CN104388348A - Microbial preparation for purifying sewage and deodorizing garbage and preparation method thereof - Google Patents
Microbial preparation for purifying sewage and deodorizing garbage and preparation method thereof Download PDFInfo
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
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Abstract
The invention provides a microbial preparation for purifying sewage and deodorizing garbage. The microbial preparation is a liquid culture which comprises the following microorganisms: saccharomyces cerevisiae, hansenula polymorpha, rhodopseudomonas palustris, bacillus licheniformis, bacillus subtilis, bacillus megaterium, streptomyces microflavus, lactobacillus acidophilus, lactobacillus plantarum and streptococcus thermophilus. A liquid culturing medium comprises molasses, sodium chloride and urea. The number of viable bacteria in the microbial preparation is more than 1*1010cfu/ml. The microbial preparation can be added into the sewage or sprayed into a garbage transfer station, a garbage landfill and a farm to improve the environment significantly.
Description
Technical field
The invention belongs to probiotics preparing technical field, be specifically related to a kind of probiotics for sewage purification and rubbish deodorizing and preparation method thereof.
Background technology
Probiotics is widely used in the fields such as agricultural, environmental protection, medical treatment, at present agriculturally consumption is maximum, and uses also few at field of Environment Protection.The more existing probiotics for water treatment, although achieve some treatment effects, ubiquity activates the problems such as required time is long, the continuous action time is short, microbe species is single, thus the problem such as cause its treatment effect not good enough.
Summary of the invention
In view of the deficiency of existing field of Environment Protection probiotics, this R&D team inspired by " effective microorganism " concept, prepare a kind of contain multiple-microorganism, can purify waste water and the liquid dosage form probiotics of deodorizing.
A first aspect of the present invention is a kind of probiotics for sewage purification and rubbish deodorizing, it is the liquid culture comprising following microorganism, yeast saccharomyces cerevisiae (Saccharomyces cerevisiae), debaryomyces hansenii (Hansenulaanomala), Rhodopseudomonas palustris (Rhodop seudanonas palustris), Bacillus licheniformis (Bacilluslicheniformis), subtilis [Bacillus subtilis (Ehrenberg) Cohn], bacillus megaterium (Bacillus megaterium), streptomyces microflavus (Streptomyces microflavus), Lactobacterium acidophilum (Lactobacillus acidophilus), plant lactobacillus (Lactobacillus plantarum), thermophilus streptococcus (Streptococcus thermophilus).Wherein cultivation liquid nutrient medium comprises molasses, sodium-chlor, urea, and in probiotics, viable count reaches 1 × 10
10more than cfu/ml.
The present invention also provides a kind of method preparing above-mentioned probiotics, comprising:
(1) prepare composite bacteria, comprise first order seed and cultivate: the bacterium colony of each described microorganism is inoculated in described liquid nutrient medium pure culture 16-24 hour, temperature 30-35 DEG C respectively; And secondary seed is cultivated: one-level culture is inoculated in described liquid culture and amplifies mixed culture 24-48 hour, temperature 30-35 DEG C, obtains the secondary culture as composite bacteria.
(2) Preliminary fermentation: secondary culture is accessed described liquid nutrient medium amplification culture 24-48 hour, temperature 15-30 DEG C; And then add the described liquid nutrient medium of 0.8-1.2 times amount when starting based on Preliminary fermentation, cultivate at 30-35 DEG C, wherein keep pH to be not less than 4.2, illumination is carried out to liquid level, within 4-12 hour, stirs evenly once, be cultured to and occur floating matter.
(3) high sugar-fermenting: the described liquid nutrient medium that additionally with the addition of 3 ~ 5% molasses adding 0.8-1.2 times amount when starting based on Preliminary fermentation again, cultivate at 28 ~ 30 DEG C, pH is wherein kept to be not less than 3.9, continue to provide illumination, within 4-12 hour, stir evenly once, cultivate 4 ~ 8 days, no longer obviously decline to pH;
(4) to mourn in silence fermentation: stop insulation, stir, illumination, when keeping pH 3.9-4.4, continuing to be cultured to viable count and reaching 1 × 10
10more than cfu/mL.
