CN113980840A - Microbial deodorant liquid for public environment and preparation method thereof - Google Patents
Microbial deodorant liquid for public environment and preparation method thereof Download PDFInfo
- Publication number
- CN113980840A CN113980840A CN202111204358.2A CN202111204358A CN113980840A CN 113980840 A CN113980840 A CN 113980840A CN 202111204358 A CN202111204358 A CN 202111204358A CN 113980840 A CN113980840 A CN 113980840A
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- mixture
- powder
- aspergillus niger
- culture medium
- spores
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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Abstract
The invention belongs to the field of microbial deodorization, and discloses a microbial deodorization liquid for public environment and a preparation method thereof.
Description
Technical Field
The invention belongs to the field of microbial deodorization, and particularly relates to a microbial deodorization liquid for public environment and a preparation method thereof.
Background
The odor of public places mainly comes from anaerobic fermentation of carbohydrates and nitrogen-sulfur-containing organic matters, and the two substances can decompose and release irritant gases with sour taste, smelly eggs, fishy smell, rotten cabbage taste and the like under the anaerobic environmental condition. The malodorous gas is generated as a result of metabolism of carbohydrates and nitrogen and sulfur containing organic matters as nutrients by a plurality of anaerobic bacteria or facultative anaerobic bacteria, and the main bacteria in the excrement are streptococcus, eubacterium and clostridium. According to the determination, the odor of the excrement consists of 168 odor compounds, for example, main odor gases in livestock and poultry farms are hydrogen sulfide, ammonia, triammonium, butanediamine, pentanediamine, methyl mercaptan, carbon disulfide, indoles, volatile fatty acids and the like, and the gases exist in the breeding farms for a long time, so that the production performance of animals is seriously influenced, and the incidence rate of respiratory tract related diseases of the livestock and poultry is greatly improved. And because the odor pollution belongs to open type unorganized emission, the components are complex, the production amount is large, the influence range is wide, the duration is long, and the gas collection is difficult, the treatment is very difficult, and the environmental pollution around the farm is greatly caused. The treatment methods for malodorous gases can be roughly classified into three categories: physical methods (masking, absorption, adsorption), chemical methods (combustion, oxidation, neutralization, washing), and biological methods. The biological deodorization method has the characteristics of high treatment efficiency, no secondary pollution, simple required equipment, convenient operation, low cost and convenient management and maintenance, and becomes an enthusiastic research direction in many countries in recent years.
The related research of the microorganisms shows that the action of the singly added microorganisms in accelerating the decomposition process of the malodorous substances is not superior to the combined action of the compound microbial flora no matter how high the activity of the singly added microorganisms is.
Disclosure of Invention
The basic principle of using the microbial deodorizing liquid to treat odor is that malodorous substances dissolved in water are absorbed into the bodies of microbes by the microbes, and the malodorous substances are degraded by the metabolic activity of the microbes.
The above purpose of the invention is realized by the following technical scheme:
a microbial deodorant liquid for public environments comprises the following components in percentage by mass:
the balance of photosynthetic bacteria liquid, and the sum of the total mass percentage is 100%.
The further preferable components in percentage by mass are as follows:
the balance of photosynthetic bacteria liquid, and the sum of the total mass percentage is 100%.
A preparation method of a microbial deodorant liquid for public environments sequentially comprises the following steps:
(1) respectively preparing bacillus subtilis powder, bacillus licheniformis powder, saccharomyces cerevisiae powder, lactobacillus powder, trichoderma viride powder, aspergillus niger powder and photosynthetic bacteria liquid;
(2) respectively weighing bacillus subtilis powder, bacillus licheniformis powder, saccharomyces cerevisiae powder, lactobacillus powder, trichoderma viride powder, aspergillus niger powder and photosynthetic bacteria liquid according to the mass percentage;
(3) adding the bacillus subtilis powder and the bacillus licheniformis powder weighed in the step (2) into the photosynthetic bacteria liquid, and stirring while adding to obtain a mixture A;
(4) sequentially adding the saccharomyces cerevisiae powder and the lactobacillus powder weighed in the step (2) into the mixture A prepared in the step (3), stirring and adding the mixture A, and fully and uniformly mixing to obtain a mixture B;
(5) slowly adding the trichoderma viride powder and the aspergillus niger powder weighed in the step (2) into the mixture B prepared in the step (4), stirring and adding the mixture B, and fully and uniformly stirring to obtain a mixture C;
(6) filtering with a 60-mesh nylon mesh, removing impurities, detecting the technical indexes of the product, and subpackaging to obtain the microbial deodorant liquid.
Further, the preparation method of the bacillus subtilis powder in the step (1) specifically comprises the following steps:
the bacillus subtilis is separated by adopting a dilution coating plate method, livestock and poultry farm colony house padding and wheat straws are respectively collected and mixed according to the mass ratio of 2:1, sterile water is added for dilution, the sterile water dosage is not required, solid substances are just submerged, the mixture is uniformly mixed and then stands, 100 mu L of supernatant is absorbed and uniformly coated on a nutrient agar culture medium plate, and the nutrient agar culture medium comprises the following components: 10g/L of peptone, 3g/L of beef extract, 5g/L of sodium chloride, 20g/L of agar, and placing the sterilized beef extract in an incubator at 30 ℃ for culturing for 48h, and observing the colony morphology, wherein the pH value is 7.2-7.4 before sterilization;
the purification of the bacillus subtilis adopts a plate marking method, single bacterial colonies which are round in colony morphology, dry in surface, and have wrinkles, milky white color and irregular edges are picked, and continuous marking is carried out on a new culture medium plate to obtain a plurality of single bacterial colonies with single morphology;
selecting the purified bacillus subtilis, and carrying out microscopic morphological identification and biochemical characteristic analysis;
after identification, transferring the purified bacillus subtilis into a nutrient agar test tube inclined plane, culturing for 48-72 hours at 30 ℃ until the whole inclined plane is full of lawn, and continuously performing propagation by using the method, wherein the method is first-class seed culture;
selecting a first-class strain which grows mature, inoculating the first-class strain into a 2000mL conical flask containing 500mL of amplification culture solution under an aseptic condition, wherein the amplification culture solution comprises the following components: 10g/L of glucose, 10g/L of bran, 5g/L of peptone, 2g/L of beef extract and Na2HPO42.5g/L，KH2PO41g/L, Tween-801.5g/L, pH 7.2-7.4 before sterilization, shaking culture at 28 deg.C for 48h, sampling and counting, the effective viable count in bacterial liquid is not less than 5 × 1010Per gram;
the three-stage solid fermentation of the bacillus subtilis adopts a deep ventilation fermentation method, and the sterilized and cooled solid fermentation material and the secondary bacterial liquid are uniformly mixed according to the mass ratio of 1:10, wherein the solid fermentation material comprises the following components: 6kg of bran, 1kg of corn flour, 1kg of rice hull, 1.5kg of molasses, 0.5kg of soybean peptone or fish peptone and KH2PO41g，MgSO40.5g, the water content is 35-40%, the pH value before sterilization is 7.2-7.4, the mixture is flatly paved in a ventilation fermentation tank, the material depth is 15-20 cm, the temperature is 28 ℃ for 48 hours, and when the material core temperature is more than or equal to 45 ℃, the material turning operation is carried out once;
after fermentation, drying, crushing, sieving with 60 mesh sieve to obtain final product with effective viable count not less than 5 × 1010The water content is 8-9% per gram.
