CN109266590A - A kind of denitrification compound bacteria agent and its preparation method and application - Google Patents
A kind of denitrification compound bacteria agent and its preparation method and application Download PDFInfo
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- CN109266590A CN109266590A CN201811422998.9A CN201811422998A CN109266590A CN 109266590 A CN109266590 A CN 109266590A CN 201811422998 A CN201811422998 A CN 201811422998A CN 109266590 A CN109266590 A CN 109266590A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/10—Inorganic compounds
- C02F2101/16—Nitrogen compounds, e.g. ammonia
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/38—Organic compounds containing nitrogen
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Abstract
The invention discloses a kind of denitrification compound bacteria agents and its preparation method and application, and wherein denitrification compound bacteria agent includes complex microorganism and adsorbent；Complex microorganism is made of the bacterium solution of following volumes ratio: Bacillus megatherium 27-32%, thiobacillus denitrificans 18-23%, bacterium nitrobacter 18-23%, nitrosomonas 18-23%, denitrification bacillus 18-23%.The preparation method of denitrification compound bacteria agent of the present invention includes the following steps: that Bacillus megatherium, thiobacillus denitrificans, bacterium nitrobacter, nitrosomonas, denitrification bacillus are carried out to fermented and cultured respectively obtains respective bacterium solution, and mixing is centrifugally separating to obtain complex microorganism by volume, it is mixed again with adsorbent, denitrification compound bacteria agent is obtained after freeze-drying.Denitrification compound bacteria agent prepared by the present invention is strong to environmental suitability, and action effective is fast, and denitrification ability is strong, at low cost, effect is good, environment friendly and pollution-free, and preparation is simple, has positive industrial applicibility.
Description
Technical field
The invention belongs to technical field of sewage, and in particular to a kind of denitrification compound bacteria agent and preparation method thereof and answer
With.
Background technique
As the continuous improvement of the level of urbanization, urban population increasingly expand, industrial or agricultural continues to develop, the rubbish generated
And sewage increasingly increases, and causes environmental pollution, generation bad smell serious, especially water environment pollution accident frequently occurs, seriously
Endanger the health or even life of people, animal.Various synthetics include largely making for chemical fertilizer, pesticide and synthetic detergent etc.
With causing the nitrogenous nutriment of water body to rise, cause water eutrophication, water quality deterioration.It is reported for the nitric wastewater
Denitrogenation technology mainly includes physical method, chemical method and biological method etc., however first two denitrogenation technology be respectively present at
This is higher, treatment effect is bad, easily lead to secondary pollution, the problems such as, and biological prevention and control have broad spectrum activity it is strong, it is at low cost, nontoxic,
The features such as non-corrosive and easy to use is the important means to the nitric wastewater comprehensive treatment, and generally acknowledge relatively safety and
Effective denitrogenation technology.Wherein, Water Pollution Control Engineering is had become using the denitrification treatment technology that microorganism carries out efficiently, economic
One of area research hot spot.
Microbial bacterial agent has been applied in terms of water environment improvement in recent years, but there are still this many problems, such as some
Microorganism cannot generate enzyme, single effect, and rest on laboratory stage more, it is more difficult to be widely applied.In sewerage ecological system,
Single microorganism stability is poor, and denitrification ability is low, bad adaptability, lacks sustainability.Therefore, adaptable, denitrogenation is researched and developed
Ability is strong, and the short composite flora of action effective is most important to the improvement of nitric wastewater.
Summary of the invention
The present invention is intended to provide a kind of pair of denitrification compound bacteria agent and preparation method thereof that environmental suitability is strong, denitrification ability is strong
And application, to effectively solve pollution problem caused by nitric wastewater.
In order to achieve the above objectives, the present invention takes following technical scheme:
Denitrification compound bacteria agent of the present invention, including complex microorganism and adsorbent.
