CN104478600A - Functional microbial agent and preparation method thereof - Google Patents
Functional microbial agent and preparation method thereof Download PDFInfo
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- CN104478600A CN104478600A CN201410786343.5A CN201410786343A CN104478600A CN 104478600 A CN104478600 A CN 104478600A CN 201410786343 A CN201410786343 A CN 201410786343A CN 104478600 A CN104478600 A CN 104478600A
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/90—Apparatus therefor
- C05F17/964—Constructional parts, e.g. floors, covers or doors
- C05F17/971—Constructional parts, e.g. floors, covers or doors for feeding or discharging materials to be treated; for feeding or discharging other material
- C05F17/979—Constructional parts, e.g. floors, covers or doors for feeding or discharging materials to be treated; for feeding or discharging other material the other material being gaseous
Abstract
The invention relates to a functional microbial agent and a preparation method thereof. A seed tank (6), a fermentation tank (5), a storage tank (9) and a metering filling tank (10) serving as main devices, a steam engine (2) and a steam filter (16) serving as heating sterilization devices, an air compressor (8), a gas storage tank (7), a freezing dryer (12), an oil-water separator (11), an air coarse filter (13), a pressure-reducing valve (14) and an air fine filter (15) serving as sterile air supply devices and a purified water tank (1), a circulating water tank (4) and a water pump (B) serving as circulating constant-temperature devices are connected through sealing pipelines to constitute a production line, technical processes, namely, an empty fermentation tank disinfection and liquid-added fermentation tank disinfection are carried out and a strain(A) is added into a culture solution in the seed tank (6) and is subjected to technical processes, namely, an inoculation and expanding propagation step and a storage filling step to prepare the functional microbial agent finished product. The functional microbial agent has the characteristics that the fertilizer efficiency is high, the microbial agent can promote plants to prevent insect pests diseases, the soil structure is improved and the microbial agent is non-toxic and harmless and is conductive to environment friendliness, the sustainable and long-term development of the farmland is promoted and increases in both production and income are promoted.
Description
Technical field
The present invention relates to plant microbial inoculum, particularly relate to the preparation method of the microbiobacterial agent promoting crop growth.
Background technology
China is the large agricultural country of 1,300,000,000 populations, with the arable land of Jin Zhan world cultivated area 7%, has supported the population of 20% in the world.Although make outstanding achievements, but pay heavy cost.The agricultural year of China is based upon a large amount of use on the basis of agriculture chemical to obtain, and the cultivated area of 7% consumes the chemical fertilizer of in the world 30%, makes the agroecological environment of China face unprecedented injury.A large amount of use chemical fertilizer causes soil nutrient structural imbalance, soil compaction, soil fertility to decline; The quality of farm crop declines, production cost is high, decline in benefits; Groundwater pollution causes body eutrophication, water environment degradation, the series of problems such as topsoil.A large amount of uses of agricultural chemicals, cause vast rural area Sichuan, add that Pesticide use ratio is unreasonable and some cash crop Pesticide use frequent, excess, severe contamination is caused to soil, surface water, underground water, a stepping of going forward side by side enters food chain, threatens to environmental organism and human health.Life-time service agricultural chemicals can cause the adaptability of environment to develop.Many insects develop immunity to drugs gene, and then cause the usage quantity of agricultural chemicals to increase further, increase further the harm of environment and the mankind, thus form vicious cycle.Along with the raising that international community requires agricultural product quality, China is caused to face export of farm produce difficulty, improve the ecotope of agricultural, the usage quantity of agriculture chemical must be reduced as far as possible, therefore microbial function microbial inoculum is used in research becomes key subjects, and fungus treatment becomes the only way of agriculture production.Current microbial inoculum application development is very fast, and very different, although some bacteria containing amount is many, miscellaneous bacteria rate is high, and quality is stable not, and effect is not ideal, falls flat in actual use.In order to meet growing demand, improving the performance quality of microbiobacterial agent further, popularizing the ecological agriculture in an all-round way, becoming following main development direction.
Summary of the invention
Object of the present invention, be just to provide a kind of fertilizer efficiency high, function is many, applied widely, profitable, nontoxic microbiobacterial agent, has the function of prevention and elimination of disease and pests, improvement soil, plays the effect being beneficial to environmental protection Sustainable development, promote that increasing crop yield increases income.
