CN102813949B - Microbial environment protectant - Google Patents
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Abstract
The invention discloses a microbial environment protectant, which is characterized by comprising mixed bacteria and a traditional Chinese medicine extract, wherein the mixed bacteria are formed by Rhodopseudomonas palustris, Bacillus Subtilis and Lactobacillus delbruckii. The weight ratio of the Rhodopseudomonas palustris to the Bacillus Subtilis to the Lactobacillus delbruckii is 3:4:7; the volume ratio of the traditional Chinese medicine extract to the mixed bacteria is 1-2 percent. The microbial environment protectant disclosed by the invention has the positive effects that through performing mixed fermentation on the compound bacterial liquid and the traditional Chinese medicine extract, the traditional Chinese medicine ingredients are further activated, and the traditional Chinese medicine ingredients and thallus exoenzyme interact to restrain the incubation of ova, and the generation of adult flies is reduced. As a traditional Chinese medicine and bacterial liquid fermentation medium is special in smell, a trapping and killing effect is performed on the adult flies, and the bad smell of a smelly source can also be effectively prevented from being emitted. The microbial environment protectant disclosed by the invention is harmless to people, is low in cost, simple in preparation process, simple and convenient in operation and remarkable in deodorization and fly killing effect. The problems that mosquitoes and the flies and foul smell exist in refuse landfills, waste transfer stations and digestion tanks are well solved, and a good effect is achieved.
Description
Technical field
The present invention relates to a kind of biological environmental production agent, relate in particular to a kind of deodorization fly eradication cleanser for soot, belong to microbial deodorization field.
Background technology
Along with the fast development of economy, urbanization process is accelerated, and population increases sharply, and the odor pollution problem of house refuse more and more becomes the important environmental problem of government and street levels concern.
Odor pollution belongs to the category of atmospheric pollution.Due to the mixture that stench major part is complicated many combinations, low concentration, the various gaseous matter of low boiling, odorant has the advantages that threshold values is low, sensation intensity is relevant to pollution concentration logarithm, and thus control technology selection aspect will be different from normal atmospheric pollution.Odorant controlled at present has ammonia, hydrogen sulfide, methanthiol, methyl sulfide, trimethylamine, and the polluter producing these stenches lacks effective administering method, does not also have corresponding effectively preventing technology at present to odorant.Main prevention and controls has bioanalysis, absorption method, absorption process, photocatalytic method etc. at present.Multiple technologies combination type of today application is developed into from the operation of single cell processing.(for the dispersion disordered state of China's town and country garbage transfer and landfill, using system equipment, enclosed degraded deodorization, is not solve soot air-polluting main method.) the existing deodorizer of existing market mainly contains chemical deodorizing agent, physical deodorization agent, microbial deodorant.Chemical deodorizing agent is that its cost is high, also can cause secondary pollution by a series of chemical reaction by odorant or neutralization or displacement or be oxidized to odorless material; Physical deodorization agent is absorbing and deodorizing, does not change foul smell combination, only changes the relative concentration of foul smell.Microbial deodorant fly eradication can be current effective method.
The patent of concerns about bio deodorization and document are a lot, and very most of similar to make for the purpose of novel organic fertilizer, therefore flora is numerous and jumbled, namely comprises the bacterial strain producing various enzyme, also needs thermophilic actinomycete, also need the sporeformer of nitrogen-fixing bacteria, dissolving phosphor and dissolving potassium.Although also have deodorizing effect, complex process, production cost is high, and the cycle is long, and effect of killing flies is not good.As Chinese Patent Application No. 200510069596.8, the patent that name is called " a kind of biological deodorization and purification agent and uses thereof ", Chinese Patent Application No. 200510063042.0 for another example, title " method of biological engineering process urban domestic wet refuse ".Another kind of microbial deodorant fly destroying preparation patent, after fermentable completes, the acetic acid lactic acid of volume is added for reducing preparation PH, the culture fluid that hydrogen ion concentration is excessive, microbial enzyme inactivation can be caused, can not normal growth, affect result of use, if Chinese Patent Application No. is 201010124151.X, name is called " refuse landfill deodorization yeast and rubbish deodorization operation technique thereof ", as Chinese Patent Application No. 201110284724.X, title " a kind of microbe lavatory deodorant and preparation method thereof ".
