CN111575200A - Preparation and application method of triple composite deodorant - Google Patents
Preparation and application method of triple composite deodorant Download PDFInfo
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- CN111575200A CN111575200A CN202010374515.3A CN202010374515A CN111575200A CN 111575200 A CN111575200 A CN 111575200A CN 202010374515 A CN202010374515 A CN 202010374515A CN 111575200 A CN111575200 A CN 111575200A
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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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- C02F2101/00—Nature of the contaminant
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- C02F2101/101—Sulfur compounds
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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- 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
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/007—Contaminated open waterways, rivers, lakes or ponds
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Abstract
The invention relates to the technical field of biological environment, in particular to a preparation method of a triple composite deodorant, which consists of thiobacillus thioparus, rhodospirillum sphaeroides and lactobacillus plantarum, and comprises the following specific steps: separating to obtain thiobacillus thioparus, rhodospirillum globosum and lactobacillus plantarum, and storing the strains in an LB solid plate; selecting the three strains by using inoculating loops, and respectively inoculating the three strains into liquid culture media; sequentially carrying out primary fermentation culture on the three strains, and then mixing the three strains according to the proportion of 4: 3: 3 inoculating the mixture into a fermentation tank filled with a liquid culture medium for secondary fermentation culture to obtain the liquid triple composite deodorant. The invention also provides an application method of the triple composite deodorant. The invention organically combines thiobacillus thioparus, lactobacillus and photosynthetic bacteria, can effectively reduce the generation of odorous gas in black and odorous rivers and garbage percolate, and maintains the self-repairing capability of the culture water environment.
Description
Technical Field
The invention relates to the technical field of biological environment, in particular to a preparation method and an application method of a triple composite deodorant.
Background
At present, due to the influence of human activities, the indexes of ammonia nitrogen and sulfide in urban rivers and relatively open water areas are high, and the urban environment is seriously influenced by the black and odorous problem. The resulting ecological environmental problems have been mapped to human home environments and cities. The main sources of the black smelly gas are volatilization of ammonia gas, hydrogen sulfide, methane, thioether, organic acid and other substances. The gas brings inconvenience to the life of residents and has important influence on the health of human bodies.
The existing deodorization technology mainly adopts a chemical method, a physical method and a biological method. The chemical method has the highest deodorization efficiency, but the actual investment cost is the highest, and the secondary pollution is generated; the physical method is mostly covering and can not solve the problem fundamentally. The microbiological method is to decompose and convert odor gas through microbial metabolism from the root, so as to fundamentally achieve the deodorization effect.
The composite microbial agent control scheme has the advantages of simple implementation and capability of removing various black smelly substances. The difficulty in the implementation of the method is how to find the high-efficiency deodorizing strain, maintain the high-efficiency strain with wide adaptation conditions, and how to compound the strain.
Disclosure of Invention
The invention aims to provide a preparation method and an application method of a triple composite deodorant, which aim to solve the problems of screening of high-activity and odor-reducing bacterial strains, compounding of the bacterial strains and application of the bacterial strains in practice.
In order to achieve the purpose, the invention adopts the following technical scheme:
designing a preparation method of a triple composite deodorant, wherein the triple composite deodorant consists of thiobacillus thioparus, rhodospirillum spheroidii and lactobacillus plantarum, and the preparation method comprises the following specific steps:
step 1, strain extraction: separating thiobacillus thioparus from the black and odorous section water body, separating rhodospirillum globosum from the river bottom sludge, separating lactobacillus plantarum from the river bottom sludge, storing the strains in an LB solid flat plate and storing in a refrigerator at 4 ℃;
step 2, strain activation: selecting the three strains in the step 1 by using an inoculating loop, respectively inoculating the three strains into a liquid culture medium, culturing for 18-20h at 30 ℃ and 120rpm by using a shaking table, and storing at 4 ℃ for later use;
step 3, primary fermentation of the strains: sequentially carrying out primary fermentation culture on the three bacteria for standby in the step 2 in the following modes:
rhodospirillum spheroidii culture medium:
2g/l of yeast powder is added,
2.5g/l of sodium acetate,
4g/l of diammonium hydrogen phosphate,
4g/l of sodium chloride is added,
0.5g/l of dipotassium hydrogen phosphate,
magnesium sulfate 0.25g/l, and the culture medium is sterilized at 121 ℃ for 20 minutes;
a thiobacillus thioparus culture medium:
10g/l of sodium chloride is added,
the concentration of the peptone is 10g/l,
1g/l of yeast extract is obtained,
purified water 1L, pH 7.5;
lactobacillus plantarum culture medium:
the concentration of glucose is 60g/l,
30g/l of peptone,
10g/l of yeast powder is added,
3g/l of ammonium citrate is added,
3g/l of anhydrous sodium acetate,
3g/l of dipotassium hydrogen phosphate,
0.2g/l of manganese sulfate,
magnesium sulfate of 0.2g/l,
t-800.1%, the culture temperature is 37 ℃, the pH value is stabilized in the culture process: 5.5, the culture time is 18H, and the inoculation amount is 3 percent;
step 4, secondary fermentation of the strain: compounding the three kinds of strains obtained in step 3, inoculating into a fermentation tank containing liquid culture medium, performing secondary fermentation culture with inoculum size of 5%, and culturing at 30 deg.C for 5 days to obtain final concentration of 108-109CFU/ml, and obtaining the liquid triple composite deodorant.