Task of the present invention completes like this: a kind of microbial function microbial inoculum and preparation method thereof is cultivated in research, with seeding tank, fermentor tank, hold-up vessel, metering filling tank is major equipment, with steamer, vapor filter is thermal sterilization equipment, with air compressor machine, gas-holder, lyophilizer, water-and-oil separator, air strainer, reducing valve, air strainer is sterile air air feed equipment, with pure water box, cyclic water tank, water pump is circulation constant temperature equipment, production line is connected and composed by sealing-duct, to disappear operation through sky, after the technological process of the real operation that disappears of liquid feeding, bacterial classification is added in the nutrient solution in seeding tank, numerous operation is expanded again through inoculation, store the technological process of filling operation, make microbial function microbial inoculum finished product.The disappear technological process of operation of sky is: closed by the air door of seeding tank and fermentor tank, open the down blow valve of air strainer, drain cold water, open steam valve, the pressure of air strainer is adjusted to 0.11 ~ 0.12MPa, 30 ~ 40min is kept to carry out steam sterilizing, steam off valve again, open air door, air flow is adjusted to 30L/min, pressure is 0.05MPa, ventilate 15 ~ 20min, then air door is closed, the pipeline internal gas pressure keeping sterile air pipe to be communicated with is malleation, by seeding tank (6) and fermentor tank venting port valve open, and open chuck wash water valve, logical steam 5min-10min in chuck, after cold water in emptying chuck, close chuck steam valve, then logical steam in seeding tank (6) and fermentor tank, in tank, temperature rises to 120 ~ 121 DEG C, turn vapor cock down, make pressure increase to 0.1 ~ 0.12MPa, turn steam valve down again, pressure-stabilisation is at 0.11 ~ 0.12MPa, keep 30 ~ 40min, temperature declines, continue to close each road steam valve, open large vent valve, Pressure Drop is to 0.03MPa, open air door and pass into sterile air, tank internal pressure rises to 0.05 ~ 0.1MPa, keep stable, when temperature is down to 40 DEG C, close air door, open drain tap venting, tank internal pressure reduces to 0, inserts pH electrode and Do electrode in tank.The technological process of the real operation that disappears of liquid feeding is: the nutrient solution prepared in culture tank is added space to respective 60% in seeding tank and fermentor tank respectively by nutrient solution pipe, in seeding tank, add defoamer 30 ml again, in fermentor tank, add defoamer 200 ~ 300ml, then in chuck, logical steam carries out preheating, start agitator, stir 25 ~ 40min, temperature rises to more than 80 DEG C, stop stirring and preheating, in tank, logical steam is to pressure 0.1 ~ 0.11MPa, temperature 120 ~ 121 DEG C, keep 30 ~ 40min, stop logical steam, Pressure Drop is to 0.03MPa; Open air door, pass into sterile air, pressure rises to 0.1 ~ 0.11MPa, opens vent valve and keeps pressure-stabilisation; Close chuck wash water valve, logical constant temp cooling water in chuck, temperature reduces to 40 DEG C, enters inoculation and expands numerous operation.The nutrient solution prepared in culture tank is: according to 1 part, weighing scale bean sprouts: the ratio that water is 5 parts adds water, adds bean sprouts in culture tank, electrified regulation is to boiling, powered-down after 40 minutes is boiled in boiling, leach bean sprouts, leave bean sprouts water in culture tank, in the water of bean sprouts, add the brown sugar of bean sprouts weight 10%, mix, Temperature fall to 60 ~ 70 DEG C, make nutrient solution.The technological process that numerous operation is expanded in inoculation is: the intake valve regulating seeding tank, reduce air input, by the Pressure Drop of seeding tank to below 0.01MPa, take off opening for feed lid, being soaked in concentration is in the alcohol of 75%, aseptic technique imports bacterial classification rapidly in seeding tank, screwing a lid on, pass into sterile air, is 30L/min in air flow quantity seeding tank, pressure rises to 0.05MPa, and arranging temperature in tank is 39.5 ~ 40.5 DEG C, agitator speed 180 ~ 200r/min; After 16h, every 2h sampling observation once, and after checking thalline to enter late log phase to 16 ~ 24h, the bacterium liquid in seeding tank is imported in fermentor tank by transfer pipet and proceeds to expand numerous cultivation; Bacterium liquid in seeding tank shuts air door, pressure release after deriving; In seeding tank, add water to 80%, blowing air, open agitator 20min, discharge water, clear up discharge nozzle simultaneously, repeat aforementioned sky and to disappear step.Bacterial classification be Bacillus licheniformis (