The advantage of probiotics of the present invention is:
Dropped into sewage, as in sanitary sewage, food processing wastewater, livestock-raising waste water, contaminated river course and aquaculture system etc., the indexs such as sewage chemical oxygen demand, total nitrogen, suspended substance can be reduced, thus make water body become limpid; Be sprayed on the places such as garbage transfer station, refuse landfill and plant, the effects such as absorption, enzymolysis can be played to repugnant substances such as the ammonia in air, hydrogen sulfide.Thus the improvement result played environment.
Embodiment
Probiotics of the present invention is obtained through fermentation by composite bacteria.
Composite bacteria preparation and the consisting of of liquid nutrient medium needed for fermentation thereof in the present invention:
Molasses 8 ~ 10%; Salt 1.5 ~ 2%;
Urea 0.5 ~ 1%; Composite growth factor 1 ~ 3%;
And the water of surplus.
Each Ni lead-free solder alloy.Deployed rear sodium hydroxide or sodium bicarbonate adjust pH to 5.8 ~ 6.0, pasteurization.
Above-mentioned molasses, salt, urea are commercial products.
Composite growth factor is autogamy, by 20% dipotassium hydrogen phosphate, 40% magnesium sulfate, 25% filtered tomato juice, 5% yeast extractive substance, 10% electrolysis multidimensional.Wherein, dipotassium hydrogen phosphate, magnesium sulfate, yeast extractive substance, electrolysis multidimensional are commercial products, and filtered tomato juice is self-control.
Composite bacteria is prepared as follows.
The microbial components of composite bacteria comprises yeast saccharomyces cerevisiae (Saccharomyces cerevisiae), debaryomyces hansenii (Hansenula anomala), Rhodopseudomonas palustris (Rhodop seudanonas palustris), Bacillus licheniformis (Bacillus licheniformis), subtilis [Bacillus subtilis (Ehrenberg) Cohn], bacillus megaterium (Bacillus megaterium), streptomyces microflavus (Streptomycesmicroflavus), Lactobacterium acidophilum (Lactobacillus acidophilus), plant lactobacillus (Lactobacillusplantarum), thermophilus streptococcus (Streptococcus thermophilus).Each microbial components is commercial test tube kind.
The preparation method of composite bacteria is divided into one-level and secondary two step.
The pure culture that above-mentioned " one-level " is each component.The bacterium colony of each microorganism of picking, is seeded in 100 ~ 200mL liquid nutrient medium respectively, is placed in constant temperature oscillator 30 ~ 35 DEG C, 100 ~ 200r/min cultivates 16 ~ 24h; But Lactobacterium acidophilum and plant lactobacillus nonoscillatory, quiescent culture.Each microbial inoculant amount is substantially identical.
Above-mentioned " secondary " is mixed culture, concrete grammar is: prepare 5 ~ 10L liquid nutrient medium, yeast saccharomyces cerevisiae, debaryomyces hansenii, Rhodopseudomonas palustris, streptomyces microflavus culture after the cultivation of inoculation one-level, 30 ~ 32 DEG C of intermittent stirring (60 ~ 120r/min mechanical stirring, every 3 ~ 5h once, each 1 ~ 2min) cultivate 12 ~ 24h; Inoculate Bacillus licheniformis, subtilis, bacillus megaterium, Lactobacterium acidophilum, plant lactobacillus, thermophilus streptococcus afterwards, continue cultivation 16 ~ 24h.After cultivation, smelly is considered as dye miscellaneous bacteria, should give up; Smell be tart flavour be normal.
