CN104862298A - Composite microbial culture starter and preparation method thereof - Google Patents
Composite microbial culture starter and preparation method thereof Download PDFInfo
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- CN104862298A CN104862298A CN201510196684.1A CN201510196684A CN104862298A CN 104862298 A CN104862298 A CN 104862298A CN 201510196684 A CN201510196684 A CN 201510196684A CN 104862298 A CN104862298 A CN 104862298A
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Abstract
The invention discloses a composite microbial culture starter and a preparation method thereof, and aims to solve the technical problem of efficient and comprehensive organic waste fermentation and composting processing. The composite microbial culture starter comprises a carrier and a compound strain, the carrier is natural zeolite powder, and the compound strain comprises bacillus subtilis, lactobacillus acidophilus, lactobacillus delbrueckii subspecies bulgaricus, saccharomyces cerevisiae, bacillus megatherium and clostridium pasteurianum. The preparation method includes inoculation, primary culture, secondary culture, mixing to obtain a compound bacterium fluid, adsorption of the compound microbial into the natural zeolite powder with the granularity of 200 mesh, and drying to obtain the composite microbial culture starter. Compared with the prior art, by efficient and comprehensive of the decompose organic wastes, the pollution caused by the organic wastes on the environment can be completely solved by the composite microbial culture starter, the resource properties of the organic wastes are improved, and the prepared multifunctional organic fertilizer provides comprehensive nutrients needed for plant growth, and improves the soil very well.
Description
Technical field
The present invention relates to a kind of organic fertilizer and preparation method thereof, particularly a kind of microbial starter culture and preparation method thereof of organic fertilizer.
Background technology
Day by day increase according to the organic solid castoff quantity that the agriculture industrialization in current cities and towns and the two-way Development patterns of urbanization produce, from the classification of agricultural structure aspect, the major source of agricultural wastes is from livestock industry, plant husbandry.On the other hand, town dweller's domestic refuse contains a large amount of organic waste, as changing food waste, sewage work every day the mud that produces.The organic waste that livestock industry, plant husbandry produce have recycles value, and wherein fowl and animal excrement contains the macroelement such as nitrogen, phosphorus, potassium needed for plant growth, again containing micro element in calcium, magnesium, sulphur, zinc, boron, molybdenum, copper, iron etc.The stalk, slag thing etc. of plant husbandry are containing a large amount of organic.The composition of changing food waste is based on degradable organism, and main component has starch contained by staple food, vegetables and the Mierocrystalline cellulose contained by plant stem-leaf, poly-pentose, the protein contained by meat and fat, monose, tartaric acid and pectin etc. contained by fruit.The mud that sewage work produces every day contains the macroelement such as nitrogen, phosphorus, potassium needed for plant growth and enriches organic.Nutrient contained by above organic waste all has recycles value, after inoculation fermentation, composting, can be made into fertilizer, namely can as the nutrient of plant-growth, Soil structure can be improved again, increase soil fertility, promote the growth of crop, but prior art is mixed with fertilizer after being fermented respectively by above-mentioned different classes of organic waste often again, not only function is few, fertilizer efficiency is low, and organic waste is made a low multiple use, can't effective kill harmful pathogenic bacteria, very smelly, unfavorable to people engaged in agriculture's health.
Summary of the invention
The object of this invention is to provide a kind of compound microbial culture starter and preparation method thereof, the technical problem that solve efficiently and comprehensively makes fermentation of organic wastes and composting, improves fertilizer efficiency, protecting agriculture practitioner health.
The present invention is by the following technical solutions: a kind of compound microbial culture starter, be made up of carrier and composite bacteria, mix with the mass ratio 1-3:1 of the composite bacteria liquid of composite bacteria by carrier, obtain after drying, described carrier is granularity 200 object natural zeolite powder, described composite bacteria liquid is made up of the mass percent of following bacterial classification bacterium liquid: subtilis 20-30%, Lactobacterium acidophilum 15-20%, lactobacillus delbruockii subspecies bulgaricus 15-20%, yeast saccharomyces cerevisiae 10-15%, bacillus megaterium 10-15%, Pasteur's Clostridium 10-15%, described each bacterial classification bacterium liquid every ml of carrier 2-3 hundred million bacterium number.