bacillus licheniformis), its preparation process takes from the stalk and corn cob that so become thoroughly decomposed, the mixture 100g synthesized according to the ratio of stalk 4 parts, corn cob 1 part, insert in 250ml Erlenmeyer flask, bottleneck adds tampon shutoff, is placed in 40 ° of C incubators, cultivate one week, form mixture culture sample; Then adopt serial dilutions to be coated with the mixture culture sample of enrichment, serial dilutions process is: get mixture culture sample 10g and join in the Erlenmeyer flask of splendid attire 90ml sterilized water, vibration mixes, and is fused in sterilized water, and dilution is 10
-1, obtain a diluent; From a diluent, get 1ml again, be moved in the test tube of splendid attire 9ml sterilized water, vibration mixes, and dilution is 10
-2, form secondary dilution liquid; From secondary dilution liquid, get 1ml again, be moved in the test tube of splendid attire 9ml sterilized water, vibration mixes, and dilution is 10
-3, form three diluents ... so successively analogize, repeat above dilution, until be diluted to 10
-6, form six diluents; Obtained containing stoste 10
-6the bacteria suspension of bacterium amount; Then stoste, content 10 is got
-4four diluents, 10
-5five diluents, 10
-6bacteria suspension, respectively get 0.1ml, be coated onto respectively cover substratum flat board on, in high-temperature cultivation case, cultivate 48h; Then take out flat board, single bacterium colony that picking comes in every shape 6, carries out line purifying respectively, then inserts in high-temperature cultivation case and cultivate 48h; And then inspection by sampling, select wherein to grow the fastest bacterial strain as bacterial classification, load Erlenmeyer flask.Substratum is inserted in heating container by mixture culture sample 200g, adds water boil, mix, and gets filtrate 300 ml, add 2% agar, be prepared from after autoclave sterilization according to mass ratio.The numerous culturing process of expansion of fermentor tank is: the bacterium liquid in seeding tank is imported in fermentor tank by transfer pipet, and fermentor tank arranges agitator speed 180 ~ 200r/min, and in tank, temperature is 39.5 ~ 40.5 DEG C, and fermentor tank air flow amount is 15-18m
3/ h, pressure remains on 0.05Mpa, after 20h, rotating speed is turned down to 150r/min, Do value maintenance more than 50,24h, every 2 ~ 4h sampling inspection once, when checking gemma rate to reach 80 ~ 90% to 24 ~ 48h, stop stirring, stopping air feed, under packing state, the bacterium liquid formed in fermentor tank passes through into liquid pipe and squeezes in hold-up vessel, enters and stores filling operation.The technological process storing filling operation is: by becoming liquid pipe to squeeze in the hold-up vessel of sterilization by the bacterium liquid in fermentor tank, normal temperature storage; Again by bacterium liquid by bacterium liquid pipe squeeze into metering filling tank (in, stir, namely make microbial function microbial inoculum finished product; Finally carry out metering filling input pod, with sealing machine sealing, after the assay was approved, joint sealing, warehouse-in.Microbial function microbial inoculum finished product is liquid state, and inspecting standard index is: liquid color is tawny, free from extraneous odour: pH value is pH value is 6.6 ~ 7.2, and gemma rate reaches 80 ~ 90%, and bacteria containing amount is 14 ~ 1,700,000,000/ml.
The gemma formed in the microbial inoculum that the present invention makes, acid-fast alkali-proof, high-low temperature resistant, survival ability is strong, the flora being in not eye state after being manured into soil can rapid series of expansion, increase the quantity of soil profitable strain, can the effective regulating plant rhizosphere soil microorganism eubiosis, promote other biological generation antibacterial substance, kill phytopathogen, the growth-promoting substance such as growth hormone material and Plant hormones regulators,gibberellins material can be secreted, extraneous root strong sprout, strengthen plant drought-resistant ability, can promote that soil aggregate is formed, and keeps soil from packing together, prevent the moisture in soil from evaporating.The output of farm crop can be increased, effectively prevention and elimination of disease and pests.