Summary of the invention
The object of this invention is to provide a kind of effective and safe, for the fly eradication maggot of refuse landfill and poultry colony house except the microbial environment protectant of stench, making up the deficiencies in the prior art.
The present invention is the technical scheme realizing object employing:
Microbial environment protectant, is characterized in that: comprise swamp Rhodopseudomonas, bacillus subtilis, lactobacillus delbrueckii formed mixed vaccine and Chinese medicine extraction liquid; By weight, swamp Rhodopseudomonas: bacillus subtilis: lactobacillus delbrueckii=3:4:7; By volume, the ratio of Chinese medicine extraction liquid in mixed vaccine is 1%-2%.
Described swamp Rhodopseudomonas, is prepared from by the following step:
A. make shake-flask seed liquid: by the single bacterium colony of the swamp Rhodopseudomonas through domestication, be inoculated in fluid medium, cultivate 48 hours in 30 DEG C, obtain shake-flask seed liquid;
Described fluid medium is the mixed liquor by following proportions: sodium acetate 0.2 gram, 0.1 gram, sodium chloride, 0.1 gram, magnesium sulfate, potassium dihydrogen phosphate 0.05 gram, ammonium sulphate 0.1 gram, 0.01 gram, calcium chloride, 100ml water, and PH is 6.6; 121 DEG C of sterilizings 30 minutes, cool for subsequent use;
B. production primary seed solution: the swamp Rhodopseudomonas shake-flask seed liquid obtained by step a is with the inoculum concentration of 10% volume ratio, be inoculated into swamp Rhodopseudomonas first order seed liquid culture medium to cultivate 72 hours anaerobic environment 30 DEG C, intensity of illumination is 1000lx, bacterium number reaches 3,000,000,000/ml, obtains swamp Rhodopseudomonas primary seed solution;
Described swamp Rhodopseudomonas first order seed liquid culture medium is the mixed liquor by following proportions: potassium dihydrogen phosphate 1.0 grams, 0.1 gram, calcium chloride, calcium bicarbonate 3 grams, sodium acetate 1 gram, 1.0 grams, sodium chloride, 0.5 gram, magnesium chloride, ammonia chloride 1.0 grams, trace element storage liquid 1ml, growth cofactor storage liquid 1ml, water 1000ml; PH=6.6,121 DEG C of sterilizings 30 minutes, cool for subsequent use;
Described trace element storage liquid is: ferrous chloride 1800 milligrams, cobaltous chloride 250 milligrams, Nickel dichloride. 10 milligrams, copper chloride 10 milligrams, manganese chloride 70 milligrams, zinc chloride 100 milligrams, boric acid 500 milligrams, potassium manganate 30 milligrams, sodium selenate 10 milligrams, water 1000ml;
Described somatomedin storage liquid is: biotin 10 milligrams, nicotinic acid 35 milligrams, thiamine 30 milligrams, para-amino benzoic acid 20 milligrams, 10 milligrams, the many aldehyde of salt acid ratio, calcium pantothenate 10 milligrams, vitamin B
12milligram, water 100ml;
C. fermentation (second order fermentation) is produced: the swamp Rhodopseudomonas primary seed solution obtained by step b is inoculated in fermentation culture with 10% volume ratio, described fermentation culture is same as the first order seed liquid culture medium of step b, condition of culture is same as step b, ferment 96 hours, detect when viable count has reached 3,000,000,000/ml and stop fermentation.
Finally conventional panel count detection is carried out to the viable bacteria content of swamp Rhodopseudomonas fermentation liquid.