Fermentation culture process is divided into Preliminary fermentation, high sugar-fermenting, fermentation three phases of mourning in silence, and its condition is as described below:
Preliminary fermentation
The composite bacteria prepared is pumped into the seeding tank containing aforesaid liquid substratum, condition ground is not established naturally to cultivate (natural temperature 15 ~ 30 DEG C) 24 ~ 48h, then pump into the fermentor tank that about 1/4th volume of liquid substratum are housed, make culture volume in fermentor tank be about 1/2nd of tank cumulative volume.With 30 ~ 32 DEG C in fermentor tank, pH lower limit 4.2, every 8 ~ 12h stirs 1min, there is provided illumination (such as with high-voltage gas discharging light, as metal halide lamp, xenon lamp etc., irradiate in liquid level on fermentor tank top, act as and promote that Rhodopseudomonas palustris grows and becomes dominant bacteria, usual illumination is not less than 1000LUX; ), be cultured to and occur floating matter (class lawn thing).Usually this stage fermentation time is 1 ~ 3 day.
High sugar-fermenting
In fermentor tank, pump into the described liquid nutrient medium extremely roughly four/three volumes that with the addition of 2 ~ 5% molasses again, stir.With 28 ~ 30 DEG C, pH lower limit 3.9, every 8 ~ 12h stirs 1min, continues to provide illumination, is cultured to pH and no longer obviously declines.Usually this stage fermentation time is 4 ~ 8 days.
To mourn in silence fermentation
Do not reheat, stir, illumination, pH lower limit 3.9, pH the upper limit 4.4 when, continue cultivate, when pH value is lower than lower limit, then add sodium hydroxide or sodium bicarbonate adjustment; When pH value is higher than the upper limit, then add sulfuric acid or glacial acetic acid adjustment.Sink to floating matter major part (more than 80%), and color is bright red or orange-yellow, smell is tart flavour or ester taste, and viable count reaches 1 × 10
10more than cfu/mL can put tank.Usually this stage fermentation time is 5 ~ 10 days.
Below in conjunction with embodiment, the invention will be further described.
Wherein, each embodiment microorganism used therefor bacterial strain and source as follows:
Embodiment 1
1.1 prepare liquid nutrient medium
Get molasses 8%, salt 2%, urea 1%, composite growth factor 2% in mass ratio, add dechlorination tap water and configure 5 tons of substratum.Wherein 2 tons enter 3 tons of seeding tanks, and 2.5 tons enter 10 tons of fermentor tanks, and all the other 0.5 ton for subsequent use.
1.2 prepare composite bacteria
By liquid nutrient medium packing for subsequent use in several 200mL Erlenmeyer flasks, every bottle of 100mL, with 121 DEG C, 15min gives high pressure steam sterilization.After in Bechtop, with transfering loop from some bacterium colonies of picking the test tube kind inclined-plane of each component, be seeded in each bottle substratum, and be placed in constant temperature oscillator 32 DEG C, 150r/min cultivates 24h, but Lactobacterium acidophilum and plant lactobacillus quiescent culture.
Get the liquid nutrient medium that 10L is for subsequent use, inoculation yeast saccharomyces cerevisiae, debaryomyces hansenii, Rhodopseudomonas palustris, streptomyces microflavus, cultivate 24h, between incubation period, stir one minute, stir speed (S.S.) 60r/min with the every 4h of vane type agitator motor for 32 DEG C.Inoculate Bacillus licheniformis, subtilis, bacillus megaterium, Lactobacterium acidophilum, plant lactobacillus, thermophilus streptococcus afterwards also to stir fully, continue quiescent culture 24h.
1.3 Preliminary fermentation
The composite bacteria prepared is pumped into seeding tank, after not establishing condition ground naturally to cultivate 48h, pumps into fermentor tank.With 32 DEG C in fermentor tank, pH lower limit 4.2, every 8h stirs 1min, providing illumination (use metal halide lamp, project, intensity 1000LUX from upper direction liquid level), cultivating 2 days to occurring floating matter.
