liquid is accessed in the sewage containing nitric nitrogen starting point concentration 300mg/L, nitrite nitrogen starting point concentration 100mg/L by the inoculum size of 4% (v/v) respectively, wherein: C/N ratio is 15.25 DEG C, 300r/min, cultivate 5d under air flow 1.2L/min condition, get nutrient solution centrifugal 10min under 8000rpm condition, get supernatant and detect nitric nitrogen and nitrite nitrogen content.
Table 1 bacterial strain is to the denitrification effect of nitric wastewater
Embodiment 3
Bacillus pumilus (Bacilluspumilus) SNR ?3 couples of Cu
2+the repairing effect of the nitric wastewater polluted
Seed culture medium: glucose 5g/L, extractum carnis 3g/L, peptone 5g/L, MgSO
47H
2o0.3g/L, utilizes NaOH solution to adjust pH6.5.
Fermention medium: glucose 15g/L, sucrose 10g/L, peptone 5g/L, extractum carnis 3g/L, K
2hPO
43H
2o2.5g/L, MgSO
47H
2o0.1g/L, utilizes NaOH solution to adjust pH7.2.
By bacillus pumilus B.pumilusSNR ?3 (CGMCCNO.7126) seed culture medium, 30 DEG C, cultivate 10h under the shaking table condition of 200r/min, seed liquor is inoculated in by the grain weight of 2% (v/v) in the 5L fermentor tank of the fermention medium after being preinstalled with 3.5L sterilizing and cultivates, culture condition: 35 DEG C, 300r/min, air flow 2.0L/min, fermenting process is without the need to regulating pH.Fermentation 8h, cell concentration reaches 300 × 10
8cfu/mL.
Above-mentioned bacterium liquid is contained nitric nitrogen starting point concentration 300mg/L, nitrite nitrogen starting point concentration 50mg/L, Cu by the inoculum size access of 6% (v/v)
2+in the sewage of starting point concentration 5mg/L, wherein C/N ratio is 20,25 DEG C, 300r/min, air flow 1.5L/min time cultivate 5d, get nutrient solution centrifugal 10min under 8000rpm condition, get supernatant detect nitric nitrogen, nitrite nitrogen and Cu
2+content.
Table 2 bacterial strain is to Cu
2+the repairing effect of the nitric wastewater polluted
Embodiment 4
Bacillus pumilus (Bacilluspumilus) SNR ?3 couples of Cd
2+the repairing effect of the nitric wastewater polluted
Seed culture medium: sucrose 5g/L, yeast powder 3g/L, peptone 5g/L, MgSO47H2O0.3g/L, utilize NaOH solution to adjust pH6.8.
Fermention medium: glucose 20g/L, glycerine 5g/L, peptone 5g/L, (NH
4)
2sO
42g/L, K
2hPO
43H
2o2.5g/L, MgSO
47H
2o0.1g/L, utilizes NaOH solution to adjust pH7.2.
By bacillus pumilus B.pumilusSNR ?3 (CGMCCNO.7126) seed culture medium, 30 DEG C, cultivate 10h under the shaking table condition of 200r/min, seed liquor is inoculated in by the grain weight of 4% (v/v) in the 5L fermentor tank of the fermention medium after being preinstalled with 3.5L sterilizing and cultivates, culture condition: 35 DEG C, 400r/min, air flow 1.5L/min, fermenting process is without the need to regulating pH.Fermentation 8h, cell concentration reaches 300 × 10
8cfu/mL.
Above-mentioned bacterium liquid is contained nitric nitrogen starting point concentration 500mg/L, nitrite nitrogen starting point concentration 30mg/L, Cd by the inoculum size access of 6% (v/v)
2+in the sewage of starting point concentration 5mg/L, wherein C/N ratio is 20,25 DEG C, 300r/min, air flow 1.5L/min time cultivate 5d, get nutrient solution centrifugal 10min under 8000rpm condition, get supernatant detect nitric nitrogen, nitrite nitrogen and Cu
2+content.
Table 3 bacterial strain is to Cd
2+the repairing effect of the nitric wastewater polluted
Embodiment 5
Bacillus pumilus (Bacilluspumilus) SNR ?3 couples of Cu
2+, Cd
2+the repairing effect of the nitric wastewater polluted
Seed culture medium: glucose 5g/L, extractum carnis 3g/L, peptone 5g/L, MgSO
47H
2o0.3g/L, utilizes NaOH solution to adjust pH6.5.
Fermention medium: glucose 20g/L, sucrose 10g/L, peptone 5g/L, (NH
4)
2sO
43g/L, K
2hPO
43H
2o2.5g/L, MgSO
47H
2o0.1g/L, utilizes NaOH solution to adjust pH7.2.
By bacillus pumilus B.pumilusSNR ?3 (CGMCCNO.7126) seed culture medium, 30 DEG C, cultivate 10h under the shaking table condition of 200r/min, seed liquor is inoculated in by the grain weight of 4% (v/v) in the 5L fermentor tank of the fermention medium after being preinstalled with 3.5L sterilizing and cultivates, culture condition: 30 DEG C, 400r/min, air flow 1.8L/min, fermenting process is without the need to regulating pH.Fermentation 8h, cell concentration reaches 300 × 10
8cfu/mL.
