CN106754617A - A kind of method of the low temperature resistant allotrophic nitrobacteria of enrichment culture - Google Patents
A kind of method of the low temperature resistant allotrophic nitrobacteria of enrichment culture Download PDFInfo
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Abstract
The present invention relates to a kind of method of the low temperature resistant allotrophic nitrobacteria of enrichment culture, using industrial wastewater as nutrient solution, enrichment culture is carried out to the activated sludge rich in nitrifier by the way of microbial growth promoters are added, the microbial growth promoters include slaine, polyamines and organic acid azanol, and the slaine is made up of calcium salt, mantoquita, magnesium salts and/or ferrous salt；The domestication of low temperature resistant nitrification bacteria is carried out using low temperature and normal temperature alternately mode, normal temperature cultivation temperature is 18-40 DEG C, and Low- temperature culture temperature is 8-18 DEG C.The present invention is realized by way of adding microbial growth promoters and coordinating normal temperature and low temperature alternate run, the thalline accommodation that is obtained is wide, high treating effect, efficient denitrification can be under cryogenic realized, winter low temperature fluctuation of service can be solved, the problems such as sewage disposal is not up to standard.
Description
Technical field
The invention belongs to field of microbial culture technology, and in particular to a kind of method of the low temperature resistant allotrophic nitrobacteria of enrichment culture.
Background technology
Synchronous nitration and denitrification（SND）Denitrogenation this Process of Biological Nitrogen Removal can simultaneously carry out nitrifying ammonia nitrogen removal and denitrification removing total nitrogen in same reactor, not only overcome some problems of traditional biological denitrification process presence, and have the advantage of protrusion at the aspect such as reducing energy consumption and material consumption：PH stabilizations in reactor can be effectively kept, adding for basicity is reduced or cancel；Reduce the volume of traditional reactor, save capital cost；For the sequencing batch reactor being only made up of a reaction tank, SND can reduce the time needed for realizing nitration denitrification；Save aeration rate, further reducing energy consumption.This technique meets the energy-saving and emission-reduction requirement advocated energetically at present, the study hotspot as sewage treatment area.
Three kinds of explanations, i.e. macro environment explanation, micro theory and biological explanation are formd to synchronous nitration and denitrification biological denitrificaion mechanism at present, they are confirmed and are universally accepted in substantial amounts of research.Either which kind of mechanism, the microorganism for being responsible for denitrogenation is all the Primary Actor for completing biological denitrification process.Since the eighties in 20th century, the research of bioscience man finds many nitrifier such as Pseudomonas fluorescens（Pseudomonas flurescens）, excrement produce bacillus（Alcaligenes facealis）, pseudomonas aeruginosa (Pseudomonas aeruginos) etc. can carry out heterotrophic nitrification to organic or inorganic ammoniate.Compare with autotrophic type nitrifier, although the speed ratio autotrophic bacterium of the heterotroph of biomass oxidation ammonium salt wants the slow 2-3 order of magnitude, but the fast growth of allotrophic nitrobacteria, cell yield are high；It is required that dissolved oxygen concentration is low；Adaptability to environment is also strong, can stand more sour environment, and the speed of the oxidation ammonium salt of its totality is slow unlike autotrophic bacterium.It is exactly under aerobic condition there is denitrifying characteristic to realize simultaneous nitrification and denitrification using certain micro-organisms population that there are researcher in foreign countries.
With the further investigation of synchronous nitration and denitrification technique, the discovery of the nitrification bacteria with aerobic denitrification characteristic theoretically further increases the possibility that sewage nitrification and denitrification is synchronously carried out in a unit, overcome conventional processes in treatment effeciency and economic and practical both sides contradiction simultaneously, realize the denitrogenation of waste water high-efficiency and economy.Efficient degradation bacteria strains are so obtained, the dominant population in reaction system is become and is played denitrification, the process of synchronous nitration and denitrification process industry application will be accelerated.Because aerobic particle mud incubation time is considerably long, formation condition is harsh, hinders the application in industrial production；The characteristic of many microorganisms is not fully understanded that the mechanism of aerobic denitrification and heterotrophic nitrification is not also very clear and definite, to their research is carried out in pure or mixing Bacteria Culture also.So up to the present, only the Sewage Plant such as Holland, Denmark, Italy is run using simultaneous nitrification and denitrification denitrification process, also without large-scale promotion application.Particularly the northern area of China winter because of low temperature effect, the growth and breeding speed and bioactivity of denitrification microorganism can all be greatly lowered in activated sludge, therefore the denitrification effect of many sewage treatment plants is deteriorated, and water outlet ammonia nitrogen concentration is difficult to meet discharge standard.Industrially generally can solve this problem using to cultured low temperature resistant denitrification microorganism is directly delivered in sewage disposal system.