1.4 high sugar-fermentings
In fermentor tank, pumping into molasses (volume based on newly adding substratum) the extremely about four/three volumes of freshly prepd aforesaid liquid substratum and 5%, stirring.With 28 DEG C, pH lower limit 3.9, every 12h stirs 1min, continues to provide illumination, continues cultivation and no longer obviously declines to pH for 4 days.
1.5 mourn in silence fermentation
Do not reheat, stir, illumination, under the condition of control pH lower limit 3.9, the pH upper limit 4.4, the floating matter continuing to be cultured to 80% sinks, and fermentation system color is bright red or orange-yellow, and smell is tart flavour or ester taste, now stir evenly fermented liquid and sample and carry out enumeration, if viable count reaches 1 × 10
10more than cfu/mL can put tank.
Finally obtain about 8 tons of these probioticses.
Embodiment 2
2.1 prepare liquid nutrient medium
Get molasses 8%, salt 2%, urea 1%, composite growth factor 2% in mass ratio, add dechlorination tap water and configure 10 tons of substratum.Wherein 4 tons enter seeding tank, and 5 tons enter 20 tons of fermentor tanks, and all the other 1 ton for subsequent use.
2.2 prepare composite bacteria
By liquid nutrient medium packing for subsequent use in several 200mL Erlenmeyer flasks, every bottle of 100mL, with 121 DEG C, 15min gives high pressure steam sterilization.After in Bechtop, with transfering loop from some bacterium colonies of picking the test tube kind inclined-plane of each component, be seeded in each bottle substratum, and be placed in constant temperature oscillator 32 DEG C, 150r/min cultivates 24h, Lactobacterium acidophilum and plant lactobacillus quiescent culture.Get the liquid nutrient medium that 10L is for subsequent use, inoculation yeast saccharomyces cerevisiae, debaryomyces hansenii, Rhodopseudomonas palustris, streptomyces microflavus, cultivate 24h, between incubation period, stir one minute, stir speed (S.S.) 60r/min with the every 4h of vane type agitator motor for 32 DEG C.Inoculate Bacillus licheniformis, subtilis, bacillus megaterium, Lactobacterium acidophilum, plant lactobacillus, thermophilus streptococcus afterwards also to stir fully, continue quiescent culture 24h.
2.3 Preliminary fermentation
The composite bacteria prepared is pumped into seeding tank, after not establishing condition ground naturally to cultivate 48h, pumps into fermentor tank.With 32 DEG C in fermentor tank, pH lower limit 4.2, every 8h stirs 1min, provides illumination (use metal halide lamp, from the projection of upper direction liquid level, intensity is about 2000LUX), then cultivates 3 days to occurring floating matter.
2.4 high sugar-fermentings
In fermentor tank, pump into freshly prepd aforesaid liquid substratum and the extremely about four/three volumes of the molasses based on new substratum 5%, stir.With 28 DEG C, pH lower limit 3.9, every 12h stirs 1min, provides illumination, continues cultivation and no longer obviously declines to pH for 5 days.
2.5 mourn in silence fermentation
Do not reheat, stir, illumination, pH lower limit 3.9, the pH upper limit 4.4, be cultured to 80% floating matter sink, culture color is bright red or orange-yellow, and smell is tart flavour or ester taste, and viable count reaches 1 × 10
10more than cfu/mL can put tank.