Further, the preparation method of the bacillus licheniformis powder in the step (1) specifically comprises the following steps:
separating the bacillus licheniformis by adopting a dilution coating plate method, respectively collecting soil of a colony house of a farm and culture wastewater, mixing according to the mass ratio of 3:1, adding sterile water for dilution, wherein the sterile water does not have the requirement on the using amount, immersing solid matters, uniformly mixing, standing, absorbing 100 mu L of supernatant, uniformly coating the supernatant on a nutrient agar medium plate, placing the nutrient agar medium plate in an incubator at 30 ℃ for culturing for 48 hours, and observing the colony morphology;
the purification of the bacillus licheniformis adopts a plate scribing method, single bacterial colonies with round colony morphology, dry surface, roughness and irregular edge are picked, and continuous scribing is carried out on a new culture medium plate to obtain a plurality of single bacterial colonies with single morphology;
selecting the purified bacillus licheniformis, and carrying out microscopic morphological identification and biochemical characteristic analysis;
after identification, transferring the purified bacillus licheniformis into a nutrient agar test tube inclined plane, culturing for 48-72 h at 30 ℃ until the whole inclined plane is full of lawn, and continuously performing propagation by using the method, wherein the step is first-class culture;
selecting mature first-class strain, inoculating to 2000mL conical flask containing 500mL amplification culture solution under aseptic condition, shake-culturing at 28 deg.C for 48 hr, sampling, and counting to obtain bacterial solution with effective viable count of 5 × 10 or more10Per gram;
the three-stage solid fermentation of the bacillus licheniformis adopts a deep ventilation fermentation method, the sterilized and cooled solid fermentation material and the secondary bacterial liquid are uniformly mixed according to the mass ratio of 1:10, and the solid fermentation material comprises the following components: 6kg of bran, 1kg of corn flour, 1kg of rice hull, 1.5kg of molasses, 0.5kg of soybean peptone or fish peptone and KH2PO41g，MgSO40.5g, the water content is 35-40%, the pH value before sterilization is 7.2-7.4, the mixture is flatly paved in a ventilation fermentation tank, the material depth is 15-20 cm, the fermentation is carried out for 48 hours at 28 ℃, and when the material core temperature is more than or equal to 45 ℃, the material turning operation is carried out once;
after fermentation, drying, crushing, sieving with 60 mesh sieve to obtain final product with effective viable count not less than 5 × 1010Per gram, the water content is 8-9%;
further, the preparation method of the saccharomyces cerevisiae powder in the step (1) comprises the following specific steps:
the saccharomyces cerevisiae is saccharomyces cerevisiae CICCNo.1015, has the characteristic of fermenting galactose, raffinose and other small molecular saccharides, and is transferred into a test tube inclined plane of a YPD culture medium, wherein the YPD culture medium comprises the following components: 10g/L of yeast extract, 20g/L of peptone, 20g/L of glucose and 20g/L of agar powder, wherein the pH value before sterilization is 6.5-6.8, and the mixture is cultured at 30 ℃ for 48-72 hours until the whole inclined plane is full of bacterial lawn;
selecting mature first-class strain, inoculating into 2000mL conical flask containing 500mLYPD culture solution under aseptic condition, shake-culturing at 28 deg.C for 48 hr, sampling, and counting to obtain bacterial solution with effective viable count of 2 × 10 or more10Per gram;
the three-stage solid fermentation of the saccharomyces cerevisiae adopts a deep ventilation fermentation method, the sterilized and cooled solid fermentation material and the secondary bacterial liquid are uniformly mixed according to the mass ratio of 1:10, and the solid fermentation material comprises the following components: 5kg of bran, 1kg of wheat flour, 2kg of glucose, 1kg of malt flour, 1kg of soybean peptone or fish peptone and KH2PO41g，MgSO40.5g, the water content is 35-40%, the pH value before sterilization is 6.5-6.8, the mixture is flatly paved in a ventilation fermentation tank, the depth of the material is 15-20 cm, the fermentation is carried out for 72 hours at the temperature of 28 ℃, and when the temperature of a material core is more than or equal to 40 ℃, the material turning operation is carried out once;
after fermentation, drying, crushing, sieving with 60 mesh sieve to obtain product with effective viable count not less than 2 × 1010Per gram, the water content is 8-9%;
preferably, the lactobacillus powder is a commercial agricultural grade lactobacillus solid fermentation product, and the number of effective viable bacteria in the product is more than or equal to 5 multiplied by 1010Per gram, the water content is 8-9%;
further, the preparation method of the trichoderma viride powder in the step (1) specifically comprises the following steps:
separating trichoderma viride by adopting a dilution coating flat plate method, respectively collecting bedding materials and rotten forage grass of a farm colony, uniformly mixing according to a mass ratio of 1:1, weighing 10g of the mixture, adding the mixture into 90mL of sterile water, placing the mixture on a shaking table with a rotation number of 180r/min for shaking for 1h, standing, absorbing 100 mu L of supernatant, uniformly coating the supernatant on a trichoderma screening culture medium flat plate, and preparing the trichoderma screening culture medium according to the following method: adding 100g of fresh potato blocks into 1l of pure water, decocting for 10-15 min, cooling, homogenizing, adding pure water to 1000ml, sequentially adding 20g of brown sugar and KH2PO41.2g，MgSO40.6g, 0.008g of thiamine, 0.1g of chloramphenicol, 20g of agar,placing the sterilized mixture in an incubator at 28 ℃ for culturing for 72h under the condition that the pH value is 6.5-6.8 before sterilization, and observing the colony morphology;
the trichoderma viride is purified by adopting a single spore colony method, namely when trichoderma colonies grow on a plate and generate green spores, gently picking the spores by using an inoculation shovel, uniformly inoculating the spores on a trichoderma enhanced culture medium plate, wherein the trichoderma enhanced culture medium is prepared by the following method: adding 100g of fresh potato blocks into every 1L of soil leaching solution, decocting for 10-15 min, cooling, homogenizing, supplementing pure water to 1000mL, sequentially adding 20g of brown sugar and KH2PO41.2g，MgSO40.6g, 0.008g of thiamine, 0.1g of chloramphenicol and 20g of agar, wherein the pH value before sterilization is 6.5-6.8, the mixture is placed in an incubator at 28 ℃ for culturing for 72 hours, and the mixture is purified for multiple times by the method so as to obtain a plurality of single colonies with single morphology;
selecting the trichoderma spores obtained through purification, and carrying out microscopic morphological identification;
after identification, transferring the purified trichoderma viride spores into a test tube inclined plane of a trichoderma enhanced culture medium, culturing for 72 hours at 28 ℃ until hyphae grow on the whole inclined plane and mature spores are generated, and continuously performing propagation by using the method, namely primary strain culture;
selecting mature first-class seeds, scraping spores on the inclined plane of the test tube by using an inoculating shovel under aseptic condition, and inoculating the spores into a 500mL conical flask filled with a mold amplification culture medium, wherein the mold amplification culture medium comprises the following components: 32.5g of bran, 25g of corn flour, 7.5g of rice hull, 5g of soybean peptone or fish peptone, 42-47% of water content, 6.5-6.8 of pH before sterilization, culturing for 72h at 28 ℃ until culture medium materials are full of trichoderma viride conidia, sampling and counting, wherein the number of the trichoderma viride conidia is more than or equal to 5 multiplied by 109Per gram;