Carry out implementing, testing according to technique scheme, prove that the present invention has fertilizer efficiency high, plant preventing disease and pest can be promoted, improvement Soil structure, nontoxic, be beneficial to the feature of environmental protection, can promote that farmland continues long-acting development, promote that increasing crop yield increases income, function is many, cost is low, there are good economic results in society, reach predetermined object preferably.
Accompanying drawing explanation
Accompanying drawing is apparatus for production line connection diagram of the present invention.
In figure, 1-pure water pot, 2-steamer.3-culture tank, 4-cyclic water tank, 5-fermentor tank, 6-seeding tank, 7-gas-holder, 8-air compressor machine, 9-hold-up vessel, 10-metering filling tank, 11-water-and-oil separator, 12-lyophilizer, 13-air strainer, 14-reducing valve, 15-air strainer, 16-vapor filter; A-bacterial classification, B-water pump, C-discharge nozzle, D-bean sprouts, G-pure water pipe, H-return water pipe, J-under meter, L-transfer pipet, M-nutrient solution pipe, P-vapor pipe, Q-sterile air pipe, R-vapour pipe, S-thief hole, T-brown sugar, W-one-tenth liquid pipe, X-constant temperature water pipe, Y-bacterium liquid pipe, Z-container.
Embodiment
Below in conjunction with accompanying drawing, the specific embodiment of the present invention is described.
Accompanying drawings, the present invention is with seeding tank 6, fermentor tank 5, hold-up vessel 9, metering filling tank 10 is major equipment, with steamer 2, vapor filter 16 is thermal sterilization equipment, with air compressor machine 8, gas-holder 7, lyophilizer 12, water-and-oil separator 11, air strainer 13, reducing valve 14, air strainer 15 is sterile air air feed equipment, with pure water box 1, cyclic water tank 4, water pump B is circulation constant temperature equipment, production line is connected and composed by sealing-duct, to disappear operation through sky, after the technological process of the real operation that disappears of liquid feeding, bacterial classification A is added in the nutrient solution in seeding tank 6, numerous operation is expanded again through inoculation, store the technological process of filling operation, make microbial function microbial inoculum finished product.
The disappear technological process of operation of sky is: closed by the air door of seeding tank 6 and fermentor tank 5, open the down blow valve of air strainer 15, drain cold water, open steam valve, the pressure of air strainer 15 is adjusted to 0.11 ~ 0.12MPa, 30 ~ 40min is kept to carry out steam sterilizing, steam off valve again, open air door, air flow is adjusted to 30L/min, pressure is 0.05MPa, ventilate 15 ~ 20min, then air door is closed, the pipeline internal gas pressure keeping sterile air pipe Q to be communicated with is malleation, by seeding tank 6 and fermentor tank 5 venting port valve open, and open chuck wash water valve, logical steam 5min-10min in chuck, after cold water in emptying chuck, close chuck steam valve, then logical steam in seeding tank 6 and fermentor tank 5, in tank, temperature rises to 120 ~ 121 DEG C, turn vapor cock down, make pressure increase to 0.1 ~ 0.12MPa, turn steam valve down again, pressure-stabilisation is at 0.11 ~ 0.12MPa, keep 30 ~ 40min, temperature declines, continue to close each road steam valve, open large vent valve, Pressure Drop is to 0.03MPa, open air door and pass into sterile air, tank internal pressure rises to 0.05 ~ 0.1MPa, keep stable, when temperature is down to 40 DEG C, close air door, open drain tap venting, tank internal pressure reduces to 0, inserts pH electrode and Do electrode in tank.
The technological process of the real operation that disappears of liquid feeding is: the nutrient solution prepared in culture tank 3 is added space to respective 60% in seeding tank 6 and fermentor tank 5 respectively by nutrient solution pipe M, defoamer 30 ml is added again in seeding tank 6, defoamer 200 ~ 300ml is added in fermentor tank 5, then in chuck, logical steam carries out preheating, start agitator, stir 25 ~ 40min, temperature rises to more than 80 DEG C, stop stirring and preheating, in tank, logical steam is to pressure 0.1 ~ 0.11MPa, temperature 120 ~ 121 DEG C, keep 30 ~ 40min, stop logical steam, Pressure Drop is to 0.03MPa, open air door, pass into sterile air, pressure rises to 0.1 ~ 0.11MPa, opens vent valve and keeps pressure-stabilisation, close chuck wash water valve, logical constant temp cooling water in chuck, temperature reduces to 40 DEG C, enters inoculation and expands numerous operation.