Described bacillus subtilis, is prepared from by the following step:
D. make shake-flask seed liquid: by the single bacterium colony of the bacillus subtilis through domestication, be inoculated in fluid medium, cultivate 48 hours in 30 DEG C, obtain shake-flask seed liquid;
Described fluid medium is the mixed liquor by following proportions: peptone 1.0 grams, glucose 1.0 grams, Carnis Bovis seu Bubali cream 1.0 grams, potassium dihydrogen sulfate 0.5 gram, manganese sulfate 0.1 gram, water 1000ml; PH=7.5;
E. production primary seed solution: bacillus subtilis shake-flask seed liquid steps d obtained is with the inoculum concentration of 3% volume ratio, be inoculated into bacillus subtilis first order seed liquid culture medium to cultivate 20 hours anaerobic environment 30 DEG C, bacterium number reaches 3,000,000,000/ml, obtains bacillus subtilis primary seed solution; Bacillus subtilis first order seed liquid culture medium is identical with steps d;
F. produce fermentation (second order fermentation): the bacillus subtilis primary seed solution obtained by step e is inoculated in fermentation culture with 5% volume ratio, described fermentation culture is same as the first order seed liquid culture medium of step e, and condition of culture is same as step e;
Described Lactobacillus delbrueckii, is prepared from by the following step:
G. make shake-flask seed: by the single colony inoculation of moral formula lactobacillus through domestication in Deshi Lactobacillus fluid medium, cultivate 36 hours, obtain shake-flask seed liquid for 37 DEG C;
Described fluid medium is the mixed liquor by following proportions: yeast extract 5 grams, peptone 5 grams, glucose 9 grams, Tween 80 0.5 gram, potassium dihydrogen phosphate 2 grams, Fructus Lycopersici esculenti juice 100ml, water 1000ml; PH=5.6,121 DEG C of sterilizings 15 minutes.
H. production primary seed solution: lactobacillus delbrueckii seed liquor step g obtained, with the inoculum concentration of 5% volume ratio, is inoculated in first order seed liquid culture medium, cultivates 40 hours in 45 DEG C, reaches 1,000,000,000/ml, obtain primary seed solution to viable count.
Described first order seed liquid culture medium prescription is the mixed liquor by following proportions: sucrose 10 grams, 25 grams, molasses, peptone 5 grams, yeast extract 5 grams, 0.5 gram, magnesium sulfate, 0.1 gram, sodium chloride, potassium dihydrogen phosphate 0.5 gram, water 1000ml;
I. produce fermentation: the seed liquor obtained by step h, with the inoculum concentration of 15% volume ratio, is inoculated in fermentation liquid culture medium, cultivate 48 hours in 40 DEG C, when dropping to 1 gram of L with residual sugar, fermentation ends;
Described fermentation liquid culture medium prescription is the mixed liquor by following proportions, 40 grams, molasses, sucrose 20 grams, wheat root 18 grams, peptone 5 grams, yeast extract 5 grams, 0.5 gram, magnesium sulfate, 0.1 gram, sodium chloride, potassium dihydrogen phosphate 0.5 gram, water 1000ml;
Described extracts of Chinese herbal medicine, be prepared from by the following step: Chinese medicine Herba polygoni hydropiperis, the Radix Euphorbiae Fischerianae (Radix Euphorbiae Ebracteolatae), Radix Clematidis, northern Cortex Dictamni, Herba Xanthii are mixed according to weight ratio 5:3:1:1:1, add the alcohol-pickled 24-72 hour of 4 ° of 10 times of volumes, again the medicine after immersion is decocted 20-30 minute under boiling water condition, it is stand-by that drop goes out medicinal liquid.