the three-stage solid fermentation of the trichoderma viride adopts a deep ventilation fermentation method, and the sterilized and cooled solid fermentation material and the secondary strain are uniformly mixed according to the mass ratio of 1:8, wherein the solid fermentation material comprises the following components: 3.25kg of bran, 2.5kg of corn flour, 0.75kg of rice hull, 0.5kg of soybean peptone or fish peptone and KH2PO41g，MgSO40.5g, the water content is 42-47%, the pH value before sterilization is 6.5-6.8, the mixture is flatly paved in a ventilation fermentation tank, the material depth is 15-20 cm, and the temperature is 28 DEG CFermenting for 72h, and turning once when the temperature of the material core is more than or equal to 40 ℃;
after fermentation, drying, crushing, sieving with a 60-mesh sieve, wherein the number of conidia of the Trichoderma viride in the product is more than or equal to 5 multiplied by 109Per gram, the water content is 8-9%;
further, the preparation method of the aspergillus niger bacterial powder in the step (1) specifically comprises the following steps:
aspergillus niger separation adopts a dilution coating flat plate method, surface soil of an organic fertilizer decomposition field is respectively collected, 10g of the organic fertilizer decomposition field is weighed and added into 90mL of sterile water, the sterile water is placed on a shaking table with the rotation number of 180r/min for shaking for 1h, standing is carried out, 100 mu L of supernate is absorbed and is uniformly coated on an Aspergillus niger selective culture medium flat plate, and the Aspergillus niger selective culture medium comprises the following components: (NH)4)2SO40.5g/L, 150g/L brown sugar and KH2PO41g/L，MgSO40.5g/L, 0.1g/L chloramphenicol, 30g/L agar, pH 6.5-6.8 before sterilization, culturing in an incubator at 28 deg.C for 72h, and observing colony morphology;
the aspergillus niger is purified by adopting a monospore colony method, namely when an aspergillus niger colony grows on a flat plate and generates black spores, the spores are lightly picked by using an inoculation shovel, evenly inoculated on the aspergillus niger selective culture medium flat plate and cultured in an incubator at 28 ℃ for 72 hours, and the purification is carried out for a plurality of times according to the method so as to obtain a plurality of single colonies with single shape;
selecting the purified aspergillus niger spores, and carrying out microscopic morphological identification;
after identification, transferring the purified aspergillus niger spores into an aspergillus niger selective culture medium test tube inclined plane, culturing for 72 hours at 28 ℃ until the whole inclined plane is full of hyphae and mature conidia are generated, and continuously performing propagation by using the method, namely primary strain culture;
selecting mature first-class seeds, scraping spores on the inclined plane of the test tube by using an inoculating shovel under aseptic condition, and inoculating the spores into a 500mL conical flask filled with a mold amplification culture medium, wherein the mold amplification culture medium comprises the following components: 32.5g of bran, 25g of corn flour, 7.5g of rice hull, 5g of soybean peptone or fish peptone, 42-47% of water content, 6.5-6.8 of pH before sterilization, culturing for 72h at 28 ℃ until the culture medium materials are full of Aspergillus niger conidia, sampling and counting,the number of Aspergillus niger conidium is not less than 5 × 109Per gram;
the three-stage solid fermentation of the aspergillus niger adopts a deep ventilation fermentation method, the sterilized and cooled solid fermentation material and the secondary strain are uniformly mixed according to the mass ratio of 1:8, and the solid fermentation material comprises the following components: 3.25kg of bran, 2.5kg of corn flour, 0.75kg of rice hull, 0.5kg of soybean peptone or fish peptone and KH2PO41g，MgSO40.5g, the water content is 42-47%, the pH value before sterilization is 6.5-6.8, the mixture is flatly paved in a ventilation fermentation tank, the material depth is 15-20 cm, the fermentation is carried out for 72 hours at 28 ℃, and when the material core temperature is more than or equal to 40 ℃, the material turning operation is carried out once;
after fermentation, drying, crushing and sieving with a 60-mesh sieve, wherein the number of aspergillus niger conidia in the product is more than or equal to 5 multiplied by 109Per gram, the water content is 8-9%;
further, the preparation method of the photosynthetic bacteria liquid in the step (1) specifically comprises the following steps:
(1) separately weighing NaHCO31g, sodium acetate 1g, NaCl1g, NH4Cl1g、MgSO40.3g、K2HPO40.2g, 0.2g of sodium propionate, 0.25g of malic acid, 0.2g of peptone, 0.1g of yeast extract and 1000mL of deionized water;
(2) adding the yeast extract and peptone weighed in the step (1) into deionized water, heating, and uniformly stirring to obtain a mixture D;
(3) weighing NaHCO in the step (1)3Sodium acetate, NaCl, NH4Cl、MgSO4、K2HPO4Sequentially adding sodium propionate and malic acid into the mixture D prepared in the step (2), and fully and uniformly mixing to obtain a mixture E;
(4) respectively weighing 10g of gamma-aminobutyric acid and 0.2g of glutamic acid, and dissolving in 1L of deionized water to obtain a mixture F;
(5) adding 1mL of mixture F into each liter of mixture E, adding the mixture F prepared in the step (4) into the mixture E prepared in the step (3), and fully and uniformly stirring to obtain a mixture G;
(6) collecting water samples on the surface layer of the breeding wastewater or the septic tank, and according to the water samples: the mixture G is in a volume ratio of 1: 6-1: 8, fully mixed, subpackaged in a transparent container, and subjected to light anaerobic culture;
further, the light anaerobic culture conditions of the step (6) are as follows: stirring or shaking regularly at 28-32 ℃ and 4000Lux, and culturing for 7-10 days.
Compared with the prior art, the invention has the beneficial effects that:
various microorganisms in the deodorant liquid play an important role, and a symbiotic relation of stable state and complete functions is formed.
The photosynthetic bacteria are anaerobic autotrophic microorganisms, mainly comprise rhodospirillum, chromobacteria, green bacillus, green filamentous bacteria and the like, and the metabolic types of each type of bacteria are different, so that the photosynthetic bacteria have multiple purposes. In the aspect of environmental protection and deodorization, the photosynthetic bacteria can utilize peculiar smell substances such as hydrogen sulfide and the like as a reducing agent to synthesize carbon dioxide into organic matters through a photosynthetic phosphorylation process; they can also synthesize proteins, nucleic acids, and the like using ammonia or the like as a nitrogen source, thereby converting malodorous substances.
The bacillus microorganisms are also a kind of common probiotics and are widely applied to the industries of planting, breeding, food fermentation, medicine and health, environmental management and the like.
The lactobacillus and yeast microorganisms can respectively use diversified metabolic pathways to synthesize micromolecular organic matters such as methionine, cysteine, trimethylamine, dithiol and the like, or self components such as polypeptide, protein and the like, or decompose the micromolecular organic matters into inorganic matters for other microorganisms such as photosynthetic bacteria and the like under anaerobic and aerobic conditions. In addition, they can also produce metabolites such as organic acid, alcohol and the like, inhibit the growth of harmful bacteria and reduce the generation of harmful gases.