The complex microorganism is made of the bacterium solution of following volumes ratio:
Bacillus megatherium 27-32%, bacterium solution viable count are 1 × 107~5 × 108CFU/mL；
Thiobacillus denitrificans 18-23%, bacterium solution viable count are 1 × 106~2 × 108CFU/mL；
Bacterium nitrobacter 18-23%, bacterium solution viable count are 5 × 106~5 × 108CFU/mL；
Nitrosomonas 18-23%, bacterium solution viable count are 1 × 107~5 × 108CFU/mL；
Denitrification bacillus 18-23%, bacterium solution viable count are 1 × 107~5 × 108CFU/mL。
The adsorbent is mixed to get by turfy soil and diatomite according to the mass ratio of 2-3:8-7.
As a further solution of the present invention: the complex microorganism is mixed by the bacterium solution of following volumes ratio: huge
Bacillus 30%, thiobacillus denitrificans 20%, bacterium nitrobacter 20%, nitrosomonas 20%, denitrification bacillus 10%.It is described
Adsorbent is mixed to get by turfy soil and diatomite according to the mass ratio of 3:7.
The preparation method of denitrification compound bacteria agent of the present invention, includes the following steps:
Step 1: solid slant culture: by Bacillus megatherium, thiobacillus denitrificans, bacterium nitrobacter, nitrosomonas, anti-
Bacterium nitrobacter is transferred on solid medium respectively, and 24~36h is cultivated in 30 DEG C of incubators, activates bacterial strain；
Step 2: seed liquor culture: the bacterial strain activated through step 1 is inoculated in fluid nutrient medium respectively, 30 DEG C, 160~
180rpm 24~36h of shaken cultivation, until bacterial strain is in logarithmic growth phase；
Step 3: expanding culture: the inoculum concentration that the seed liquor that step 2 obtains is 1~5% by volume is inoculated in triangle
In bottle, 30 DEG C, 160~180rpm, 24~36h of shaken cultivation, until bacterial strain is in logarithmic growth phase；
Step 4: collect thallus: the tunning that step 3 is obtained, without living contaminants, is pressed corresponding through pure bacterium experimental verification
Supernatant is removed in volume ratio mixing, centrifugation, obtains concentration thallus；
Step 5: adsorbent sterilizing: turfy soil and diatomite are measured into mixing according to the ratio, 121-150 DEG C heating 1-4 hours
Sterilizing；
Step 6: preparation denitrification compound bacteria agent: the adsorbent after sterilizing being mixed with the concentration thallus ratio of 1:1 in mass ratio
Stirring is closed, dry powder, as denitrification compound bacteria agent is made in -21 DEG C of freeze-dryings.
In step 1, the preparation method of the solid medium is as follows: yeast crude extract 10g, peptone 15g, glucose
5g, NH4Cl 5g, K2HPO45g, agar 20g supplement ddH2O to 1L, pH=7.0,121 DEG C sterilizing 20min.
In step 2 and step 3, the preparation method of the fluid nutrient medium is as follows: yeast crude extract 10g, peptone 15g,
Glucose 5g, NH4Cl 5g, K2HPO45g supplements ddH2O to 1L, pH=7.0,121 DEG C sterilizing 20min.
The use condition of the application method of denitrification compound bacteria agent of the present invention, the denitrification compound bacteria agent requires pH value most preferably
7.0-8.0, optimum temperature are 26-32 DEG C, and every 50kg sewage puts into 0.8-1g denitrification compound bacteria agent of the present invention.
Compared with prior art, it is suitable to environment that the beneficial effects of the present invention are: denitrification compound bacteria agents prepared by the present invention
Ying Xingqiang, action effective is fast, and denitrification ability is strong, at low cost, effect is good, environment friendly and pollution-free, and preparation is simple, has positive industry
Practicability.It is described in detail below:
1, selected microorganism energy efficient denitrification, it is adaptable to Sewage Environment based on the bacterium of sewage ancestral home, it can be quick
Flourish forms dominant microflora, and most of microorganism used in tradition is terrestrial, does not adapt to Sewage Environment, can not be quick
Flourish.
2, selected microorganism can also generate secondary metabolite, such as bacillus megaterium, can be to causing a disease in sewage
Bacterium is inhibited.