Described mixed vaccine, is prepared from by the following step:
By weight, swamp Rhodopseudomonas: bacillus subtilis: join in mixed bacteria liquid culture medium after lactobacillus delbrueckii=3:4:7 mixing and ferment 15 days, viable count reaches 1,000,000,000/ml, and during PH<4 prepared by mixed liquor;
Described multi strain co cultivation base is be the mixed liquor by following proportions: molasses 30%, magnesium sulfate 1%, ferrous sulfate 0.4%, and potassium dihydrogen phosphate 1%, boils and put to room temperature;
Good effect of the present invention is: the present invention utilizes multiple-microorganism and its metabolite, to the domestic pollution source producing stench in city, i.e. ammonia, hydrogen sulfide, indole, methanthiol, aldehydes in refuse transfer station and Treatment stations, carries out absorption degraded conversion, to reach the object of deodorization.Wherein bacillus subtilis cell mass, Drug resistance is strong, and produces protease and chitinase, effective decomposition insect chorion and fungal cell wall.Bacillus subtilis is a kind of Nitrite transformation bacterium, the ammoniacal nitrogen of rubbish can be converted into nitroso-group nitrogen, and nitroso-group nitrogen transformation is nitrate, and nitrate can be absorbed by plants.Photosynthetic bacteria utilizes nitrate to form tropina, thus reduces the generation of ammoniacal nitrogen, reduces the content of ammonia.Photosynthetic bacteria is that Rhodopseudomonas belongs to, and utilize the ability of sulfide to increase through domestication, hydrogen sulfide can be used as oxygen donator, and ammoniacal nitrogen can be utilized to form drive member.Deshi Lactobacillus lactic acid producing is very capable, Ke Yi Shi ?the acidic liquid of liquid PH<4.The metabolism of Absorbable rod stink substance and suppression putrefaction bacteria and breeding.By the comprehensive function of microorganism and metabolite thereof, to the original transfer capability of degrading more by force of foul smell, reach obvious deodorizing effect.In addition, the vitamin of Chinese medicine extraction liquid promotes mixed bacteria liquid growth, and the enzyme in mixed bacteria liquid guards against enzyme etc. as soap can make the secondary metabolite of plant such as alkaloid become active higher material from activity is lower, thus improves drug effect.Summary: the present invention, by composite bacteria liquid and extracts of Chinese herbal medicine mixed culture fermentation, impels Chinese medicine ingredients to activate further, suppresses the hatching of worm's ovum, decrease into the generation of fly with the combined effect of thalline spore exoenzyme.Because Chinese herbal medicine and bacterium liquid fermentation medium have special odor, have trapping effect to one-tenth fly, the foul smell that also effectively can intercept smelly source distributes.Microbial environment protectant of the present invention is harmless, and cost is low, and preparation technology is simple, easy and simple to handle, deodorization effect of killing flies is remarkable, preferably resolves refuse landfill, garbage transfer station and lavatory, the mosquitos and flies of anaerobic tank and malodor problem, reaches good deodorization effect of killing flies.
Accompanying drawing explanation
The implementation result table of Fig. 1 embodiment of the present invention.