The nutrient solution prepared in described culture tank 3 is: according to weighing scale bean sprouts D1 part: the ratio that water is 5 parts adds water, adds bean sprouts D in culture tank 3, electrified regulation is to boiling, and powered-down after 40 minutes is boiled in boiling, leaches bean sprouts D, bean sprouts water is left in culture tank 3, boiling powered-down after 40 minutes to rising, leaching bean sprouts D, in culture tank 3, leaving bean sprouts water, the brown sugar T of bean sprouts D weight 10% is added in the water of bean sprouts, mix, Temperature fall to 60 ~ 70 DEG C, make nutrient solution.
The technological process that numerous operation is expanded in inoculation is: the intake valve regulating seeding tank 6, reduce air input, by the Pressure Drop of seeding tank 6 to below 0.01MPa, take off opening for feed lid, being soaked in concentration is in the alcohol of 75%, aseptic technique imports bacterial classification A rapidly in seeding tank 6, screwing a lid on, pass into sterile air, is 30L/min in air flow quantity seeding tank 6, pressure rises to 0.05MPa, and arranging temperature in tank is 39.5 ~ 40.5 DEG C, agitator speed 180 ~ 200r/min; After 16h, every 2h sampling observation once, and after checking thalline to enter late log phase to 16 ~ 24h, the bacterium liquid in seeding tank 6 is imported in fermentor tank 5 by transfer pipet L and proceeds to expand numerous cultivation; Bacterium liquid in seeding tank 6 shuts air door, pressure release after deriving; In seeding tank 6, add water to 80%, blowing air, open agitator 20min, discharge water, clear up discharge nozzle C simultaneously, repeat aforementioned sky and to disappear step.
Aforementioned inoculation is expanded in numerous operation, the bacterial classification A of described importing seeding tank 6 be Bacillus licheniformis (
bacillus licheniformis), its preparation process is: choose stalk and corn cob that nature becomes thoroughly decomposed, the mixture 100g synthesized according to the ratio of stalk 4 parts, corn cob 1 part, insert in 250ml Erlenmeyer flask, bottleneck adds tampon shutoff, is placed in 40 ° of C incubators, cultivate one week, form mixture culture sample; Then adopt serial dilutions to be coated with the mixture culture sample of enrichment, serial dilutions process is: get mixture culture sample 10g and join in the Erlenmeyer flask of splendid attire 90ml sterilized water, vibration mixes, and is fused in sterilized water, and dilution is 10
-1, obtain a diluent; From a diluent, get 1ml again, be moved in the test tube of splendid attire 9ml sterilized water, vibration mixes, and dilution is 10
-2, form secondary dilution liquid; From secondary dilution liquid, get 1ml again, be moved in the test tube of splendid attire 9ml sterilized water, vibration mixes, and dilution is 10
-3, form three diluents ... so successively analogize, repeat above dilution, until be diluted to 10
-6, form six diluents; Obtained containing stoste 10
-6the bacteria suspension of bacterium amount; Then choose: stoste, content 10
-4four diluents, 10
-5five diluents, 10
-6bacteria suspension, 0.1ml got by every sample, be coated onto respectively cover substratum flat board on, in high-temperature cultivation case, cultivate 48h; Then take out flat board, single bacterium colony that picking comes in every shape 6, observe under powerful microscope, colony characteristics: form is irregular, edge is irregular, and centre is slightly swelled, edge roughness, and centre is slightly smooth.Choose come in every shape 6 single bacterium colonies are carried out line purifying respectively, then inserts in high-temperature cultivation case and cultivate 48h; And then inspection by sampling, it is elongated rod shape that thalli morphology is observed in film-making, has gemma.Select wherein to grow the fastest bacterial strain as bacterial classification A, load in Erlenmeyer flask for subsequent use.
The substratum that flat board described in above-mentioned technological process covers is special processing preparation, is insert in heating container by mixture culture sample 200g, adds water boil, mix, get filtrate 300 ml, add 2% agar according to mass ratio, be prepared from after autoclave sterilization.