The utilization of mould microorganisms for environmental management is a new research direction in the field of environmental protection in recent years, the mould metabolism types are extremely diverse, and the produced high-activity enzymes are often secreted outside cells to produce effects, so that the mould has the advantages of wide application range, high metabolic efficiency, rapid growth and the like. Decomposing animal and plant residues, excrement, kitchen waste, micro plastic and the like in the sewage by using aspergillus niger; trichoderma is widely used for soil improvement and sewage treatment, and in the growth process, the Trichoderma can not only secrete hydrolase such as protease, peptidase, cellulase, hemicellulase, chitinase and the like, but also can generate various secondary metabolites such as phenethyl alcohol, pyrone, biostimulan and the like, and can effectively promote the growth of other probiotics while decomposing harmful pollutants. The existing microbial deodorant liquid is not found to contain a mould component, and the mould is added into photosynthetic bacteria, bacillus, lactobacillus and saccharomycetes, so that various microorganisms have synergistic action, and a microecological flora for efficiently degrading harmful pollutants is established. The deodorizing composite bacterial liquid can inhibit putrefying bacteria and pathogenic bacteria, eliminate odor substances and form a good environment, thereby improving the production level and the life quality of the whole society and protecting the global environment and beautiful home of human beings.
Drawings
FIG. 1 is a graph showing the effect of spraying the product for one week on the removal of hydrogen sulfide (a) and ammonia (b) in mg/m3；
FIG. 2 is a graph showing the results of an experimental verification of the accelerated decomposition effect according to the present invention;
FIG. 3 is a graph of the results of the manure germination experiment using the product to treat a non-thoroughly decomposed manure.
Detailed Description
The invention is described in more detail below with reference to specific examples, without limiting the scope of the invention. Unless otherwise specified, the experimental methods adopted by the invention are all conventional methods, and experimental equipment, materials, reagents and the like used in the experimental method can be obtained from commercial sources.
EXAMPLE 1 preparation of microbial deodorant solution of the present invention
(1) Respectively preparing bacillus subtilis powder, bacillus licheniformis powder, saccharomyces cerevisiae powder, lactobacillus powder, trichoderma viride powder, aspergillus niger powder and photosynthetic bacteria liquid;
the preparation method of the bacillus subtilis powder comprises the following steps:
the bacillus subtilis is separated by adopting a dilution coating plate method, livestock and poultry farm colony house padding and wheat straws are respectively collected and mixed according to the mass ratio of 2:1, sterile water is added for dilution, the sterile water dosage is not required, solid substances are just submerged, the mixture is uniformly mixed and then stands, 100 mu L of supernatant is absorbed and uniformly coated on a nutrient agar culture medium plate, and the nutrient agar culture medium comprises the following components: 10g/L of peptone, 3g/L of beef extract, 5g/L of sodium chloride, 20g/L of agar, and placing the sterilized beef extract in an incubator at 30 ℃ for culturing for 48h, and observing the colony morphology, wherein the pH value is 7.2-7.4 before sterilization;
the purification of the bacillus subtilis adopts a plate marking method, single bacterial colonies which are round in colony morphology, dry in surface, and have wrinkles, milky white color and irregular edges are picked, and continuous marking is carried out on a new culture medium plate to obtain a plurality of single bacterial colonies with single morphology;
selecting the purified bacillus subtilis, and carrying out microscopic morphological identification and biochemical characteristic analysis;
after identification, transferring the purified bacillus subtilis into a nutrient agar test tube inclined plane, culturing for 48-72 hours at 30 ℃ until the whole inclined plane is full of lawn, and continuously performing propagation by using the method, wherein the method is first-class seed culture;
selecting a first-class strain which grows mature, inoculating the first-class strain into a 2000mL conical flask containing 500mL of amplification culture solution under an aseptic condition, wherein the amplification culture solution comprises the following components: 10g/L of glucose, 10g/L of bran, 5g/L of peptone, 2g/L of beef extract and Na2HPO42.5g/L，KH2PO41g/L, Tween-801.5g/L, pH 7.2-7.4 before sterilization, shaking culture at 28 deg.C for 48h, sampling and counting, the effective viable count in bacterial liquid is not less than 5 × 1010Per gram;
the three-stage solid fermentation of the bacillus subtilis adopts a deep ventilation fermentation method, and the sterilized and cooled solid fermentation material and the secondary bacterial liquid are uniformly mixed according to the mass ratio of 1:10, wherein the solid fermentation material comprises the following components: 6kg of bran, 1kg of corn flour, 1kg of rice hull, 1.5kg of molasses, 0.5kg of soybean peptone or fish peptone and KH2PO41g，MgSO40.5g, the water content is 35-40%, the pH value before sterilization is 7.2-7.4, and the mixture is spread in a ventilation fermentation wayIn the pool, the depth of the material is 15-20 cm, the material is heated at 28 ℃ for 48 hours, and when the temperature of a material core is more than or equal to 45 ℃, the material turning operation is carried out once;
after fermentation, drying, crushing, sieving with 60 mesh sieve to obtain final product with effective viable count not less than 5 × 1010The water content is 8-9% per gram.
The preparation method of the bacillus licheniformis powder comprises the following steps:
separating the bacillus licheniformis by adopting a dilution coating plate method, respectively collecting soil of a colony house of a farm and culture wastewater, mixing according to the mass ratio of 3:1, adding sterile water for dilution, wherein the sterile water does not have the requirement on the using amount, immersing solid matters, uniformly mixing, standing, absorbing 100 mu L of supernatant, uniformly coating the supernatant on a nutrient agar medium plate, placing the nutrient agar medium plate in an incubator at 30 ℃ for culturing for 48 hours, and observing the colony morphology;
the purification of the bacillus licheniformis adopts a plate scribing method, single bacterial colonies with round colony morphology, dry surface, roughness and irregular edge are picked, and continuous scribing is carried out on a new culture medium plate to obtain a plurality of single bacterial colonies with single morphology;
selecting the purified bacillus licheniformis, and carrying out microscopic morphological identification and biochemical characteristic analysis;
after identification, transferring the purified bacillus licheniformis into a nutrient agar test tube inclined plane, culturing for 48-72 h at 30 ℃ until the whole inclined plane is full of lawn, and continuously performing propagation by using the method, wherein the step is first-class culture;
selecting mature first-class strain, inoculating to 2000mL conical flask containing 500mL amplification culture solution under aseptic condition, shake-culturing at 28 deg.C for 48 hr, sampling, and counting to obtain bacterial solution with effective viable count of 5 × 10 or more10Per gram;