Above-mentioned bacterium liquid is contained nitric nitrogen starting point concentration 600mg/L, nitrite nitrogen starting point concentration 50mg/L, Cu by the inoculum size access of 10% (v/v)
2+starting point concentration 10mg/L, Cd
2+in the sewage of starting point concentration 2mg/L, wherein C/N ratio is 25,25 DEG C, 300r/min, air flow 1.5L/min time cultivate 5d, get nutrient solution centrifugal 10min under 8000rpm condition,
Get supernatant and detect nitric nitrogen, nitrite nitrogen, Cu
2+, Cd
2+content.
Table 4 bacterial strain is to Cu
2+, Cd
2+the repairing effect of the nitric wastewater polluted
Claims (8)
1. a bacillus pumilus SNR-3, is characterized in that, this bacillus pumilus (
bacilluspumilus) being preserved in China General Microbiological culture presevation administrative center, deposit number is CGMCCNO.7126, and preservation date is on January 11st, 2013; Described bacillus pumilus SNR-3 records sequence as shown in SEQIDNo.1 through 16SrDNA sequence.
2. an application of bacillus pumilus SNR-3 according to claim 1, is characterized in that, uses it for and removes nitric nitrogen in polluted-water and nitrite nitrogen, be specially: after bacillus pumilus SNR-3 enlarged culturing, be inoculated in containing Cd
2+and/or Cu
2+nitrogenous effluent in, thalline removes nitric nitrogen in water body and nitrite nitrogen by aerobic denitrification in process of growth, utilizes biological adsorption effect to remove Cd in water body simultaneously
2+, Cu
2+heavy metal, and be all positioned at born of the same parents by the heavy metal adsorbed, thus reach the object of removal heavy metal.
3. application according to claim 2, it is characterized in that, described enlarged culturing refers to: by SNR-3 inoculation in containing carbon source, nitrogenous source, inorganic salt and water without in bacteria fermentation culture medium, be more than or equal to 5 at C/N ratio, initial pH be 5.8 ~ 7.5, culture temperature be under the environment of 25 ~ 37 DEG C cultivate 6 ~ 24 hours.
4. application according to claim 3, is characterized in that, described comprises following component without bacteria fermentation culture medium: carbon source 2 ~ 40g/L, nitrogenous source 0.1 ~ 20g/L, inorganic salt 0.01 ~ 20g/L, and all the other are water, pH6.0 ~ 7.5.
5. application according to claim 4, is characterized in that, described carbon source is any one or a few the combination in glucose, sucrose, maltose, lactose, wood sugar, fructose, citric acid, glycerine and molasses; Nitrogenous source is extractum carnis, peptone, yeast extract paste, starch, corn steep liquor, soybean cake powder, cottonseed meal, urea, (NH
4)
2sO
4, KNO
3and NH
4nO
3in any one or a few combination; Inorganic salt are one or more in vitriol, phosphoric acid salt or hydrochloride.
6. application according to claim 2, is characterized in that, described inoculation refers to: put in nitrogenous effluent according to the ratio of 0.5 ~ 10%.
7. application according to claim 2, is characterized in that, in described nitrogenous effluent, the starting point concentration of nitric nitrogen is 10 ~ 2000mg/L, nitrite nitrogen starting point concentration is 1 ~ 350mg/L.
8. application according to claim 2, is characterized in that, Cd in described nitrogenous effluent
2+starting point concentration be 0.01 ~ 10mg/L, Cu
2+starting point concentration be 0.01 ~ 30mg/L.
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| CN104862255A (en) * | 2015-05-26 | 2015-08-26 | 浙江大学 | Method for reducing nitrate content of vegetables under protected cultivation conditions |
| CN105950502B (en) * | 2016-05-17 | 2019-09-06 | 浙江大学 | A kind of compound endogenetic bacteria microbial inoculum and its application in heavy-metal contaminated soil phytoremediation |
| CN108977400A (en) * | 2018-09-18 | 2018-12-11 | 宁夏医科大学 | A kind of bacillus pumilus culture medium |
| CN109370954A (en) * | 2018-12-07 | 2019-02-22 | 四川清和科技有限公司 | It is a kind of for the composite bacteria agent of sewage treatment, preparation method and application method |
| CN110357271B (en) * | 2019-06-27 | 2021-09-28 | 杭州秀川科技有限公司 | Rapid biological denitrification method for biochemical effluent of landfill leachate |
| CN111334382B (en) * | 2020-04-17 | 2021-03-12 | 江西理工大学 | Heavy metal cleaning soap based on spores and preparation method thereof |
| CN112811609A (en) * | 2020-12-23 | 2021-05-18 | 福建汇盛生物科技有限公司 | Preparation method of microecological particles for heavy metal adsorption and nitrite degradation |
| CN115259965A (en) * | 2022-07-21 | 2022-11-01 | 广西壮族自治区农业科学院 | Method for realizing paddy soil live selenium and cadmium passivation by utilizing microbial regulation |
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| US6025152A (en) * | 1997-06-03 | 2000-02-15 | Hiatt; William N. | Denitrifying bacterial preparation and method |
| CN1308439C (en) * | 2003-02-14 | 2007-04-04 | 中国科学院南京土壤研究所 | Biological denitrification method for ammonium-nitrogen containing waste water and microbes thereof |
| CN100500831C (en) * | 2003-02-14 | 2009-06-17 | 中国科学院南京土壤研究所 | Denitrifying bacteria composition and its application |
| BRPI0721832B1 (en) * | 2007-08-08 | 2018-03-20 | Peng Guanghao | "METHOD FOR REMOVAL OF CARBON AND NITROGEN CONTAMINANTS FROM WASTE USING HYPEROTROPHIC OXIDANT BACTERIA" |
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