Composite bacteria liquid of the present invention is made up of the mass percent of following bacterium liquid: subtilis 25%, Lactobacterium acidophilum 15%, lactobacillus delbruockii subspecies bulgaricus 15%, yeast saccharomyces cerevisiae 15%, bacillus megaterium 15%, Pasteur's Clostridium 15%, described each bacterium liquid every ml of carrier 200,000,000 bacterium number.
A preparation method for compound microbial culture starter, comprises the following steps:
One, inoculate, respectively by subtilis, Lactobacterium acidophilum, lactobacillus delbruockii subspecies bulgaricus, yeast saccharomyces cerevisiae, bacillus megaterium and Pasteur's Clostridium, under room temperature, relative humidity is 50%, cultivate 16-20h, then respectively join in sterilized water, vibrate and obtain bacterium liquid after 30 minutes;
Two, one-level is cultivated, described each bacterium liquid is inoculated in the Erlenmeyer flask filling level liquid substratum respectively, inoculum size is 5%-10%, the pH value of substratum is 6.0-8.0,24-48h is cultivated with 180r/min rotating speed, obtain one-level to spread cultivation bacterium liquid, the one-level bacterium quantity in bacterium liquid that spreads cultivation reaches 2-3 hundred million/ml respectively;
Three, secondary is cultivated, each one-level being spread cultivation after bacterium liquid mixes with secondary liquid substratum is inoculated in the Erlenmeyer flask being loaded with secondary liquid substratum, inoculum size is 5%-10%, 48-72h is cultivated with 180r/min rotating speed, obtain secondary to spread cultivation bacterium liquid, the secondary bacterium quantity in bacterium liquid that spreads cultivation reaches 2-3 hundred million/ml respectively;
Four, by the spread cultivation mass percentage of bacterium liquid of each bacterial classification secondary be: subtilis 20-30%, Lactobacterium acidophilum 15-20%, lactobacillus delbruockii subspecies bulgaricus 15-20%, yeast saccharomyces cerevisiae 10-15%, bacillus megaterium 10-15%, Pasteur's Clostridium 10-15%, is mixed to get composite bacteria liquid; Composite bacteria liquid is adsorbed onto on granularity 200 object natural zeolite powder, granularity 200 object natural zeolite powder with have the mass ratio of the composite bacteria liquid of composite bacteria to be 1-3:1, dry, obtain compound microbial culture starter.
The step 2 inoculum size of the inventive method is 10%, and the pH value of described substratum is 6.0-7.0; Described with 180r/min rotating speed, cultivate 32h.
Bacterium quantity 200,000,000/ml respectively in the step 2 bacterium liquid of the inventive method.