the three-stage solid fermentation of the bacillus licheniformis adopts a deep ventilation fermentation method, the sterilized and cooled solid fermentation material and the secondary bacterial liquid are uniformly mixed according to the mass ratio of 1:10, and the solid fermentation material comprises the following components: 6kg of bran, 1kg of corn flour, 1kg of rice hull, 1.5kg of molasses, 0.5kg of soybean peptone or fish peptone and KH2PO41g，MgSO40.5g, water content 35-40%, pH 7.2-7.4 before sterilization, and spreading in ventilationIn the fermentation tank, the depth of the material is 15-20 cm, the material is fermented for 48 hours at 28 ℃, and when the temperature of a material core is more than or equal to 45 ℃, the material is turned once;
after fermentation, drying, crushing, sieving with 60 mesh sieve to obtain final product with effective viable count not less than 5 × 1010Per gram, the water content is 8-9%;
the preparation method of the saccharomyces cerevisiae bacterial powder comprises the following steps:
the saccharomyces cerevisiae is saccharomyces cerevisiae CICCNo.1015, has the characteristic of fermenting galactose, raffinose and other small molecular saccharides, and is transferred into a test tube inclined plane of a YPD culture medium, wherein the YPD culture medium comprises the following components: 10g/L of yeast extract, 20g/L of peptone, 20g/L of glucose and 20g/L of agar powder, wherein the pH value before sterilization is 6.5-6.8, and the mixture is cultured at 30 ℃ for 48-72 hours until the whole inclined plane is full of bacterial lawn;
selecting mature first-class strain, inoculating into 2000mL conical flask containing 500mLYPD culture solution under aseptic condition, shake-culturing at 28 deg.C for 48 hr, sampling, and counting to obtain bacterial solution with effective viable count of 2 × 10 or more10Per gram;
the three-stage solid fermentation of the saccharomyces cerevisiae adopts a deep ventilation fermentation method, the sterilized and cooled solid fermentation material and the secondary bacterial liquid are uniformly mixed according to the mass ratio of 1:10, and the solid fermentation material comprises the following components: 5kg of bran, 1kg of wheat flour, 2kg of glucose, 1kg of malt flour, 1kg of soybean peptone or fish peptone and KH2PO41g，MgSO40.5g, the water content is 35-40%, the pH value before sterilization is 6.5-6.8, the mixture is flatly paved in a ventilation fermentation tank, the depth of the material is 15-20 cm, the fermentation is carried out for 72 hours at the temperature of 28 ℃, and when the temperature of a material core is more than or equal to 40 ℃, the material turning operation is carried out once;
after fermentation, drying, crushing, sieving with 60 mesh sieve to obtain product with effective viable count not less than 2 × 1010Per gram, the water content is 8-9%;
preferably, the lactobacillus powder is a commercial agricultural grade lactobacillus solid fermentation product, and the number of effective viable bacteria in the product is more than or equal to 5 multiplied by 1010Per gram, the water content is 8-9%;
the preparation method of the trichoderma viride powder comprises the following specific steps:
separating trichoderma viride by adopting a dilution coating flat plate method, respectively collecting bedding materials and rotten forage grass of a farm colony, uniformly mixing according to a mass ratio of 1:1, weighing 10g of the mixture, adding the mixture into 90mL of sterile water, placing the mixture on a shaking table with a rotation number of 180r/min for shaking for 1h, standing, absorbing 100 mu L of supernatant, uniformly coating the supernatant on a trichoderma screening culture medium flat plate, and preparing the trichoderma screening culture medium according to the following method: adding 100g of fresh potato blocks into 1l of pure water, decocting for 10-15 min, cooling, homogenizing, adding pure water to 1000ml, sequentially adding 20g of brown sugar and KH2PO41.2g，MgSO40.6g, 0.008g of thiamine, 0.1g of chloramphenicol and 20g of agar, wherein the pH value before sterilization is 6.5-6.8, the mixture is placed in an incubator at 28 ℃ for culturing for 72 hours, and the colony morphology is observed;
the trichoderma viride is purified by adopting a single spore colony method, namely when trichoderma colonies grow on a plate and generate green spores, gently picking the spores by using an inoculation shovel, uniformly inoculating the spores on a trichoderma enhanced culture medium plate, wherein the trichoderma enhanced culture medium is prepared by the following method: adding 100g of fresh potato blocks into every 1L of soil leaching solution, decocting for 10-15 min, cooling, homogenizing, supplementing pure water to 1000mL, sequentially adding 20g of brown sugar and KH2PO41.2g，MgSO40.6g, 0.008g of thiamine, 0.1g of chloramphenicol and 20g of agar, wherein the pH value before sterilization is 6.5-6.8, the mixture is placed in an incubator at 28 ℃ for culturing for 72 hours, and the mixture is purified for multiple times by the method so as to obtain a plurality of single colonies with single morphology;
selecting the trichoderma spores obtained through purification, and carrying out microscopic morphological identification;
after identification, transferring the purified trichoderma viride spores into a test tube inclined plane of a trichoderma enhanced culture medium, culturing for 72 hours at 28 ℃ until hyphae grow on the whole inclined plane and mature spores are generated, and continuously performing propagation by using the method, namely primary strain culture;
selecting mature first-class seeds, scraping spores on the inclined plane of the test tube by using an inoculating shovel under aseptic condition, and inoculating the spores into a 500mL conical flask filled with a mold amplification culture medium, wherein the mold amplification culture medium comprises the following components: 32.5g of bran, 25g of corn flour, 7.5g of rice hull, 5g of soybean peptone or fish peptone, 42-47% of water content, 6.5-6.8 of pH before sterilization, and culturing for 72h at 28 ℃ until culture is completedThe culture medium material is full of trichoderma viride conidia, sampling and counting are carried out, the number of the trichoderma viride conidia is more than or equal to 5 multiplied by 109Per gram;
the three-stage solid fermentation of the trichoderma viride adopts a deep ventilation fermentation method, and the sterilized and cooled solid fermentation material and the secondary strain are uniformly mixed according to the mass ratio of 1:8, wherein the solid fermentation material comprises the following components: 3.25kg of bran, 2.5kg of corn flour, 0.75kg of rice hull, 0.5kg of soybean peptone or fish peptone and KH2PO41g，MgSO40.5g, the water content is 42-47%, the pH value before sterilization is 6.5-6.8, the mixture is flatly paved in a ventilation fermentation tank, the material depth is 15-20 cm, the fermentation is carried out for 72 hours at 28 ℃, and when the material core temperature is more than or equal to 40 ℃, the material turning operation is carried out once;
after fermentation, drying, crushing, sieving with a 60-mesh sieve, wherein the number of conidia of the Trichoderma viride in the product is more than or equal to 5 multiplied by 109Per gram, the water content is 8-9%;
the preparation method of the Aspergillus niger powder specifically comprises the following steps:
aspergillus niger separation adopts a dilution coating flat plate method, surface soil of an organic fertilizer decomposition field is respectively collected, 10g of the organic fertilizer decomposition field is weighed and added into 90mL of sterile water, the sterile water is placed on a shaking table with the rotation number of 180r/min for shaking for 1h, standing is carried out, 100 mu L of supernate is absorbed and is uniformly coated on an Aspergillus niger selective culture medium flat plate, and the Aspergillus niger selective culture medium comprises the following components: (NH)4)2SO40.5g/L, 150g/L brown sugar and KH2PO41g/L，MgSO40.5g/L, 0.1g/L chloramphenicol, 30g/L agar, pH 6.5-6.8 before sterilization, culturing in an incubator at 28 deg.C for 72h, and observing colony morphology;