3, selected microorganism is composite bacteria agent, has stronger specific aim, combination and cooperative ability, does not repel mutually,
Coordinate mutually in sewage, denitrification effect significantly improves.Complicated technology is not needed when use, is directly launched, it is easy to use.
Specific embodiment
The embodiment of the present invention is described more fully below, is used to explain the present invention, is not considered as limiting the invention.
Embodiment 1:
Denitrification compound bacteria agent of the invention includes following component by weight: complex microorganism: adsorbent=1:1.It is multiple
Closing microorganism includes that bacterium solution by volume mixes: Bacillus megatherium 30%, thiobacillus denitrificans 20%, bacterium nitrobacter
20%, nitrosomonas 20%, denitrification bacillus 10%.Adsorbent is by turfy soil and diatomite according to the mass ratio of 3:7
It is mixed to get.
Each bacterial strain is activated respectively in complex microorganism, culture medium are as follows: yeast crude extract 10g, peptone 15g, grape
Sugared 5g, NH4Cl 5g, K2HPO45g, agar 20g supplement ddH2O to 1L, PH=7.0,121 DEG C sterilizing 20min.Seed liquor training
It supports and expands and cultivate required culture medium are as follows: yeast crude extract 10g, peptone 15g, glucose 5g, NH4Cl 5g, K2HPO45g,
Supplement ddH2O to 1L, PH=7.0,121 DEG C sterilizing 20min.
In complex microorganism each bacterial strain activated respectively, seed liquor culture, expand culture after, by above-mentioned volume ratio progress
After mixing, supernatant is removed in centrifugation, obtains concentration thallus, the adsorbent after sterilizing is mixed according to the above ratio, mixes with concentration thallus
Dry powder, as denitrification compound bacteria agent is made in stirring, frozen drying.
Somewhere sanitary sewage is taken, every 50kg sewage puts into 1g denitrification compound bacteria agent of the present invention, compound not apply the present invention
The sewage of microbial inoculum is blank sample, the ammonia-nitrogen content in sewage is measured respectively with Berthelot spectrophotometry, with alkaline persulfuric acid
Potassium resolution ultraviolet spectrophotometry measures the total nitrogen content in sewage respectively, and degradation rate is calculated on the basis of blank sample, uses this
The degradation rate of invention composite bacteria agent ammonia nitrogen is up to 91.8%, and the degradation rate of total nitrogen is up to 86.9%.
Embodiment 2:
Denitrification compound bacteria agent of the invention includes following component by weight: complex microorganism: adsorbent=1:4.It is multiple
Closing microorganism includes that bacterium solution by volume mixes: Bacillus megatherium 30%, thiobacillus denitrificans 20%, bacterium nitrobacter
20%, nitrosomonas 20%, denitrification bacillus 10%.Adsorbent is by turfy soil and diatomite according to the mass ratio of 2:8
It is mixed to get.
Each bacterial strain is activated respectively in complex microorganism, culture medium are as follows: yeast crude extract 10g, peptone 15g, grape
Sugared 5g, NH4Cl 5g, K2HPO45g, agar 20g supplement ddH2O to 1L, PH=7.0,121 DEG C sterilizing 20min.Seed liquor training
It supports and expands and cultivate required culture medium are as follows: yeast crude extract 10g, peptone 15g, glucose 5g, NH4Cl 5g, K2HPO45g,
Supplement ddH2O to 1L, PH=7.0,121 DEG C sterilizing 20min.
In complex microorganism each bacterial strain activated respectively, seed liquor culture, expand culture after, by above-mentioned volume ratio progress
After mixing, supernatant is removed in centrifugation, obtains concentration thallus, the adsorbent after sterilizing is mixed according to the above ratio, mixes with concentration thallus
Dry powder, as denitrification compound bacteria agent is made in stirring, frozen drying.