Furthermore, in step 4, the ratio of thiobacillus thioparvum, rhodospirillum globosum and lactobacillus plantarum is 4: 3: 3.
the invention also provides an application method of the triple composite deodorant, and the triple composite deodorant is sprayed to an area to be deodorized and treated in a spraying mode.
The preparation method and the application method of the triple composite deodorant provided by the invention have the beneficial effects that:
the thiobacillus thioparus exists widely in nature, is harmless to human body, does not pollute environment and has high survival rate. It can decompose and reduce sulfide content in water, and has deodorant effect.
The rhodospirillum spheroidii is a photosynthetic bacterium, can decompose organic matters in a water body, control the nitrogen content and the sulfide content of the water body like many probiotics, has the function of purifying water quality, has no pollution and high efficiency, and is an ideal biological preparation for purifying water quality. The purpose is to achieve the best purification effect with the lowest investment cost.
The invention is based on the view of biological symbiosis, the thiobacillus thioparus, the lactic acid bacteria and the photosynthetic bacteria are organically combined, the thiobacillus and the lactic acid bacteria ensure the rapid reduction of the sulfide and ammonia nitrogen levels of the culture water body, the nitrite reducing function of the photosynthetic bacteria can ensure the stable reduction of the nitrite concentration of the water body, the nitrite content rebound phenomenon possibly caused by the bacillus is eliminated, the synergistic effect of the microorganisms is fully exerted, the defect of a single strain is overcome, the ammonia nitrogen and nitrite nitrogen concentrations in the culture water body are rapidly and efficiently reduced without rebounding, and the self-repairing capability of the culture water body environment is maintained.
Meanwhile, the plant lactic acid bacteria have better capability of secreting protease and amylase, and the photosynthetic bacteria can promote the growth of water animals and can effectively reduce the levels of nitrate and nitrite in the water.
The used strains are obtained by domestication and screening under specific environmental conditions, and the generation of odorous gas in black and odorous rivers and landfill leachate can be effectively reduced.
Detailed Description
The invention is further illustrated by the following examples.
Example 1
A preparation method of a triple composite deodorant, which consists of thiobacillus thioparus, rhodospirillum sphaeroides and lactobacillus plantarum, comprises the following specific steps:
step 1, respectively separating thiobacillus thioparus from water bodies of Chuzhou river black and odorous sections in Liuhe district of Nanjing, separating sludge from river bottoms of urban landscape river of Nanjing to obtain rhodospirillum sphaeroides and lactobacillus plantarum, storing the strains in LB solid plates and storing the strains in a refrigerator at 4 ℃;
and 2, selecting the three strains in the step 1 by using inoculating loops, respectively inoculating the three strains into a liquid culture medium, performing shake culture at 30 ℃ and 120rpm for 18 hours, and storing the three strains at 4 ℃ for later use.
Step 3, primary fermentation of the strains: sequentially carrying out primary fermentation culture on the three bacteria for standby in the step 2 in the following modes:
rhodospirillum spheroidii culture medium: 2g/l of yeast powder, 2.5g/l of sodium acetate, 4g/l of diammonium hydrogen phosphate, 4g/l of sodium chloride, 0.5g/l of dipotassium hydrogen phosphate and 0.25g/l of magnesium sulfate, and sterilizing the culture medium for 20 minutes at 121 ℃;
a thiobacillus thioparus culture medium: 10g/L of sodium chloride, 10g/L of peptone, 1g/L of yeast extract, 1L of purified water and pH 7.5;
lactobacillus plantarum culture medium: 60g/l of glucose, 30g/l of peptone, 10g/l of yeast powder, 3g/l of ammonium citrate, 3g/l of anhydrous sodium acetate, 3g/l of dipotassium phosphate, 0.2g/l of manganese sulfate, 0.2g/l of magnesium sulfate, T-800.1%, the culture temperature is 37 ℃, and the pH value is stabilized in the culture process: 5.5, the culture time is 18H, and the inoculation amount is 3 percent;
step 4, mixing the thiobacillus thioparus, the rhodospirillum globosum and the lactobacillus plantarum obtained in the step 3 according to a ratio of 4: 3: 3 inoculating in a fermentation tank containing liquid culture medium for two-stage fermentation culture with an inoculum size of 5%, and culturing at 30 deg.C for 5 days to obtain final concentration of 108CFU/ml, and obtaining the liquid triple composite deodorant.