Detailed description of the invention
Embodiment 1:
the preparation of swamp Rhodopseudomonas (pseudmonas palustris) bacterium:
A. make shake-flask seed liquid: by the single bacterium colony of the swamp Rhodopseudomonas through domestication, be inoculated in fluid medium, cultivate 48 hours in 30 DEG C, obtain shake-flask seed liquid;
Described fluid medium is the mixed liquor by following proportions: sodium acetate 0.2 gram, 0.1 gram, sodium chloride, 0.1 gram, magnesium sulfate, potassium dihydrogen phosphate 0.05 gram, ammonium sulphate 0.1 gram, 0.01 gram, calcium chloride, 100ml water, and PH is 6.6; 121 DEG C of sterilizings 30 minutes, cool for subsequent use;
B. production primary seed solution: the swamp Rhodopseudomonas shake-flask seed liquid obtained by step a is with the inoculum concentration of 10% volume ratio, be inoculated into swamp Rhodopseudomonas first order seed liquid culture medium to cultivate 72 hours anaerobic environment 30 DEG C, intensity of illumination is 1000lx, bacterium number reaches 3,000,000,000/ml, obtains swamp Rhodopseudomonas primary seed solution;
Described swamp Rhodopseudomonas first order seed liquid culture medium is the mixed liquor by following proportions: potassium dihydrogen phosphate 1.0 grams, 0.1 gram, calcium chloride, calcium bicarbonate 3 grams, sodium acetate 1 gram, 1.0 grams, sodium chloride, 0.5 gram, magnesium chloride, ammonia chloride 1.0 grams, trace element storage liquid 1ml, growth cofactor storage liquid 1ml, water 1000ml; PH=6.6,121 DEG C of sterilizings 30 minutes, cool for subsequent use;
Described trace element storage liquid is: ferrous chloride 1800 milligrams, cobaltous chloride 250 milligrams, Nickel dichloride. 10 milligrams, copper chloride 10 milligrams, manganese chloride 70 milligrams, zinc chloride 100 milligrams, boric acid 500 milligrams, potassium manganate 30 milligrams, sodium selenate 10 milligrams, water 1000ml;
Described somatomedin storage liquid is: biotin 10 milligrams, nicotinic acid 35 milligrams, thiamine 30 milligrams, para-amino benzoic acid 20 milligrams, 10 milligrams, the many aldehyde of salt acid ratio, calcium pantothenate 10 milligrams, vitamin B
12milligram, water 100ml;
C. fermentation (second order fermentation) is produced: the swamp Rhodopseudomonas primary seed solution obtained by step b is inoculated in fermentation culture with 10% volume ratio, described fermentation culture is same as the first order seed liquid culture medium of step b, condition of culture is same as step b, ferment 96 hours, detect when viable count has reached 3,000,000,000/ml and stop fermentation.
Finally conventional panel count detection is carried out to the viable bacteria content of swamp Rhodopseudomonas fermentation liquid.
The preparation of bacillus subtilis (Rhodo pseudmonas palustris):
D. make shake-flask seed liquid: by the single bacterium colony of the bacillus subtilis through domestication, be inoculated in fluid medium, cultivate 48 hours in 30 DEG C, obtain shake-flask seed liquid;
Described fluid medium is the mixed liquor by following proportions: peptone 1.0 grams, glucose 1.0 grams, Carnis Bovis seu Bubali cream 1.0 grams, potassium dihydrogen sulfate 0.5 gram, manganese sulfate 0.1 gram, water 1000ml; PH=7.5;
E. production primary seed solution: bacillus subtilis shake-flask seed liquid steps d obtained is with the inoculum concentration of 3% volume ratio, be inoculated into bacillus subtilis first order seed liquid culture medium to cultivate 20 hours anaerobic environment 30 DEG C, bacterium number reaches 3,000,000,000/ml, obtains bacillus subtilis primary seed solution; Bacillus subtilis first order seed liquid culture medium is identical with steps d;
F. produce fermentation (second order fermentation): the bacillus subtilis primary seed solution obtained by step e is inoculated in fermentation culture with 5% volume ratio, described fermentation culture is same as the first order seed liquid culture medium of step e, and condition of culture is same as step e;
the preparation of Lactobacillus delbrueckii (Lactobacillus delbrueckill):
G. make shake-flask seed: by the single colony inoculation of moral formula lactobacillus through domestication in Deshi Lactobacillus body culture medium, cultivate 36 hours, obtain shake-flask seed liquid for 37 DEG C;
Described fluid medium is the mixed liquor by following proportions: yeast extract 5 grams, peptone 5 grams, glucose 9 grams, Tween 80 0.5 gram, potassium dihydrogen phosphate 2 grams, Fructus Lycopersici esculenti juice 100ml, water 1000ml; PH=5.6,121 DEG C of sterilizings 15 minutes.