CN201310296747.1 discloses the enrichment and its application in the treatment of waste water of a kind of low temperature resistant heterotrophism synchronous nitration and denitrification microbial inoculum, it is enriched with from laboratory heterotrophic nitrification aerobic denitrifying kind mud, specific method is to carry out culture enrichment and domestication with the ammonia nitrogen containing organic carbon and nitrate nitrogen combined sewage under the conditions of aerobic and two kinds of anoxic, and the heterotrophic nitrification aerobic denitrifying microbial inoculum with low temperature tolerance characteristicses is obtained under the conditions of less than 10 DEG C.The initial ammonia nitrogen concentration of waste water handled by the cultural method only has 36 and 41mg/L, if ammonia nitrogen concentration is higher than the two values in waste water, will be restricted using the method.And the highest denitrification specific degradation rate of the method is 32.93mgNO3-N/(gSS·h）, enrichment time is relatively long.
The content of the invention
In view of the shortcomings of the prior art, the invention provides a kind of method of the low temperature resistant allotrophic nitrobacteria of enrichment culture.The present invention is realized by way of adding microbial growth promoters and coordinating normal temperature and low temperature alternate run, and the thalline accommodation for being obtained is wide, high treating effect, can under cryogenic realize efficient denitrification.After the thalline of enrichment culture of the present invention is added to sewage treatment plant's use, winter low temperature fluctuation of service can be solved, the problems such as sewage disposal is not up to standard.
The method of the low temperature resistant allotrophic nitrobacteria of enrichment culture of the present invention, it is using industrial wastewater as nutrient solution, enrichment culture is carried out to the activated sludge rich in nitrifier by the way of microbial growth promoters are added, the microbial growth promoters include slaine, polyamines and organic acid azanol, and the slaine is made up of calcium salt, mantoquita, magnesium salts and/or ferrous salt；Incubation using low temperature and normal temperature alternately by the way of carry out the domestication of low temperature resistant nitrification bacteria, normal temperature cultivation temperature is 18-40 DEG C, preferably 25-35 DEG C；Low- temperature culture temperature is 8-18 DEG C, preferably 10-15 DEG C.
In microbial growth promoters of the present invention, slaine is 40-100 weight portions, preferably 50-80 weight portions, and polyamines are 5-30 weight portions, preferably 10-20 weight portions, and organic acid azanol is 0.5-15 weight portions, preferably 2-10 weight portions.
In microbial growth promoters of the present invention, slaine can be calcium salt, magnesium salts and mantoquita, wherein Ca2+、Mg2+And Cu2+Mol ratio be（5-15）:（5-25）:（0.5-5）, preferably（8-12）:（10-20）:（1-4）；Or calcium salt, ferrous salt and mantoquita, wherein Ca2+、Fe2+And Cu2+Mol ratio be（5-15）:（1-8）:（0.5-5）, preferably（8-12）:.（2-6）:（1-4）；Or calcium salt, magnesium salts, ferrous salt and mantoquita, wherein Ca2+、Mg2+、Fe2+And Cu2+Mol ratio be（5-15）:（5-25）:（1-8）:（0.5-5）, preferably（8-12）:（10-20）:(2-6):（1-4）.
In microbial growth promoters of the present invention, calcium salt is CaSO4Or CaCl2, magnesium salts is MgSO4Or Mg Cl2, ferrous salt is FeSO4Or FeCl2, mantoquita is CuSO4Or CuCl2。
In microbial growth promoters of the present invention, polyamines are the mixture of spermine, spermidine or both.Organic acid azanol is the mixture of formic acid azanol, hydroxylamine acetate or both.Adding for organic acid azanol can directly participate in the metabolic process of nitrobacteria, shorten enzymatic reaction process, accelerated cell growth as the matrix of azanol oxygen also enzyme.Additionally, the organic acid that organic acid azanol is discharged also helps the growth of some allotrophic nitrobacterias, be conducive to playing the population effect of nitrobacteria, be favorably improved the ability that flora tolerates low temperature environment.
Activated sludge rich in nitrifier of the present invention can be the activated sludge in all aeration tanks commonly used in the art, the optimization treated activated sludge containing ammonia sewage, the inoculum concentration inoculation with 30 minutes settling ratios of sludge as 20%-40%.
The industrial wastewater that the low temperature resistant nitrification bacteria of enrichment culture of the present invention is used includes all of waste water containing ammonia nitrogen and COD：Wherein ammonia nitrogen concentration is generally 50-300mg/L, COD（Cr methods, it is as follows）Concentration is 150-1500mg/L, and pH value is 6.0-9.0.