Example 2
A preparation method of a triple composite deodorant, which consists of thiobacillus thioparus, rhodospirillum sphaeroides and lactobacillus plantarum, comprises the following specific steps:
step 1, respectively separating thiobacillus thioparus from water bodies of Chuzhou river black and odorous sections in Liuhe district of Nanjing, separating sludge from river bottoms of urban landscape river of Nanjing to obtain rhodospirillum sphaeroides and lactobacillus plantarum, storing the strains in LB solid plates and storing the strains in a refrigerator at 4 ℃;
and 2, selecting the three strains in the step 1 by using inoculating loops, respectively inoculating the three strains into a liquid culture medium, performing shake culture at 30 ℃ and 120rpm for 20 hours, and storing the three strains at 4 ℃ for later use.
Step 3, primary fermentation of the strains: sequentially carrying out primary fermentation culture on the three bacteria for standby in the step 2 in the following modes:
rhodospirillum spheroidii culture medium: 2g/l of yeast powder, 2.5g/l of sodium acetate, 4g/l of diammonium hydrogen phosphate, 4g/l of sodium chloride, 0.5g/l of dipotassium hydrogen phosphate and 0.25g/l of magnesium sulfate, and sterilizing the culture medium for 20 minutes at 121 ℃;
a thiobacillus thioparus culture medium: 10g/L of sodium chloride, 10g/L of peptone, 1g/L of yeast extract, 1L of purified water and pH 7.5;
lactobacillus plantarum culture medium: 60g/l of glucose, 30g/l of peptone, 10g/l of yeast powder, 3g/l of ammonium citrate, 3g/l of anhydrous sodium acetate, 3g/l of dipotassium phosphate, 0.2g/l of manganese sulfate, 0.2g/l of magnesium sulfate, T-800.1%, the culture temperature is 37 ℃, and the pH value is stabilized in the culture process: 5.5, the culture time is 18H, and the inoculation amount is 3 percent;
step 4, mixing the thiobacillus thioparus, the rhodospirillum globosum and the lactobacillus plantarum obtained in the step 3 according to a ratio of 4: 3: 3 inoculating in a fermentation tank containing liquid culture medium for two-stage fermentation culture with an inoculum size of 5%, and culturing at 30 deg.C for 5 days to obtain final concentration of 109CFU/ml, and obtaining the liquid triple composite deodorant.
Example 3
The invention also provides an application method of the triple composite deodorant, which is used for investigating the odor removing effect of the triple microbial inoculum through experiments.
Performing field test in a garbage disposal site at 20-30 deg.C, setting 5 sampling points at the center and periphery of the experimental area, spraying 150ml microbial inoculum according to 1 square meter, detecting the content and degradation rate (average value) of ammonia gas and hydrogen sulfide at 6H, 12H, and 48H, and setting blank control group to examine deodorization effect.
And (3) determining the degradation effect of the triple composite deodorant:
in the early stage of the experiment, the ammonia degradation speed of the experimental group is relatively slow, the ammonia degradation speed is fast after two days, and the degradation rate reaches 79% after 4 days. Wherein, the degradation effect on the hydrogen sulfide is more prominent, and the degradation rate after 48H reaches 88%. And the on-site smell is better improved from the sense of sense. While the control group showed no significant change.
The results show that when thiobacillus thioparus, rhodospirillum sphaeroides and lactobacillus plantarum are mixed according to the proportion of 4: 3: and 3, the residual concentration of ammonia and hydrogen sulfide is obviously reduced along with the prolonging of time.
Example 4
The invention also provides an application method of the triple composite deodorant, which is used for investigating the treatment effect of the triple deodorant on the black and odorous river water through experiments.
The method comprises the steps of carrying out field test in a certain black and odorous river at the temperature of 25 ℃, selecting a certain section of river section with prominent black and odorous, spraying according to the use scheme of spraying 100ml of microbial inoculum by 1 square meter, detecting the content and degradation rate (taking the average value) of ammonia nitrogen and hydrogen sulfide by 12H, 48H and 72H after spraying, and setting a blank control group to investigate the deodorization effect. In the period, the deodorant is added once every 24 hours.
And (3) determining the degradation effect of the triple composite deodorant:
in the early stage of the experiment, the ammonia nitrogen degradation speed of the experimental group is relatively slow, the rapid degradation occurs after two days, and the degradation rate reaches 73% after 3 days. Wherein, the degradation effect on hydrogen sulfide reaches 92.5 percent after 48H.