Described inoculation is expanded in numerous operation, and the bacterium liquid in seeding tank 6 is imported in fermentor tank 5 by transfer pipet L, proceeds to inoculate the numerous culture process process of expansion expanding numerous operation later stage.The numerous culturing process of expansion of fermentor tank 5 is: bacterium liquid imports in fermentor tank 5, and fermentor tank 5 arranges agitator speed 180 ~ 200r/min, and in tank, temperature is 39.5 ~ 40.5 DEG C, and fermentor tank 5 air flow amount is 15-18m
3/ h, pressure remains on 0.05Mpa, after 20h, rotating speed is turned down to 150r/min, after Do value maintenance more than 50,24h, every 2 ~ 4h sampling inspection once, when checking gemma rate to reach 80 ~ 90% to 24 ~ 48h, stop stirring, stopping air feed, under packing state, the bacterium liquid formed in fermentor tank 5 passes through into liquid pipe W and squeezes in hold-up vessel 9.Inoculation is expanded numerous operation and is completed, and enters and stores filling operation.
The technological process storing filling operation is: by becoming liquid pipe W to squeeze in the hold-up vessel 9 of sterilization by the bacterium liquid in fermentor tank 5, normal temperature storage; Again bacterium liquid is squeezed in metering filling tank 10 by bacterium liquid pipe Y, stir, namely make microbial function microbial inoculum finished product; Finally carry out metering filling input pod Z, with sealing machine sealing, after the assay was approved, joint sealing, warehouse-in.
The microbial function microbial inoculum finished product that the present invention makes is liquid state, and inspecting standard index is: color is tawny, free from extraneous odour; PH value is 6.6 ~ 7.2, and gemma rate reaches 80 ~ 90%, and bacteria containing amount is 14 ~ 1,700,000,000/ml.Thalli morphology is observed in film-making, and colony characteristics is: form is irregular, and edge is irregular, and centre is slightly swelled, edge roughness, and centre is slightly smooth; Thalli morphology is elongated rod shape, has gemma.
Adopt microbial inoculum of the present invention to farm crop carry out punching execute time, according to every mu of 2L microbial inoculum consumption, when irrigating, thin up 50 times, by dilution after bacterium liquid with water flood irrigation; When filling with root, by every mu of 1 ~ 2L microbial inoculum consumption, thin up 500 times, waters in crop root part by the bacterium liquid after dilution, determines the dilution bacterium liquid consumption of every strain plant according to different densities; When dipping in root, according to every mu of 1 ~ 2L microbial inoculum consumption, thin up 200 times, immerses root in dilution bacterium liquid, soaks 10 ~ 20min before seedling replanting; When seed dressing or seed soaking, 50 ml microbial inoculums and 10kg seed are puddled evenly.
The gemma formed in the microbial inoculum that the present invention makes, acid-fast alkali-proof, high-low temperature resistant, survival ability is strong, the flora being in not eye state after being manured into soil can rapid series of expansion, increase the quantity of soil profitable strain, can the effective regulating plant rhizosphere soil microorganism eubiosis, promote other biological generation antibacterial substance, kill phytopathogen, the growth-promoting substance such as growth hormone material and Plant hormones regulators,gibberellins material can be secreted, extraneous root strong sprout, strengthen plant drought-resistant ability, can promote that soil aggregate is formed, and keeps soil from packing together, prevent the moisture in soil from evaporating.For administering the soil-borne diseases such as root rot common in gourd, fruit and vegetable process of growth, damping-off, red root disease, blight, base rot disease, samping off, effect is fine.
The present invention have fertilizer efficiency high, plant preventing disease and pest, improvement Soil structure, nontoxic, the feature that is beneficial to environmental protection can be promoted, can promote that farmland continues long-acting development, promote that increasing crop yield increases income, function is many, cost is low, has good economic results in society and application value.
Claims (10)
1. microbial function microbial inoculum and preparation method thereof, it is characterized in that with seeding tank (6), fermentor tank (5), hold-up vessel (9), metering filling tank (10) is major equipment, with steamer (2), vapor filter (16) is thermal sterilization equipment, with air compressor machine (8), gas-holder (7), lyophilizer (12), water-and-oil separator (11), air strainer (13), reducing valve (14), air strainer (15) is sterile air air feed equipment, with pure water box (1), cyclic water tank (4), water pump (B) is circulation constant temperature equipment, production line is connected and composed by sealing-duct, to disappear operation through sky, after the technological process of the real operation that disappears of liquid feeding, bacterial classification (A) is added in the nutrient solution in seeding tank (6), numerous operation is expanded again through inoculation, store the technological process of filling operation, make microbial function microbial inoculum finished product.