the aspergillus niger is purified by adopting a monospore colony method, namely when an aspergillus niger colony grows on a flat plate and generates black spores, the spores are lightly picked by using an inoculation shovel, evenly inoculated on the aspergillus niger selective culture medium flat plate and cultured in an incubator at 28 ℃ for 72 hours, and the purification is carried out for a plurality of times according to the method so as to obtain a plurality of single colonies with single shape;
selecting the purified aspergillus niger spores, and carrying out microscopic morphological identification;
after identification, transferring the purified aspergillus niger spores into an aspergillus niger selective culture medium test tube inclined plane, culturing for 72 hours at 28 ℃ until the whole inclined plane is full of hyphae and mature conidia are generated, and continuously performing propagation by using the method, namely primary strain culture;
selecting mature first-class seeds, scraping spores on the inclined plane of the test tube by using an inoculating shovel under aseptic condition, and inoculating the spores into a 500mL conical flask filled with a mold amplification culture medium, wherein the mold amplification culture medium comprises the following components: 32.5g of bran, 25g of corn flour, 7.5g of rice hull, 5g of soybean peptone or fish peptone, 42-47% of water content, 6.5-6.8 of pH before sterilization, culturing for 72h at 28 ℃ until the culture medium is full of Aspergillus niger conidia, sampling and counting, wherein the number of Aspergillus niger conidia is more than or equal to 5 multiplied by 109Per gram;
the three-stage solid fermentation of the aspergillus niger adopts a deep ventilation fermentation method, the sterilized and cooled solid fermentation material and the secondary strain are uniformly mixed according to the mass ratio of 1:8, and the solid fermentation material comprises the following components: 3.25kg of bran, 2.5kg of corn flour, 0.75kg of rice hull, 0.5kg of soybean peptone or fish peptone and KH2PO41g，MgSO40.5g, the water content is 42-47%, the pH value before sterilization is 6.5-6.8, the mixture is flatly paved in a ventilation fermentation tank, the material depth is 15-20 cm, the fermentation is carried out for 72 hours at 28 ℃, and when the material core temperature is more than or equal to 40 ℃, the material turning operation is carried out once;
after fermentation, drying, crushing and sieving with a 60-mesh sieve, wherein the number of aspergillus niger conidia in the product is more than or equal to 5 multiplied by 109Per gram, the water content is 8-9%;
the preparation method of the photosynthetic bacteria liquid comprises the following steps:
(1) separately weighing NaHCO31g, sodium acetate 1g, NaCl1g, NH4Cl1g、MgSO40.3g、K2HPO40.2g, 0.2g of sodium propionate, 0.25g of malic acid, 0.2g of peptone, 0.1g of yeast extract and 1000mL of deionized water;
(2) adding the yeast extract and peptone weighed in the step (1) into deionized water, heating, and uniformly stirring to obtain a mixture D;
(3) weighing NaHCO in the step (1)3Sodium acetate, NaCl, NH4Cl、MgSO4、K2HPO4Sequentially adding sodium propionate and malic acid into the mixture D prepared in the step (2), and fully and uniformly mixing to obtain a mixture E;
(4) respectively weighing 10g of gamma-aminobutyric acid and 0.2g of glutamic acid, and dissolving in 1L of deionized water to obtain a mixture F;
(5) adding 1mL of mixture F into each liter of mixture E, adding the mixture F prepared in the step (4) into the mixture E prepared in the step (3), and fully and uniformly stirring to obtain a mixture G;
(6) collecting water samples on the surface layer of the breeding wastewater or the septic tank, and according to the water samples: the mixture G is in a volume ratio of 1: 6-1: 8, fully mixed, subpackaged in a transparent container, and subjected to light anaerobic culture;
further, the light anaerobic culture conditions of the step (6) are as follows: stirring or shaking regularly at 28-32 ℃ and 4000Lux, and culturing for 7-10 days.
(2) Respectively weighing bacillus subtilis powder, bacillus licheniformis powder, saccharomyces cerevisiae powder, lactobacillus powder, trichoderma viride powder, aspergillus niger powder and photosynthetic bacteria liquid according to the following mass percentage;
the balance of photosynthetic bacteria liquid, and the sum of the total mass percentage is 100%.
(3) Adding the bacillus subtilis powder and the bacillus licheniformis powder weighed in the step (2) into the photosynthetic bacteria liquid, and stirring while adding to obtain a mixture A;
(4) sequentially adding the saccharomyces cerevisiae powder and the lactobacillus powder weighed in the step (2) into the mixture A prepared in the step (3), stirring and adding the mixture A, and fully and uniformly mixing to obtain a mixture B;
(5) slowly adding the trichoderma viride powder and the aspergillus niger powder weighed in the step (2) into the mixture B prepared in the step (4), stirring and adding the mixture B, and fully and uniformly stirring to obtain a mixture C;
(6) filtering with a 60-mesh nylon mesh, removing impurities, detecting the technical indexes of the product, and subpackaging to obtain the microbial deodorant liquid.
Example 2 technical specifications of the microbial deodorant solution of the invention are as follows:
main technical indexes of the product
| Item | Index (I) | |
| Effective viable count (cfu) of photosynthetic bacteria in hundred million/mL | ≥ | 20.0 |
| Effective viable count (cfu) of Bacillus subtilis in hundred million/mL | ≥ | 5.0 |
| Effective viable count (cfu) of Bacillus licheniformis in hundred million/mL | ≥ | 5.0 |
| Effective viable count (cfu) of lactic acid bacteria in hundred million/mL | ≥ | 5.0 |
| Effective viable count (cfu) of yeast in hundred million/mL | ≥ | 2.0 |
| Number of spores of Trichoderma viride in hundred million/mL | ≥ | 0.2 |
| Aspergillus niger spore number of hundred million/mL | ≥ | 0.2 |
| The rate of mixed bacteria% | ≤ | 10 |
| pH value | 6.5-7.5 | |
| Water content% | ≤ | 30.0 |
| Fecal coliform population (number/mL) | ≤ | 100 |
| Aflatoxin B1(mg/kg) | ＜ | 0.02 |
| Death rate (%) of roundworm egg | ≥ | 95 |
| Arsenic and its compounds (As)/(mg/kg) | ≤ | 75 |
| Cadmium (and its compounds (in Cd)/(mg/kg) | ≤ | 10 |
| Lead and its compounds (in terms of Pb)/(mg/kg) | ≤ | 100 |
| Chromium and its compounds (in terms of Cr)/(mg/kg) | ≤ | 15 |
| Mercury and its compounds (in Hg)/(mg/kg) | ≤ | 5 |
| Shelf life of month | ≥ | 12 |
Embodiment 3 the odor removing effect of the product on livestock and poultry farms
The use method of the microbial deodorant liquid comprises the following steps:
the product is mainly used for producing waste gas and odor places such as livestock and poultry farms, public washrooms, septic tanks, garbage stations, rotten plants and the like; the cleaning and deodorizing work of toilets, sewage pools, family toilets, sewage pipes, ditches and the like in hotels, markets, supermarkets and the like has the following specific use conditions:
note: 1. the usage amount, times and dilution rate are common reference values.
2. The product contains multiple bioactive components, and can be used as soon as possible after being unpacked, and is not suitable for long-term storage;
3. the product can not be mixed with pesticide, copper-containing substance and disinfectant;
4. if a small amount of precipitate is normal, the product should be shaken well before use;
5. please store in dark and cool place with validity period of 12 months.
The odor removal effect of the product on livestock and poultry farms is shown in figure 1, after the product is diluted by 10 times, the spraying experiment is carried out, the foul odor of the septic tank begins to be lightened after 3 days, the foul odor is remarkably lightened after 5 days, no odor exists basically after 1 week, the number of mosquitoes and flies is reduced greatly, the flying scene cannot be seen, the detection result of the content of hydrogen sulfide in the septic tank also shows that the product has remarkable effect on removing hydrogen sulfide, and the removal effect of ammonia is also remarkable.