Finally obtain about 17.5 tons of these probioticses.
using method and effect
(1) embodiment 1 finally obtained evenly being sprayed in certain garbage transfer station with knapsack sprayer after probiotics dilutes 200 times, its deodorizing effect is remarkable, and the change in concentration of ammonia, hydrogen sulfide is as shown in the table:
(2) by the probiotics obtained in embodiment 2 with 0.1% of wastewater influent amount consumption add in the aeration tank of certain food enterprise Sewage treatment systems, once a day, effect is ideal, and change of water quality is as shown in the table: (adding probiotics from August 14)
| Sampling time | Chemical oxygen demand (COD) (COD Cr) | Total nitrogen (TN) | Suspended substance (SS) |
| 8-11 | 187.0 | 32.0 | 71.0 |
| 8-12 | 188.5 | 29.0 | 69.0 |
| 8-13 | 154.5 | 30.5 | 71.0 |
| 8-14 | 161.0 | 21.3 | 69.3 |
| 8-15 | 122.7 | 16.5 | 54.0 |
| 8-16 | 103.0 | 16.8 | 49.0 |
Claims (7)
1. the probiotics for sewage purification and rubbish deodorizing, it is characterized in that, described probiotics is the liquid culture comprising following microorganism: yeast saccharomyces cerevisiae (Saccharomyces cerevisiae), debaryomyces hansenii (Hansenula anomala), Rhodopseudomonas palustris (Rhodop seudanonas palustris), Bacillus licheniformis (Bacillus licheniformis), subtilis [Bacillus subtilis (Ehrenberg) Cohn], bacillus megaterium (Bacillus megaterium), streptomyces microflavus (Streptomycesmicroflavus), Lactobacterium acidophilum (Lactobacillus acidophilus), plant lactobacillus (Lactobacillusplantarum), thermophilus streptococcus (Streptococcus thermophilus),
Wherein microorganism culturing liquid nutrient medium comprises molasses, sodium-chlor, urea, and in probiotics, viable count reaches 1 × 10
10more than cfu/ml.
2. probiotics as claimed in claim 1, described liquid nutrient medium comprises 8 ~ 10% molasses, 1.5 ~ 2% sodium-chlor and 0.5 ~ 1% urea by weight percentage.
3. probiotics as claimed in claim 1, described liquid nutrient medium comprises the water of 8 ~ 10% molasses, 1.5 ~ 2% sodium-chlor, 0.5 ~ 1% urea, 1 ~ 3% composite growth factor and surplus by weight percentage, and pH 5.8 ~ 6.0, wherein composite growth factor comprises 20% dipotassium hydrogen phosphate, 40% magnesium sulfate, 25% filtered tomato juice, 5% yeast extractive substance, 10% electrolysis multidimensional.
4. prepare a method for the probiotics described in any one of claim 1-3, comprising:
(1) composite bacteria is prepared:
First order seed is cultivated: the bacterium colony of each described microorganism is inoculated in described liquid nutrient medium pure culture 16-24 hour, temperature 30-35 DEG C respectively;
Secondary seed is cultivated: one-level culture is inoculated in described liquid culture and amplifies mixed culture 24-48 hour, temperature 30-35 DEG C, obtains the secondary culture as composite bacteria;
(2) Preliminary fermentation: secondary culture is accessed described liquid nutrient medium amplification culture 24-48 hour, temperature 15-30 DEG C; And then add the described liquid nutrient medium of 0.8-1.2 times amount when starting based on Preliminary fermentation, cultivate at 30-35 DEG C, wherein keep pH to be not less than 4.2, illumination is carried out to liquid level, within 4-12 hour, stirs evenly once, be cultured to and occur floating matter;
(3) high sugar-fermenting: the described liquid nutrient medium that additionally with the addition of 3 ~ 5% molasses adding 0.8-1.2 times amount when starting based on Preliminary fermentation again, cultivate at 28 ~ 30 DEG C, pH is wherein kept to be not less than 3.9, continue to provide illumination, within 4-12 hour, stir evenly once, cultivate 4 ~ 8 days, no longer obviously decline to pH;
(4) to mourn in silence fermentation: stop insulation, stir, illumination, when keeping pH 3.9-4.4, continuing to be cultured to viable count and reaching 1 × 10
10more than cfu/mL.