H. production primary seed solution: lactobacillus delbrueckii seed liquor step g obtained, with the inoculum concentration of 5% volume ratio, is inoculated in first order seed liquid culture medium, cultivates 40 hours in 45 DEG C, reaches 1,000,000,000/ml, obtain primary seed solution to viable count.
Described first order seed liquid culture medium prescription is the mixed liquor by following proportions: sucrose 10 grams, 25 grams, molasses, peptone 5 grams, yeast extract 5 grams, 0.5 gram, magnesium sulfate, 0.1 gram, sodium chloride, potassium dihydrogen phosphate 0.5 gram, water 1000ml;
I. produce fermentation: the seed liquor obtained by step h, with the inoculum concentration of 15% volume ratio, is inoculated in fermentation liquid culture medium, cultivate 48 hours in 40 DEG C, when dropping to 1 gram/ml with residual sugar, fermentation ends;
the preparation of Chinese medicine extraction liquid:
Described extracts of Chinese herbal medicine, be prepared from by the following step: Chinese medicine Herba polygoni hydropiperis, the Radix Euphorbiae Fischerianae (Radix Euphorbiae Ebracteolatae), Radix Clematidis, northern Cortex Dictamni, Herba Xanthii are mixed according to weight ratio 5:3:1:1:1, add the alcohol-pickled 24-72 hour of 4 ° of 10 times of volumes, again the medicine after immersion is decocted 20-30 minute under boiling water condition, it is stand-by that drop goes out medicinal liquid.
the preparation (mixed culture fermentation) of environment-friendly agent:
By weight, swamp Rhodopseudomonas: bacillus subtilis: join in mixed bacteria liquid culture medium after lactobacillus delbrueckii=3:4:7 mixing and ferment 15 days, viable count reaches 1,000,000,000/ml, during PH<4; Add the Chinese medicine extraction liquid of 1.5% volume ratio again, can complete.
Microorganism described in the present invention is buied by China General Microbiological culture presevation administrative center (CGMCC), wherein, swamp Rhodopseudomonas (Rhodo pseudmonas palustris): bacillus subtilis (Rhodo pseudmonas palustris): be numbered 1.2352, Lactobacillus delbrueckii (Lactobacillus delbrueckill): numbering 1.1482.
This environment-friendly agent is used for Sichuan Province's Yibin City destructor plant, through Yibin City environmental monitoring station, monitoring hydrogen sulfide reduces 50%, and ammonia reduces 81%.Fly significantly reduces (see figure 1).
Soot uses environment-friendly agent, and within three days, just see effect, solve fly and foul smell two hang-up, fly and foul smell obviously reduce, and create a good working environment to soot employee, periphery peasant be satisfied with.
Claims (1)
1. microbial environment protectant, is characterized in that: comprise swamp Rhodopseudomonas, bacillus subtilis, lactobacillus delbrueckii formed mixed vaccine and Chinese medicine extraction liquid; By weight, swamp Rhodopseudomonas: bacillus subtilis: lactobacillus delbrueckii=3:4:7; By volume, the ratio of Chinese medicine extraction liquid in mixed vaccine is 1%-2%;
Described swamp Rhodopseudomonas, is prepared from by the following step:
A. make shake-flask seed liquid: by the single bacterium colony of the swamp Rhodopseudomonas through domestication, be inoculated in fluid medium A, cultivate 48 hours in 30 DEG C, obtain shake-flask seed liquid;
Described fluid medium A is the mixed liquor by following proportions: sodium acetate 0.2 gram, 0.1 gram, sodium chloride, 0.1 gram, magnesium sulfate, potassium dihydrogen phosphate 0.05 gram, ammonium sulphate 0.1 gram, 0.01 gram, calcium chloride, 100ml water, and pH is 6.6; 121 DEG C of sterilizings 30 minutes, cool for subsequent use;