2. according to microbial function microbial inoculum according to claim 1 and preparation method thereof, it is characterized in that the disappear technological process of operation of described sky is: closed by the air door of seeding tank (6) and fermentor tank (5), open the down blow valve of air strainer (15), drain cold water, open steam valve, the pressure of air strainer (15) is adjusted to 0.11 ~ 0.12MPa, 30 ~ 40min is kept to carry out steam sterilizing, steam off valve again, open air door, air flow is adjusted to 30L/min, pressure is 0.05MPa, ventilate 15 ~ 20min, then air door is closed, the pipeline internal gas pressure keeping sterile air pipe (Q) to be communicated with is malleation, by seeding tank (6) and fermentor tank (5) venting port valve open, and open chuck wash water valve, logical steam 5min-10min in chuck, after cold water in emptying chuck, close chuck steam valve, then logical steam in seeding tank (6) and fermentor tank (5), in tank, temperature rises to 120 ~ 121 DEG C, turn vapor cock down, make pressure increase to 0.1 ~ 0.12MPa, turn steam valve down again, pressure-stabilisation is at 0.11 ~ 0.12MPa, keep 30 ~ 40min, temperature declines, continue to close each road steam valve, open large vent valve, Pressure Drop is to 0.03MPa, open air door and pass into sterile air, tank internal pressure rises to 0.05 ~ 0.1MPa, keep stable, when temperature is down to 40 DEG C, close air door, open drain tap venting, tank internal pressure reduces to 0, inserts pH electrode and Do electrode in tank.
3. according to microbial function microbial inoculum according to claim 1 and preparation method thereof, it is characterized in that the technological process of the real operation that disappears of described liquid feeding is: the nutrient solution prepared in culture tank (3) to be added in seeding tank (6) and fermentor tank (5) respectively the space of extremely respective 60% by nutrient solution pipe (M), defoamer 30 ml is added again in seeding tank (6), defoamer 200 ~ 300ml is added in fermentor tank (5), then in chuck, logical steam carries out preheating, start agitator, stir 25 ~ 40min, temperature rises to more than 80 DEG C, stop stirring and preheating, in tank, logical steam is to pressure 0.1 ~ 0.11MPa, temperature 120 ~ 121 DEG C, keep 30 ~ 40min, stop logical steam, Pressure Drop is to 0.03MPa, open air door, pass into sterile air, pressure rises to 0.1 ~ 0.11MPa, opens vent valve and keeps pressure-stabilisation, close chuck wash water valve, logical constant temp cooling water in chuck, temperature reduces to 40 DEG C, enters inoculation and expands numerous operation.
4. according to microbial function microbial inoculum according to claim 3 and preparation method thereof, it is characterized in that the nutrient solution prepared in described culture tank (3) is: according to weighing scale bean sprouts (D) 1 part: the ratio that water is 5 parts adds water, adds bean sprouts (D) in culture tank (3), electrified regulation is to boiling, powered-down after 40 minutes is boiled in boiling, leach bean sprouts (D), culture tank leaves bean sprouts water in (3), the brown sugar (T) of bean sprouts (D) weight 10% is added in the water of bean sprouts, mix, Temperature fall to 60 ~ 70 DEG C, make nutrient solution.
5. according to microbial function microbial inoculum according to claim 1 and preparation method thereof, it is characterized in that the technological process that numerous operation is expanded in described inoculation is: the intake valve regulating seeding tank (6), reduce air input, by the Pressure Drop of seeding tank (6) to below 0.01MPa, take off opening for feed lid, being soaked in concentration is in the alcohol of 75%, aseptic technique imports rapidly bacterial classification (A) in seeding tank (6), screw a lid on, pass into sterile air, be 30L/min in air flow quantity seeding tank (6), pressure rises to 0.05MPa, arranging temperature in tank is 39.5 ~ 40.5 DEG C, agitator speed 180 ~ 200r/min, after 16h, every 2h sampling observation once, and after checking thalline to enter late log phase to 16 ~ 24h, the bacterium liquid in seeding tank (6) is imported in fermentor tank (5) by transfer pipet (L) and proceeds to expand numerous cultivation, bacterium liquid in seeding tank (6) shuts air door, pressure release after deriving, in seeding tank (6), add water to 80%, blowing air, open agitator 20min, discharge water, clear up discharge nozzle (C) simultaneously, repeat aforementioned sky and to disappear step.