Somewhere sanitary sewage is taken, every 50kg sewage puts into 1g denitrification compound bacteria agent of the present invention, compound not apply the present invention
The sewage of microbial inoculum is blank sample, the ammonia-nitrogen content in sewage is measured respectively with Berthelot spectrophotometry, with alkaline persulfuric acid
Potassium resolution ultraviolet spectrophotometry measures the total nitrogen content in sewage respectively, and degradation rate is calculated on the basis of blank sample, uses this
The degradation rate of invention composite bacteria agent ammonia nitrogen is up to 86.7%, and the degradation rate of total nitrogen is up to 79.8%.
Embodiment 3:
Denitrification compound bacteria agent of the invention includes following component by weight: complex microorganism: adsorbent=1:2.It is multiple
Closing microorganism includes that bacterium solution by volume mixes: Bacillus megatherium 30%, thiobacillus denitrificans 20%, bacterium nitrobacter
20%, nitrosomonas 20%, denitrification bacillus 10%.Adsorbent is by turfy soil and diatomite according to the mass ratio of 3:7
It is mixed to get.
Each bacterial strain is activated respectively in complex microorganism, culture medium are as follows: yeast crude extract 10g, peptone 15g, grape
Sugared 5g, NH4Cl 5g, K2HPO45g, agar 20g supplement ddH2O to 1L, PH=7.0,121 DEG C sterilizing 20min.Seed liquor training
It supports and expands and cultivate required culture medium are as follows: yeast crude extract 10g, peptone 15g, glucose 5g, NH4Cl 5g, K2HPO45g,
Supplement ddH2O to 1L, PH=7.0,121 DEG C sterilizing 20min.
In complex microorganism each bacterial strain activated respectively, seed liquor culture, expand culture after, by above-mentioned volume ratio progress
After mixing, supernatant is removed in centrifugation, obtains concentration thallus, the adsorbent after sterilizing is mixed according to the above ratio, mixes with concentration thallus
Dry powder, as denitrification compound bacteria agent is made in stirring, frozen drying.
Somewhere sanitary sewage is taken, every 50kg sewage puts into 0.8g denitrification compound bacteria agent of the present invention, multiple not apply the present invention
The sewage of combined bacteria agent is blank sample, the ammonia-nitrogen content in sewage is measured respectively with Berthelot spectrophotometry, with alkaline over cure
Sour potassium resolution ultraviolet spectrophotometry measures the total nitrogen content in sewage respectively, and degradation rate is calculated on the basis of blank sample, is used
The degradation rate of composite bacteria agent ammonia nitrogen of the present invention is up to 85.9%, and the degradation rate of total nitrogen is up to 78.5%.
Claims (9)
1. a kind of denitrification compound bacteria agent, it is characterised in that: the denitrification compound bacteria agent includes complex microorganism and adsorbent；
The complex microorganism is made of the bacterium solution of following volumes ratio:
Bacillus megatherium 27-32%, bacterium solution viable count are 1 × 107~5 × 108CFU/mL；
Thiobacillus denitrificans 18-23%, bacterium solution viable count are 1 × 106~2 × 108CFU/mL；
Bacterium nitrobacter 18-23%, bacterium solution viable count are 5 × 106~5 × 108CFU/mL；
Nitrosomonas 18-23%, bacterium solution viable count are 1 × 107~5 × 108CFU/mL；
Denitrification bacillus 18-23%, bacterium solution viable count are 1 × 107~5 × 108CFU/mL。
2. denitrification compound bacteria agent according to claim 1, it is characterised in that:
The complex microorganism is mixed by the bacterium solution of following volumes ratio: Bacillus megatherium 30%, thiobacillus denitrificans 20%,
Bacterium nitrobacter 20%, nitrosomonas 20%, denitrification bacillus 10%.
3. denitrification compound bacteria agent according to claim 1, it is characterised in that:
The adsorbent is mixed to get by turfy soil and diatomite according to the mass ratio of 2-3:8-7.
4. denitrification compound bacteria agent according to claim 3, it is characterised in that:
The adsorbent is mixed to get by turfy soil and diatomite according to the mass ratio of 3:7.