B. production primary seed solution: the swamp Rhodopseudomonas shake-flask seed liquid obtained by step a is with the inoculum concentration of 10% volume ratio, be inoculated into swamp Rhodopseudomonas first order seed liquid culture medium to cultivate 72 hours anaerobic environment 30 DEG C, intensity of illumination is 1000lx, bacterium number reaches 3,000,000,000/ml, obtains swamp Rhodopseudomonas primary seed solution;
Described swamp Rhodopseudomonas first order seed liquid culture medium is the mixed liquor by following proportions: potassium dihydrogen phosphate 1.0 grams, 0.1 gram, calcium chloride, calcium bicarbonate 3 grams, sodium acetate 1 gram, 1.0 grams, sodium chloride, 0.5 gram, magnesium chloride, ammonium chloride 1.0 grams, trace element storage liquid 1ml, growth cofactor storage liquid 1ml, water 1000ml; PH=6.6,121 DEG C of sterilizings 30 minutes, cool for subsequent use;
Described trace element storage liquid is: ferrous chloride 1800 milligrams, cobaltous chloride 250 milligrams, Nickel dichloride. 10 milligrams, copper chloride 10 milligrams, manganese chloride 70 milligrams, zinc chloride 100 milligrams, boric acid 500 milligrams, potassium manganate 30 milligrams, sodium selenate 10 milligrams, water 1000ml;
Described growth cofactor storage liquid is: biotin 10 milligrams, nicotinic acid 35 milligrams, thiamine 30 milligrams, para-amino benzoic acid 20 milligrams, 10 milligrams, the many aldehyde of salt acid ratio, calcium pantothenate 10 milligrams, vitamin B
12milligram, water 100ml;
C. fermentation is produced: the swamp Rhodopseudomonas primary seed solution obtained by step b is inoculated in fermentation culture D with 10% volume ratio, described fermentation culture D is same as the first order seed liquid culture medium of step b, condition of culture is same as step b, ferment 96 hours, detect when viable count has reached 3,000,000,000/ml and stop fermentation;
Finally conventional panel count detection is carried out to the viable bacteria content of swamp Rhodopseudomonas fermentation liquid;
Described bacillus subtilis, is prepared from by the following step:
D. make shake-flask seed liquid: by the single bacterium colony of the bacillus subtilis through domestication, be inoculated in fluid medium B, cultivate 48 hours in 30 DEG C, obtain shake-flask seed liquid;
Described fluid medium B is the mixed liquor by following proportions: peptone 1.0 grams, glucose 1.0 grams, Carnis Bovis seu Bubali cream 1.0 grams, potassium dihydrogen sulfate 0.5 gram, manganese sulfate 0.1 gram, water 1000ml; PH=7.5;
E. production primary seed solution: bacillus subtilis shake-flask seed liquid steps d obtained is with the inoculum concentration of 3% volume ratio, be inoculated into bacillus subtilis first order seed liquid culture medium to cultivate 20 hours anaerobic environment 30 DEG C, intensity of illumination is 1000lx, bacterium number reaches 3,000,000,000/ml, obtains bacillus subtilis primary seed solution; Bacillus subtilis first order seed liquid culture medium is identical with steps d;
F. produce fermentation: the bacillus subtilis primary seed solution obtained by step e is inoculated in fermentation culture E with 5% volume ratio, described fermentation culture E is same as the first order seed liquid culture medium of step e, and condition of culture is same as step e;
Described lactobacillus delbrueckii, is prepared from by the following step:
G. make shake-flask seed: by the single colony inoculation of lactobacillus delbrueckii through domestication in lactobacillus delbrueckii fluid medium C, cultivate 36 hours, obtain shake-flask seed liquid for 37 DEG C;