Can be carbon source commonly used in the art, preferably glucose or acetic acid, more preferably acetic acid in nitrification bacteria Enrichment culture method of the present invention, in addition it is also necessary to periodically add carbon source.Carbon source can disposably be added or flowed and add, according to COD concentration in adding rear nutrient solution for 300-1000mg/L is added.
In nitrification bacteria Enrichment culture method of the present invention, microbial growth promoters can be added once daily or every several days, can also be added when culture medium is changed every time, be added according to promoter concentration 10-50mg/L in cultivating system, preferably 20-40mg/L is added.
Use normal temperature of the present invention culture and Low- temperature culture method alternately, refer to the culture for carrying out certain hour at normal temperatures, preferably carry out the 1-10 culture of batch, accelerate the quick breeding of nitrifier；Then the culture of certain hour is carried out at low temperature, the 1-10 culture of batch is preferably carried out, and tames the low temperature tolerance ability of nitrifier；By normal temperature and the alternate culture of low temperature, enrichment culture obtains low temperature resistant nitrifier.Switch to Low- temperature culture when continuous 3-5 batch nitrogen removal rate is all higher than more than 80% in identical incubation time after changing nutrient solution in preferably middle normal temperature incubation of the invention, normal temperature culture is transferred to when continuous 3-5 batch cell concentration is no longer reduced during Low- temperature culture, this normal temperature and low temperature alternate culture process are repeated 2-6 times, terminate the culture of a cycle, harvest thalline.In the present invention, the dissolved oxygen of normal temperature culture（DO）It is 0.2-1.5mg/L, preferably 0.5-1.0mg/L, the dissolved oxygen of Low- temperature culture is 0.5-3.0mg/L, preferably 1.0-2.0mg/L；Two kinds of pH of cultivating system are 6.0-9.0, preferably 7.8-8.5.
The low temperature resistant nitrification bacteria that the inventive method is enriched with, while having aerobic denitrification function, solves winter sewage disposal plant effluent nitrogen-containing pollutant concentration problem not up to standard after enrichment culture.
Compared with prior art, the present invention has the advantage that：
（1）The microbial growth promoters of special composition and proportioning are added in nitrification bacteria enrichment culture process so that nitrification bacteria realizes fast breeding under the collective effect of slaine, polyamines and organic acid azanol, shortens the enrichment culture time.
（2）By adding microbial growth promoters and coordinating normal temperature and low temperature alternate run in enrichment culture process, enable that low temperature resistant nitrification bacteria rapidly adapts to low temperature environment, being capable of growth and breeding under cryogenic, the good denitrification effect for cooperateing with, improving winter Sewage Plant cold operation can be realized with the activated sludge system in sewage disposal system.
Specific embodiment
The present invention program is described in detail with reference to specific embodiment.
The ratio and formula for being first according to microbial growth promoters shown in table 1 prepare metal salt solution, polyamines and organic acid azanol are added in metal salt solution using preceding, microbial growth promoters I-III are prepared, the promoter concentration is 0.5g/L.
The formula and ratio of the accelerator of table 1
Embodiment 1
According to 20%（30 minutes settling ratios of sludge）Inoculum concentration to accessed in nitrification bacteria cultivation reactor certain sewage treatment plant rich in nitrifier activated sludge, use industry ammonia-containing water water quality for：Ammonia nitrogen concentration is 50-150mg/L, and COD is 50-100mg/L, and pH is 7-8.Microbial growth promoters I are added when nutrient solution is changed per batch, is added according to promoter concentration 30mg/L in cultivating system；Add glucose simultaneously so that the COD concentration in nutrient solution brings up to 300mg/L.Incubation using normal temperature and Low- temperature culture alternately by the way of, first in 25 DEG C of temperature, DO:4 batches of normal temperature culture under conditions of 0.5mg/L, nitrogen removal rate turns 10 DEG C of low temperature, DO when being more than 80%:Cultivated under conditions of 1.0mg/L, continuous 4 batch cell concentrations stabilization, this normal temperature and Low- temperature culture alternately 3 times terminate the incubation of a cycle, harvest low temperature resistant nitrifier A.