5. the preparation method of any denitrification compound bacteria agent in a kind of claim 1-4, it is characterised in that include the following steps:
Step 1: solid slant culture: by Bacillus megatherium, thiobacillus denitrificans, bacterium nitrobacter, nitrosomonas, denitrification
Bacillus is transferred on solid medium respectively, and 24~36h is cultivated in 30 DEG C of incubators, activates bacterial strain；
Step 2: seed liquor culture: the bacterial strain activated through step 1 is inoculated in fluid nutrient medium respectively, 30 DEG C, 160~
180rpm 24~36h of shaken cultivation, until bacterial strain is in logarithmic growth phase；
Step 3: expand culture: the inoculum concentration that the seed liquor that step 2 obtains is 1~5% by volume is inoculated in triangular flask,
30 DEG C, 160~180rpm, 24~36h of shaken cultivation, until bacterial strain is in logarithmic growth phase；
Step 4: collect thallus: the tunning that step 3 is obtained through pure bacterium experimental verification without living contaminants, by corresponding volume
Than mixing, supernatant is removed in centrifugation, obtains concentration thallus；
Step 5: adsorbent sterilizing: measuring mixing for turfy soil and diatomite according to the ratio, sterilizes within heating 1-4 hours at 121-150 DEG C；
Step 6: preparation denitrification compound bacteria agent: the adsorbent after sterilizing being mixed with the concentration thallus ratio of 1:1 in mass ratio and is stirred
It mixes, is freeze-dried, dry powder, as denitrification compound bacteria agent is made.
6. preparation method according to claim 5, it is characterised in that:
In step 1, the preparation method of the solid medium is as follows: yeast crude extract 10g, peptone 15g, glucose 5g,
NH4Cl 5g, K2HPO45g, agar 20g supplement ddH2O to 1L, pH=7.0,121 DEG C sterilizing 20min.
7. preparation method according to claim 5, it is characterised in that:
In step 2 and step 3, the preparation method of the fluid nutrient medium is as follows: yeast crude extract 10g, peptone 15g, grape
Sugared 5g, NH4Cl 5g, K2HPO45g supplements ddH2O to 1L, pH=7.0,121 DEG C sterilizing 20min.
8. the application method of any denitrification compound bacteria agent in a kind of claim 1-4, it is characterised in that:
The use condition of the denitrification compound bacteria agent requires pH value most preferably 7.0-8.0, and optimum temperature is 26-32 DEG C.
9. application method according to claim 8, it is characterised in that:
Every 50kg sewage puts into 0.8-1g denitrification compound bacteria agent of the present invention.
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Cited By (5)
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| CN109576189A (en) * | 2018-12-29 | 2019-04-05 | 合肥市东方美捷分子材料技术有限公司 | A kind of black and odorous water composite bacteria agent and its preparation method and application |
| CN109665677A (en) * | 2019-02-22 | 2019-04-23 | 合肥市东方美捷分子材料技术有限公司 | A kind of terrace-type garbage percolation liquid treating system |
| CN110408565A (en) * | 2019-07-29 | 2019-11-05 | 江苏蓝必盛化工环保股份有限公司 | A kind of efficient denitrification composite bacteria agent, preparation and its application |
| CN112279383A (en) * | 2020-09-29 | 2021-01-29 | 南京乐萱生态环境科技有限公司 | Microbial water purifying agent for rapidly reducing ammonia nitrogen index in water body and preparation method thereof |
| CN114686471A (en) * | 2020-12-30 | 2022-07-01 | 中国石油化工股份有限公司 | Nitrosation microbial inoculum and preparation method thereof |
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| CN112279383A (en) * | 2020-09-29 | 2021-01-29 | 南京乐萱生态环境科技有限公司 | Microbial water purifying agent for rapidly reducing ammonia nitrogen index in water body and preparation method thereof |
| CN114686471A (en) * | 2020-12-30 | 2022-07-01 | 中国石油化工股份有限公司 | Nitrosation microbial inoculum and preparation method thereof |
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