Example 4 Effect of the product on decomposition of feces and urine
The manure is decomposed by the conventional technology, and the decomposed materials produced have toxic action on the seedlings because the manure is not thoroughly decomposed. The left graph is rape cultivated by using the incompletely decomposed material mixed soil, and the right graph is rape cultivated by using the completely decomposed material mixed soil.
After 2 weeks, 30 plants were randomly selected and measured for plant height, and as shown in fig. 2, the average plant height of the rapes grown by using the incompletely decomposed material mixed soil was 2.72 cm; the average plant height of the rape which is thoroughly decomposed by the product and mixed with soil for seedling culture is 5.01 cm.
Adding the incompletely decomposed feces and urine into the product, mixing, standing for 5 days, and repeating the above experiment, wherein the left diagram is an experimental group and the right diagram is a control group.
After 2 weeks, 30 plants were randomly selected and the plant height was measured, as shown in FIG. 3. The average plant height of the rape seedlings cultured by the thoroughly decomposed material mixed with the soil is 7.24 cm; the average height of the rape seedlings cultured by the control group by using the incompletely decomposed materials mixed with the soil is 4.93 cm.
The above description is only exemplary of the preferred embodiment of the present invention, and any modification, equivalent replacement, and improvement made within the spirit and scope of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A microbial deodorant liquid for public environments is characterized by comprising the following components in percentage by mass:
the balance of photosynthetic bacteria liquid, and the sum of the total mass percentage is 100%.
2. The microbial deodorant liquid for public environments as claimed in claim 1, which comprises the following components in percentage by mass:
the balance of photosynthetic bacteria liquid, and the sum of the total mass percentage is 100%.
3. A preparation method of microbial deodorant liquid for public environment is characterized by sequentially comprising the following steps:
(1) respectively preparing bacillus subtilis powder, bacillus licheniformis powder, saccharomyces cerevisiae powder, lactobacillus powder, trichoderma viride powder, aspergillus niger powder and photosynthetic bacteria liquid;
(2) respectively weighing bacillus subtilis powder, bacillus licheniformis powder, saccharomyces cerevisiae powder, lactobacillus powder, trichoderma viride powder, aspergillus niger powder and photosynthetic bacteria liquid according to the mass percentage;
(3) adding the bacillus subtilis powder and the bacillus licheniformis powder weighed in the step (2) into the photosynthetic bacteria liquid, and stirring while adding to obtain a mixture A;
(4) sequentially adding the saccharomyces cerevisiae powder and the lactobacillus powder weighed in the step (2) into the mixture A prepared in the step (3), stirring and adding the mixture A, and fully and uniformly mixing to obtain a mixture B;
(5) slowly adding the trichoderma viride powder and the aspergillus niger powder weighed in the step (2) into the mixture B prepared in the step (4), stirring and adding the mixture B, and fully and uniformly stirring to obtain a mixture C;
(6) filtering with a 60-mesh nylon mesh, removing impurities, detecting the technical indexes of the product, and subpackaging to obtain the microbial deodorant liquid.
4. The method for preparing microbial deodorant solution for public environment as claimed in claim 3, wherein the effective viable count of the final product of Bacillus subtilis powder of step (1) is not less than 5 x 1010The water content is 8-9% per gram.
5. The method for preparing microbial deodorant liquid for public environment as claimed in claim 3, wherein the effective viable count of the final product of the powder of Bacillus licheniformis in step (1) is not less than 5 x 1010The water content is 8-9% per gram.
6. The method for preparing microbial deodorant solution for public environment as claimed in claim 3, wherein the effective viable count of the Saccharomyces cerevisiae powder final product in the step (1) is not less than 2 x 1010The water content is 8-9% per gram.
7. The method as claimed in claim 6, wherein the powder of Lactobacillus is a commercially available agricultural grade fermented product of Lactobacillus plantarum, and the number of viable bacteria in the product is not less than 5X 1010The water content is 8-9% per gram.
8. The method for preparing a microbial deodorant solution for public environments as claimed in claim 3, wherein the method for preparing the Trichoderma viride powder in the step (1) comprises:
the method for separating the trichoderma viride adopts a dilution coating flat plate method, collects padding materials and rotten forage grass of a farm colony respectively, uniformly mixes the padding materials and the rotten forage grass according to the mass ratio of 1:1, weighs 10g of the rotten forage grass, adds the 10g of the rotten forage grass into 90mL of sterile water,placing on a shaking table with the rotation number of 180r/min, shaking for 1h, standing, sucking 100 mu L of supernatant, uniformly coating on a trichoderma screening culture medium plate, wherein the trichoderma screening culture medium is prepared according to the following method: adding 100g of fresh potato blocks into 1l of pure water, decocting for 10-15 min, cooling, homogenizing, adding pure water to 1000ml, sequentially adding 20g of brown sugar and KH2PO4 1.2g，MgSO40.6g, 0.008g of thiamine, 0.1g of chloramphenicol and 20g of agar, wherein the pH value before sterilization is 6.5-6.8, the mixture is placed in an incubator at 28 ℃ for culturing for 72 hours, and the colony morphology is observed;
the trichoderma viride is purified by adopting a single spore colony method, namely when trichoderma colonies grow on a plate and generate green spores, gently picking the spores by using an inoculation shovel, uniformly inoculating the spores on a trichoderma enhanced culture medium plate, wherein the trichoderma enhanced culture medium is prepared by the following method: adding 100g of fresh potato blocks into every 1L of soil leaching solution, decocting for 10-15 min, cooling, homogenizing, supplementing pure water to 1000mL, sequentially adding 20g of brown sugar and KH2PO4 1.2g，MgSO40.6g, 0.008g of thiamine, 0.1g of chloramphenicol and 20g of agar, wherein the pH value before sterilization is 6.5-6.8, the mixture is placed in an incubator at 28 ℃ for culturing for 72 hours, and the mixture is purified for multiple times by the method so as to obtain a plurality of single colonies with single morphology;
selecting the trichoderma spores obtained through purification, and carrying out microscopic morphological identification;
after identification, transferring the purified trichoderma viride spores into a test tube inclined plane of a trichoderma enhanced culture medium, culturing for 72 hours at 28 ℃ until hyphae grow on the whole inclined plane and mature spores are generated, and continuously performing propagation by using the method, namely primary strain culture;
selecting mature first-class seeds, scraping spores on the inclined plane of the test tube by using an inoculating shovel under aseptic condition, and inoculating the spores into a 500mL conical flask filled with a mold amplification culture medium, wherein the mold amplification culture medium comprises the following components: 32.5g of bran, 25g of corn flour, 7.5g of rice hull, 5g of soybean peptone or fish peptone, 42-47% of water content, 6.5-6.8 of pH before sterilization, culturing for 72h at 28 ℃ until culture medium materials are full of trichoderma viride conidia, sampling and counting, wherein the number of the trichoderma viride conidia is more than or equal to 5 multiplied by 109Per gram;
three-stage solid fermentation method of Trichoderma virideThe deep ventilation fermentation method is characterized in that the sterilized and cooled solid fermentation material and the secondary strain are uniformly mixed according to the mass ratio of 1:8, and the solid fermentation material comprises the following components: 3.25kg of bran, 2.5kg of corn flour, 0.75kg of rice hull, 0.5kg of soybean peptone or fish peptone and KH2PO4 1g，MgSO40.5g, the water content is 42-47%, the pH value before sterilization is 6.5-6.8, the mixture is flatly paved in a ventilation fermentation tank, the material depth is 15-20 cm, the fermentation is carried out for 72 hours at 28 ℃, and when the material core temperature is more than or equal to 40 ℃, the material turning operation is carried out once;
after fermentation, drying, crushing, sieving with a 60-mesh sieve, wherein the number of conidia of the Trichoderma viride in the product is more than or equal to 5 multiplied by 109The water content is 8-9% per gram.