Embodiment 2
According to 30%（30 minutes settling ratios of sludge）Inoculum concentration to accessed in nitrification bacteria cultivation reactor certain sewage treatment plant rich in nitrifier activated sludge, use industry ammonia-containing water water quality for：Ammonia nitrogen concentration is：100-200mg/L, COD are that 200-500mg/L, pH are 7.5-8.3.Microbial growth promoters II are added when nutrient solution is changed per batch, is added according to promoter concentration 25mg/L in cultivating system；Add acetic acid simultaneously so that the COD concentration in nutrient solution brings up to 700mg/L.Incubation using normal temperature and Low- temperature culture alternately by the way of, first in 30 DEG C of temperature, DO:4 batches of normal temperature culture under conditions of 0.8mg/L, nitrogen removal rate turns 12 DEG C of low temperature, DO when being more than 80%:Cultivated under conditions of 1.5mg/L, continuous 4 batch cell concentrations stabilization, this normal temperature and Low- temperature culture alternately 4 times terminate the incubation of a cycle, harvest low temperature resistant nitrifier B.
Embodiment 3
According to 40%（30 minutes settling ratios of sludge）Inoculum concentration to accessed in nitrification bacteria cultivation reactor certain sewage treatment plant rich in nitrifier activated sludge, use industry ammonia-containing water water quality for：Ammonia nitrogen concentration is：200-300mg/L, COD are that 500-600mg/L, pH are 7.3-8.5.Microbial growth promoters III are added when nutrient solution is changed per batch, is added according to promoter concentration 20mg/L in cultivating system；Add glucose simultaneously so that the COD concentration in nutrient solution brings up to 900mg/L.Incubation using normal temperature and Low- temperature culture alternately by the way of, first in 35 DEG C of temperature, DO:5 batches of normal temperature culture under conditions of 1.0mg/L, nitrogen removal rate turns 15 DEG C of low temperature, DO when being more than 80%:Cultivated under conditions of 2.0mg/L, continuous 3 batch cell concentrations stabilization, this normal temperature and Low- temperature culture alternately 5 times terminate the incubation of a cycle, harvest low temperature resistant nitrifier C.
Embodiment 4
According to 40%（30 minutes settling ratios of sludge）Inoculum concentration to accessed in nitrification bacteria cultivation reactor certain sewage treatment plant rich in nitrifier activated sludge, use industry ammonia-containing water water quality for：Ammonia nitrogen concentration is：200-300mg/L, COD are that 500-600mg/L, pH are 7.3-8.5.Microbial growth promoters III are added when nutrient solution is changed per batch, is added according to promoter concentration 20mg/L in cultivating system；Add acetic acid simultaneously so that the COD concentration in nutrient solution brings up to 900mg/L.Incubation using normal temperature and Low- temperature culture alternately by the way of, first in 35 DEG C of temperature, DO:5 batches of normal temperature culture under conditions of 1.0mg/L, nitrogen removal rate turns 15 DEG C of low temperature, DO when being more than 80%:Cultivated under conditions of 2.0mg/L, continuous 3 batch cell concentrations stabilization, this normal temperature and Low- temperature culture alternately 5 times terminate the incubation of a cycle, harvest low temperature resistant nitrifier D.
Comparative example 1
Handling process and operating condition with embodiment 1, be a difference in that in incubation not using normal temperature and Low- temperature culture alternately by the way of, temperature is always 25 DEG C.After culture terminates, nitrifier E is harvested.
Comparative example 2
Handling process and operating condition with embodiment 1, be a difference in that in incubation not using normal temperature and Low- temperature culture alternately by the way of, temperature is always 10 DEG C.After culture terminates, nitrifier F is harvested.
Comparative example 3
With operating condition with embodiment 1, difference is handling process：Microbial growth promoters are not added in incubation.After culture terminates, nitrifier G is harvested.
The nitrifier A-G that above-mentioned culture is obtained is used in the processing procedure of certain ammonia-containing water, and wherein ammonia nitrogen concentration is 200mg/L, and COD concentration is 500mg/L, and wastewater processing temperatures are 12 DEG C, and dissolved oxygen concentration is 1-2mg/L, and pH is 7.5-8.5.Within the same reaction time, ammonia nitrogen and nitrogen removal rate are shown in Table 2.
The treatment effect of the low temperature resistant nitrifier of table 2
From the point of view of the data of table 2, low temperature resistant nitrification bacteria A-D under cryogenic Ammonia-Containing Wastewater Treatment when, ammonia nitrogen and nitrogen removal rate are all higher than 80%, only cultivated under normal temperature condition and only cultivate the nitrification bacteria E and F of acquisition under cryogenic, be not added with the nitrification bacteria G of microbial growth promoters acquisition, ammonia nitrogen and nitrogen removal rate are respectively less than 50%.As can be seen here, the culture of low temperature resistant nitrification bacteria must be combined using growth promoter and normal temperature low temperature alternate culture mode, can just be made the thalline of culture and be had good denitrification effect at low temperature.As can be seen here, after the thalline of enrichment culture of the present invention is added to sewage treatment plant's use, winter low temperature fluctuation of service can be solved, the problems such as sewage disposal is not up to standard.