The results show that when thiobacillus thioparus, rhodospirillum sphaeroides and lactobacillus plantarum are mixed according to the proportion of 4: 3: and 3, the residual concentration of ammonia nitrogen and hydrogen sulfide is obviously reduced along with the prolonging of time.
Proved by experiments, the generation of odorous gas in black and odorous rivers and landfill leachate can be effectively reduced.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (3)
1. The preparation method of the triple composite deodorant is characterized in that the triple composite deodorant consists of thiobacillus thioparus, rhodospirillum spheroidii and lactobacillus plantarum, and the preparation method comprises the following specific steps:
step 1, strain extraction: separating thiobacillus thioparus from the black and odorous section water body, separating rhodospirillum globosum from the river bottom sludge, separating lactobacillus plantarum from the river bottom sludge, storing the strains in an LB solid flat plate and storing in a refrigerator at 4 ℃;
step 2, strain activation: selecting the three strains in the step 1 by using an inoculating loop, respectively inoculating the three strains into a liquid culture medium, culturing for 18-20h at 30 ℃ and 120rpm by using a shaking table, and storing at 4 ℃ for later use;
step 3, primary fermentation of the strains: sequentially carrying out primary fermentation culture on the three bacteria for standby in the step 2 in the following modes:
rhodospirillum spheroidii culture medium:
2g/l of yeast powder is added,
2.5g/l of sodium acetate,
4g/l of diammonium hydrogen phosphate,
4g/l of sodium chloride is added,
0.5g/l of dipotassium hydrogen phosphate,
magnesium sulfate 0.25g/l, and the culture medium is sterilized at 121 ℃ for 20 minutes;
a thiobacillus thioparus culture medium:
10g/l of sodium chloride is added,
the concentration of the peptone is 10g/l,
1g/l of yeast extract is obtained,
purified water 1L, pH 7.5;
lactobacillus plantarum culture medium:
the concentration of glucose is 60g/l,
30g/l of peptone,
10g/l of yeast powder is added,
3g/l of ammonium citrate is added,
3g/l of anhydrous sodium acetate,
3g/l of dipotassium hydrogen phosphate,
0.2g/l of manganese sulfate,
magnesium sulfate of 0.2g/l,
t-800.1%, the culture temperature is 37 ℃, the pH value is stabilized in the culture process: 5.5, the culture time is 18H, and the inoculation amount is 3 percent;
step 4, secondary fermentation of the strain: compounding the three kinds of strains obtained in step 3, inoculating into a fermentation tank containing liquid culture medium, performing secondary fermentation culture with inoculum size of 5%, and culturing at 30 deg.C for 5 days to obtain final concentration of 108-109CFU/ml, and obtaining the liquid triple composite deodorant.
2. The method for preparing a triple composite deodorant according to claim 1, wherein in step 4, the ratio of thiobacillus thioparus, rhodospirillum spheroidii and lactobacillus plantarum is 4: 3: 3.
3. a method of applying a triple composite deodorant according to claim 1, wherein the triple composite deodorant is sprayed in a spraying manner to an area to be deodorized.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4985060A (en) * | 1985-07-04 | 1991-01-15 | Saken Corporation | Soil conditioners |
| CN1563342A (en) * | 2004-02-10 | 2005-01-12 | 凌亮 | Microbe preparation in use for treating high difficult wastewater and preparation method |
| CN103466905A (en) * | 2013-10-10 | 2013-12-25 | 浙江汉蓝环境科技有限公司 | Method for treating nitrobenzene and aniline in wastewater by using microorganisms |
| CN109607820A (en) * | 2018-12-19 | 2019-04-12 | 深圳合民生物科技有限公司 | A kind of water remediation method |
| CN110699283A (en) * | 2019-10-21 | 2020-01-17 | 江苏生久环境科技有限公司 | Biological deodorant for household garbage and preparation method thereof |
-
2020
- 2020-05-06 CN CN202010374515.3A patent/CN111575200A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4985060A (en) * | 1985-07-04 | 1991-01-15 | Saken Corporation | Soil conditioners |
| CN1563342A (en) * | 2004-02-10 | 2005-01-12 | 凌亮 | Microbe preparation in use for treating high difficult wastewater and preparation method |
| CN103466905A (en) * | 2013-10-10 | 2013-12-25 | 浙江汉蓝环境科技有限公司 | Method for treating nitrobenzene and aniline in wastewater by using microorganisms |
| CN109607820A (en) * | 2018-12-19 | 2019-04-12 | 深圳合民生物科技有限公司 | A kind of water remediation method |
| CN110699283A (en) * | 2019-10-21 | 2020-01-17 | 江苏生久环境科技有限公司 | Biological deodorant for household garbage and preparation method thereof |
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