6. according to microbial function microbial inoculum according to claim 5 and preparation method thereof, it is characterized in that described bacterial classification (A) for Bacillus licheniformis (
bacillus licheniformis), its preparation process is: take from the stalk and corn cob that so become thoroughly decomposed, the mixture 100g synthesized according to the ratio of stalk 4 parts, corn cob 1 part, insert in 250ml Erlenmeyer flask, bottleneck adds tampon shutoff, is placed in 40 ° of C incubators, cultivate one week, form mixture culture sample; Then adopt serial dilutions to be coated with the mixture culture sample of enrichment, serial dilutions process is: get mixture culture sample 10g and join in the Erlenmeyer flask of splendid attire 90ml sterilized water, vibration mixes, and is fused in sterilized water, and dilution is 10
-1, obtain a diluent; From a diluent, get 1ml again, be moved in the test tube of splendid attire 9ml sterilized water, vibration mixes, and dilution is 10
-2, form secondary dilution liquid; From secondary dilution liquid, get 1ml again, be moved in the test tube of splendid attire 9ml sterilized water, vibration mixes, and dilution is 10
-3, form three diluents ... so successively analogize, repeat above dilution, until be diluted to 10
-6, form six diluents; Obtained containing stoste 10
-6the bacteria suspension of bacterium amount; Then stoste, content 10 is got
-4four diluents, 10
-5five diluents, 10
-6bacteria suspension, respectively get 0.1ml, be coated onto respectively cover substratum flat board on, in high-temperature cultivation case, cultivate 48h; Then take out flat board, single bacterium colony that picking comes in every shape 6, carries out line purifying respectively, then inserts in high-temperature cultivation case and cultivate 48h; And then inspection by sampling, select wherein to grow the fastest bacterial strain as bacterial classification (A), load in Erlenmeyer flask for subsequent use.
7. according to microbial function microbial inoculum according to claim 6 and preparation method thereof, it is characterized in that described substratum is inserted in heating container by mixture culture sample 200g, add water boil, mix, get filtrate 300 ml, add 2% agar according to mass ratio, be prepared from after autoclave sterilization.
8. according to microbial function microbial inoculum according to claim 5 and preparation method thereof, it is characterized in that the numerous culturing process of expansion of described fermentor tank (5) is: the bacterium liquid in seeding tank (6) imports in fermentor tank (5) by transfer pipet (L), fermentor tank (5) arranges agitator speed 180 ~ 200r/min, in tank, temperature is 39.5 ~ 40.5 DEG C, and fermentor tank (5) air flow amount is 15-18m
3/ h, pressure remains on 0.05Mpa, after 20h, rotating speed is turned down to 150r/min, Do value maintenance more than 50,24h, every 2 ~ 4h sampling inspection once, when checking gemma rate to reach 80 ~ 90% to 24 ~ 48h, stop stirring, stopping air feed, under packing state, the bacterium liquid formed in fermentor tank (5) passes through into liquid pipe (W) and squeezes in hold-up vessel (9), enters and stores filling operation.
9. according to microbial function microbial inoculum according to claim 1 and preparation method thereof, it is characterized in that the technological process of the filling operation of described storage is: by becoming liquid pipe (W) to squeeze in the hold-up vessel (9) of sterilization by the bacterium liquid in fermentor tank (5), normal temperature storage; Again bacterium liquid is squeezed in metering filling tank (10) by bacterium liquid pipe (Y), stir, namely make microbial function microbial inoculum finished product; Finally carry out metering filling input pod (Z).
10., according to microbial function microbial inoculum according to claim 7 and preparation method thereof, it is characterized in that described microbial function microbial inoculum finished product is liquid state, inspecting standard index is: color is tawny, free from extraneous odour; PH value is 6.6 ~ 7.2, and gemma rate reaches 80 ~ 90%, and bacteria containing amount is 14 ~ 1,700,000,000/ml.
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| CN112494668A (en) * | 2020-12-17 | 2021-03-16 | 华帝股份有限公司 | Sterilization cooking control system and sterilization control method |
| CN117228819A (en) * | 2023-11-15 | 2023-12-15 | 江苏今石环境有限责任公司 | Heterotrophic nitrification and aerobic denitrification strain in-situ expansion culture system and method |
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