Claims (13)
1. the method for the low temperature resistant allotrophic nitrobacteria of a kind of enrichment culture, it is characterised in that：Using industrial wastewater as nutrient solution, enrichment culture is carried out to the activated sludge rich in nitrifier by the way of microbial growth promoters are added, the microbial growth promoters include slaine, polyamines and organic acid azanol, and the slaine is made up of calcium salt, mantoquita, magnesium salts and/or ferrous salt；The domestication of low temperature resistant nitrification bacteria is carried out using low temperature and normal temperature alternately mode, normal temperature cultivation temperature is 18-40 DEG C, and Low- temperature culture temperature is 8-18 DEG C.
2. method according to claim 1, it is characterised in that：In the microbial growth promoters, slaine is 40-100 weight portions, and polyamines are 5-30 weight portions, and organic acid azanol is 0.5-15 weight portions.
3. method according to claim 1 and 2, it is characterised in that：In the microbial growth promoters, slaine is calcium salt, magnesium salts and mantoquita, wherein Ca2+、Mg2+And Cu2+Mol ratio be（5-15）:（5-25）:（0.5-5）；Or calcium salt, ferrous salt and mantoquita, wherein Ca2+、Fe2+And Cu2+Mol ratio be（5-15）:（1-8）:（0.5-5）；Or calcium salt, magnesium salts, ferrous salt and mantoquita, wherein Ca2+、Mg2+、Fe2+And Cu2+Mol ratio be（5-15）:（5-25）:（1-8）:（0.5-5）.
4. method according to claim 1 and 2, it is characterised in that：In the microbial growth promoters, calcium salt is CaSO4Or CaCl2, magnesium salts is MgSO4Or MgCl2, ferrous salt is FeSO4Or FeCl2, mantoquita is CuSO4Or CuCl2。
5. method according to claim 1 and 2, it is characterised in that：In the microbial growth promoters, polyamines are the mixture of spermine, spermidine or both；Organic acid azanol is the mixture of formic acid azanol, hydroxylamine acetate or both.
6. method according to claim 1, it is characterised in that：The activated sludge rich in nitrifier is the treated activated sludge containing ammonia sewage, the inoculum concentration inoculation with 30 minutes settling ratios of sludge as 20%-40%.
7. method according to claim 1, it is characterised in that：The industrial wastewater is the waste water containing ammonia nitrogen and COD, and wherein ammonia nitrogen concentration is 50-300mg/L, COD（Cr methods, it is as follows）Concentration is 150-1500mg/L, and pH value is 6.0-9.0.
8. method according to claim 1, it is characterised in that：, it is necessary to periodically add carbon source in the allotrophic nitrobacteria Enrichment culture method, according to COD concentration in adding rear nutrient solution for 300-1000mg/L is added.
9. method according to claim 8, it is characterised in that：The carbon source added is glucose or acetic acid.
10. method according to claim 1, it is characterised in that：Microbial growth promoters are daily or added once every several days, or added when culture medium is changed every time, added according to promoter concentration 10-50mg/L in cultivating system.
11. methods according to claim 1, it is characterised in that：Use normal temperature culture and Low- temperature culture method alternately, refer to carry out the 1-10 culture of batch at normal temperatures, accelerate the quick breeding of nitrifier；Then the 1-10 culture of batch is carried out at low temperature, tames the low temperature tolerance ability of nitrifier.
12. methods according to claim 11, it is characterised in that：Switch to Low- temperature culture when continuous 3-5 batch nitrogen removal rate is all higher than more than 80% in identical incubation time after changing nutrient solution in normal temperature incubation, normal temperature culture is transferred to when continuous 3-5 batch cell concentration is no longer reduced during Low- temperature culture, this normal temperature and low temperature alternate culture process are repeated 2-6 times, terminate the culture of a cycle, harvest thalline.
13. method according to claim 1,11 or 12, it is characterised in that：The dissolved oxygen of normal temperature culture is 0.2-1.5mg/L, and the dissolved oxygen of Low- temperature culture is 0.5-3.0mg/L；Two kinds of pH of cultivating system are 6.0-9.0.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111454859A (en) * | 2020-04-07 | 2020-07-28 | 山东海景天环保科技股份公司 | Method for cultivating low-temperature aerobic denitrifying bacteria |
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| CN101899401A (en) * | 2009-05-25 | 2010-12-01 | 中国石油化工股份有限公司 | Microbial agent for treating ammonia-containing waste water and preparation method thereof |
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