5. preparation method as claimed in claim 4, wherein in first order seed culturing process, the Lactobacterium acidophilum in described microorganism and plant lactobacillus quiescent culture, all the other microorganism shaking culture.
6. preparation method as claimed in claim 4, wherein in secondary seed culturing process, the one-level culture of described Bacillus licheniformis, subtilis, bacillus megaterium, Lactobacterium acidophilum, plant lactobacillus, thermophilus streptococcus is inoculated when the mixed culture time is more than half.
7. prepare a method for the probiotics described in claim 1-3, comprising:
(1) composite bacteria is prepared:
First order seed is cultivated: divided by described liquid nutrient medium and be filled in 200ml Erlenmeyer flask, picking colony from each described microorganism test tube kind inclined-plane, be seeded in each bottle substratum respectively, and be placed in constant temperature oscillator 32 DEG C of shaking culture 24 hours, but Lactobacterium acidophilum and plant lactobacillus quiescent culture;
Secondary seed is cultivated: get liquid nutrient medium described in 10L, the one-level culture of access yeast saccharomyces cerevisiae, debaryomyces hansenii, Rhodopseudomonas palustris, streptomyces microflavus, 32 DEG C of cultivations, within between incubation period every 4 hours, stir once, cultivate and inoculate Bacillus licheniformis, subtilis, bacillus megaterium, Lactobacterium acidophilum, plant lactobacillus, thermophilus streptococcus afterwards in 24 hours, continue quiescent culture after stirring and within 24 hours, obtain secondary culture as composite bacteria;
(2) Preliminary fermentation: secondary culture is accessed 3 tons of described liquid nutrient mediums and cultivate 48 hours, temperature 15-30 DEG C; And then add 2.5 tons of described liquid nutrient mediums, cultivate at 32 DEG C, wherein keep pH to be not less than 4.2, within every 8 hours, stir 1 minute, illumination is provided, is cultured to and occurs floating matter.
(3) high sugar-fermenting: add the described liquid nutrient medium that 3 tons additionally with the addition of 5% molasses again, cultivates at 28 ~ 30 DEG C, wherein keeps pH to be not less than 3.9, continues to provide illumination, within every 12 hours, stirs 1 minute, cultivates 4 ~ 8 days, no longer obviously decline to pH;
(4) to mourn in silence fermentation: stop insulation, stir, illumination, when keeping pH 3.9-4.4, the floating matter continuing to be cultured to 80% sinks, and culture color is bright red or orange-yellow, smell is tart flavour or ester taste, and viable count reaches 1 × more than 1010cfu/mL.
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| CN108217971A (en) * | 2017-12-08 | 2018-06-29 | 广州资源环保科技股份有限公司 | A kind of microbial deodorant and preparation method thereof |
| CN109897798A (en) * | 2019-02-27 | 2019-06-18 | 新兴县国研科技有限公司 | A kind of preparation method and biological deodorant of biological deodorant |
| CN112795514A (en) * | 2021-02-05 | 2021-05-14 | 北京普利赛环保科技发展有限责任公司 | Low-temperature microbial treatment composite powder suitable for dry latrine in cold area and preparation method thereof |
| CN113528386A (en) * | 2021-07-14 | 2021-10-22 | 大连金砣水产食品有限公司 | Preparation method of aquatic product composite microecological preparation |
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| CN109897798A (en) * | 2019-02-27 | 2019-06-18 | 新兴县国研科技有限公司 | A kind of preparation method and biological deodorant of biological deodorant |
| CN112795514A (en) * | 2021-02-05 | 2021-05-14 | 北京普利赛环保科技发展有限责任公司 | Low-temperature microbial treatment composite powder suitable for dry latrine in cold area and preparation method thereof |
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| CN115232763A (en) * | 2022-06-07 | 2022-10-25 | 四川汇邦环保科技有限公司 | Compound microbial agent for separating and conditioning high-density culture water body of grass carp |
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