9. The method for preparing the microbial deodorant liquid for public environments as claimed in claim 3, wherein the method for preparing the Aspergillus niger bacterial powder in the step (1) is specifically as follows:
aspergillus niger separation adopts a dilution coating flat plate method, surface soil of an organic fertilizer decomposition field is respectively collected, 10g of the organic fertilizer decomposition field is weighed and added into 90mL of sterile water, the sterile water is placed on a shaking table with the rotation number of 180r/min for shaking for 1h, standing is carried out, 100 mu L of supernate is absorbed and is uniformly coated on an Aspergillus niger selective culture medium flat plate, and the Aspergillus niger selective culture medium comprises the following components: (NH)4)2SO40.5g/L, 150g/L brown sugar and KH2PO4 1g/L，MgSO40.5g/L, 0.1g/L chloramphenicol, 30g/L agar, pH 6.5-6.8 before sterilization, culturing in an incubator at 28 deg.C for 72h, and observing colony morphology;
the aspergillus niger is purified by adopting a monospore colony method, namely when an aspergillus niger colony grows on a flat plate and generates black spores, the spores are lightly picked by using an inoculation shovel, evenly inoculated on the aspergillus niger selective culture medium flat plate and cultured in an incubator at 28 ℃ for 72 hours, and the purification is carried out for a plurality of times according to the method so as to obtain a plurality of single colonies with single shape;
selecting the purified aspergillus niger spores, and carrying out microscopic morphological identification;
after identification, transferring the purified aspergillus niger spores into an aspergillus niger selective culture medium test tube inclined plane, culturing for 72 hours at 28 ℃ until the whole inclined plane is full of hyphae and mature conidia are generated, and continuously performing propagation by using the method, namely primary strain culture;
selecting mature first-class seeds, scraping spores on the inclined plane of the test tube by using an inoculating shovel under aseptic condition, and inoculating the spores into a 500mL conical flask filled with a mold amplification culture medium, wherein the mold amplification culture medium comprises the following components: 32.5g of bran, 25g of corn flour, 7.5g of rice hull, 5g of soybean peptone or fish peptone, 42-47% of water content, 6.5-6.8 of pH before sterilization, culturing for 72h at 28 ℃ until the culture medium is full of Aspergillus niger conidia, sampling and counting, wherein the number of Aspergillus niger conidia is more than or equal to 5 multiplied by 109Per gram;
the three-stage solid fermentation of the aspergillus niger adopts a deep ventilation fermentation method, the sterilized and cooled solid fermentation material and the secondary strain are uniformly mixed according to the mass ratio of 1:8, and the solid fermentation material comprises the following components: 3.25kg of bran, 2.5kg of corn flour, 0.75kg of rice hull, 0.5kg of soybean peptone or fish peptone and KH2PO4 1g，MgSO40.5g, the water content is 42-47%, the pH value before sterilization is 6.5-6.8, the mixture is flatly paved in a ventilation fermentation tank, the material depth is 15-20 cm, the fermentation is carried out for 72 hours at 28 ℃, and when the material core temperature is more than or equal to 40 ℃, the material turning operation is carried out once;
after fermentation, drying, crushing and sieving with a 60-mesh sieve, wherein the number of aspergillus niger conidia in the product is more than or equal to 5 multiplied by 109The water content is 8-9% per gram.
10. The method for preparing microbial deodorant liquid for public environment as claimed in claim 3, wherein the method for preparing photosynthetic bacteria liquid in step (1) is specifically:
(1) separately weighing NaHCO31g, sodium acetate 1g, NaCl1g, NH4Cl 1g、MgSO4 0.3g、K2HPO40.2g, 0.2g of sodium propionate, 0.25g of malic acid, 0.2g of peptone, 0.1g of yeast extract and 1000mL of deionized water;
(2) adding the yeast extract and peptone weighed in the step (1) into deionized water, heating, and uniformly stirring to obtain a mixture D;
(3) weighing NaHCO in the step (1)3Sodium acetate, NaCl, NH4Cl、MgSO4、K2HPO4Sequentially adding sodium propionate and malic acid into the mixture D prepared in the step (2), and fully and uniformly mixing to obtain a mixture E;
(4) respectively weighing 10g of gamma-aminobutyric acid and 0.2g of glutamic acid, and dissolving in 1L of deionized water to obtain a mixture F;
(5) adding 1mL of mixture F into each liter of mixture E, adding the mixture F prepared in the step (4) into the mixture E prepared in the step (3), and fully and uniformly stirring to obtain a mixture G;
(6) collecting water samples on the surface layer of the breeding wastewater or the septic tank, and according to the water samples: and (3) fully mixing the mixture G in a volume ratio of 1: 6-1: 8, subpackaging the mixture in a transparent container, and performing light anaerobic culture under the following conditions: stirring or shaking regularly at 28-32 ℃ and 4000Lux, and culturing for 7-10 days.
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Citations (5)
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| CN1861199A (en) * | 2005-05-12 | 2006-11-15 | 中国农业大学 | Biological deodorization and purification agent, and its use |
| CN101186879A (en) * | 2007-12-05 | 2008-05-28 | 中国科学院南京土壤研究所 | Agriculture castoff compost ternary microorganism composite microbial inoculum |
| WO2008136545A1 (en) * | 2007-05-04 | 2008-11-13 | Pyung Kang Special Vehicle Co., Ltd. | Bacillus licheniformis bc4 kccm 10860p reducing nasty odor and heavy metal and clarification methods for resource recovery from food waste or livestock waste water by using it |
| CN104307014A (en) * | 2014-09-26 | 2015-01-28 | 任杰 | Microbial deodorant and preparation method thereof |
| CN104667320A (en) * | 2015-03-04 | 2015-06-03 | 蒋常德 | Composite microbial deodorant for treating household garbage and preparation method of deodorant |
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- 2021-10-15 CN CN202111204358.2A patent/CN113980840A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1861199A (en) * | 2005-05-12 | 2006-11-15 | 中国农业大学 | Biological deodorization and purification agent, and its use |
| WO2008136545A1 (en) * | 2007-05-04 | 2008-11-13 | Pyung Kang Special Vehicle Co., Ltd. | Bacillus licheniformis bc4 kccm 10860p reducing nasty odor and heavy metal and clarification methods for resource recovery from food waste or livestock waste water by using it |
| CN101186879A (en) * | 2007-12-05 | 2008-05-28 | 中国科学院南京土壤研究所 | Agriculture castoff compost ternary microorganism composite microbial inoculum |
| CN104307014A (en) * | 2014-09-26 | 2015-01-28 | 任杰 | Microbial deodorant and preparation method thereof |
| CN104667320A (en) * | 2015-03-04 | 2015-06-03 | 蒋常德 | Composite microbial deodorant for treating household garbage and preparation method of deodorant |
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