Described fluid medium C is the mixed liquor by following proportions: yeast extract 5 grams, peptone 5 grams, glucose 9 grams, Tween 80 0.5 gram, potassium dihydrogen phosphate 2 grams, Fructus Lycopersici esculenti juice 100ml, water 1000ml; PH=5.6,121 DEG C of sterilizings 15 minutes;
?h. production primary seed solution: lactobacillus delbrueckii seed liquor step g obtained, with the inoculum concentration of 5% volume ratio, is inoculated in first order seed liquid culture medium, cultivates 40 hours in 45 DEG C, reaches 1,000,000,000/ml, obtain primary seed solution to viable count;
Described first order seed liquid culture medium prescription is the mixed liquor by following proportions: sucrose 10 grams, 25 grams, molasses, peptone 5 grams, yeast extract 5 grams, 0.5 gram, magnesium sulfate, 0.1 gram, sodium chloride, potassium dihydrogen phosphate 0.5 gram, water 1000ml;
I. produce fermentation: the seed liquor obtained by step h, with the inoculum concentration of 15% volume ratio, is inoculated in fermentation liquid culture medium, cultivate 48 hours in 40 DEG C, when dropping to 1 gram/L with residual sugar, fermentation ends;
Described fermentation liquid culture medium prescription is the mixed liquor by following proportions, 40 grams, molasses, sucrose 20 grams, wheat root 18 grams, peptone 5 grams, yeast extract 5 grams, 0.5 gram, magnesium sulfate, 0.1 gram, sodium chloride, potassium dihydrogen phosphate 0.5 gram, water 1000ml;
Described extracts of Chinese herbal medicine, be prepared from by the following step: Chinese medicine Herba polygoni hydropiperis, the Radix Euphorbiae Fischerianae (Radix Euphorbiae Ebracteolatae), Radix Clematidis, northern Cortex Dictamni, Herba Xanthii are mixed according to weight ratio 5:3:1:1:1, add the alcohol-pickled 24-72 hour of 4 ° of 10 times of volumes, again the medicine after immersion is decocted 20-30 minute under boiling water condition, it is stand-by that drop goes out medicinal liquid;
Described mixed vaccine, is prepared from by the following step:
By weight, swamp Rhodopseudomonas: bacillus subtilis: join in mixed bacteria liquid culture medium after lactobacillus delbrueckii=3:4:7 mixing and ferment 15 days, viable count reaches 1,000,000,000/ml, and during pH<4 prepared by mixed liquor;
Described multi strain co cultivation base is the mixed liquor by following proportions: molasses 30%, magnesium sulfate 1%, and ferrous sulfate 0.4%, potassium dihydrogen phosphate 1%, boils and put to room temperature.
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| CN101073674A (en) * | 2007-06-22 | 2007-11-21 | 俞祖勋 | Production of deodoriferant |
| CN101948757A (en) * | 2010-09-30 | 2011-01-19 | 大连三科生物工程有限公司 | Composite microbial additive for aquatic products, preparation method thereof and use thereof |
| KR101029346B1 (en) * | 2009-10-28 | 2011-04-13 | 주식회사 엠투원 | Combined microorganism fermented liquors having the effect of reducing odor gas and antibacterial effect, and methods of using the same |
| CN102258796A (en) * | 2011-06-28 | 2011-11-30 | 黄小娃 | Microbial deodorant and preparation method thereof |
-
2012
- 2012-08-27 CN CN201210307003.0A patent/CN102813949B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101073674A (en) * | 2007-06-22 | 2007-11-21 | 俞祖勋 | Production of deodoriferant |
| KR101029346B1 (en) * | 2009-10-28 | 2011-04-13 | 주식회사 엠투원 | Combined microorganism fermented liquors having the effect of reducing odor gas and antibacterial effect, and methods of using the same |
| CN101948757A (en) * | 2010-09-30 | 2011-01-19 | 大连三科生物工程有限公司 | Composite microbial additive for aquatic products, preparation method thereof and use thereof |
| CN102258796A (en) * | 2011-06-28 | 2011-11-30 | 黄小娃 | Microbial deodorant and preparation method thereof |
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