The level liquid substratum composition of the step 2 of the inventive method is respectively:
(1) subtilis level liquid substratum consists of: peptone 5.0g, beef leaching thing 3.0g, NaCl 5.0g, agar 15.0g, distilled water 1.0L; Mix rear 112 DEG C of sterilizing 20min, naturally cool to room temperature, pH7.0;
(2) Lactobacterium acidophilum level liquid substratum consists of: casein peptone 10.0g, extractum carnis 10.0g, yeast powder 5.0g, glucose 5.0g, sodium acetate 5.0g, dibasic ammonium citrate 2.0g, sorbitan monooleate Soxylat A 25-7 1.0g, K
2hPO
42.0g, MgSO
4.7H
2o 0.2g, MnSO
4.H2O 0.05g, CaCO
320.0g, agar 15.0g, distilled water 1.0L; Mix rear 112 DEG C of sterilizing 20min, naturally cool to 37 DEG C, pH6.8;
(3) lactobacillus delbruockii subspecies bulgaricus level liquid substratum consists of: casein peptone 10.0g, extractum carnis 10.0g, yeast powder 5.0g, glucose 5.0g, sodium acetate 5.0g, dibasic ammonium citrate 2.0g, Tween 801.0g, K
2hPO
42.0g, MgSO
4.7H
2o 0.2g, MnSO
4.H
2o 0.05g, CaCO
320.0g, agar 15.0g, distilled water 1.0L; Mix rear 112 DEG C of sterilizing 20min, naturally cool to 37 DEG C, pH6.8;
(4) yeast saccharomyces cerevisiae level liquid substratum consists of: 5 ° of B é wort 1.0L, agar 15.0g; Mix rear 112 DEG C of sterilizing 20min, natural in 28 DEG C, natural pH;
(5) bacillus megaterium level liquid substratum consists of: peptone 5.0g, beef leaching thing 3.0g, NaCl 5.0g, agar 15.0g, distilled water 1.0L; Mix rear 112 DEG C of sterilizing 20min, naturally cool to 30 DEG C, pH7.0;
(6) Pasteur's Clostridium level liquid substratum consists of: peptone 10.0g, beef extract 3.0g, NaCl 5.0g, agar 15.0g, distilled water 1.0L; Mix rear 112 DEG C of sterilizing 20min, naturally cool to culture temperature 37 DEG C, pH7.0.
The step 3 inoculum size of the inventive method is 10%; Described with 180r/min rotating speed cultivation 48h.
The secondary of the step 3 of the inventive method bacterium quantity in bacterium liquid that spreads cultivation is respectively 200,000,000/ml.
The secondary liquid substratum of the step 3 of the inventive method consists of: peptone 10.0g, extractum carnis 10.0g, agar 15.0g, molasses 5.0g, MgSO
4.7H
2o 0.2g, MnSO
4.H
2o 0.05g, CaCO
320.0g, NaCl 5.0g, distilled water 1.0L; Mix rear 112 DEG C of sterilizing 20min, naturally cool to room temperature, natural pH.
The spread cultivation mass percentage of bacterium liquid of each bacterial classification secondary of step 4 of the inventive method is: subtilis 25%, Lactobacterium acidophilum 15%, lactobacillus delbruockii subspecies bulgaricus 15%, yeast saccharomyces cerevisiae 15%, bacillus megaterium 15%, Pasteur's Clostridium 15%.
The present invention compared with prior art, compound microbial culture starter can efficiently and comprehensively decompose organic waste, the pollution that thorough solution organic waste brings environment, improve the Resource Properties of organic waste, turn waste into wealth, through the Multifunctional organic fertilizer that compound microbial culture starter is made, the comprehensive nutrient needed for plant-growth is provided, and improves soil well.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail.Compound microbial culture starter of the present invention, is made up of carrier and composite bacteria, mixes, obtain after drying by carrier with the mass ratio 1-3:1 of the composite bacteria liquid of composite bacteria, each bacterial classification bacterium liquid every ml of carrier 2-3 hundred million bacterium number, preferably 200,000,000 bacterium numbers.Carrier is granularity 200 object natural zeolite powder.Bacterial classification is: subtilis (Bacillus substilis, strain number CICC23013, purchased from Chinese industrial Microbiological Culture Collection administrative center), Lactobacterium acidophilum (Lactobacillus acidophilus, strain number CICC22150, purchased from Chinese industrial Microbiological Culture Collection administrative center), lactobacillus delbruockii subspecies bulgaricus (Lactobacillus delbrueckiisubsp.bulgaricus, strain number CICC22153, purchased from Chinese industrial Microbiological Culture Collection administrative center), yeast saccharomyces cerevisiae (Saccharomyces cerevisiae, strain number CICC32390, purchased from Chinese industrial Microbiological Culture Collection administrative center), bacillus megaterium (Bacillus megaterium, strain number CICC21446, purchased from Chinese industrial Microbiological Culture Collection administrative center) and Pasteur's Clostridium (Clostridium pasteurianum, strain number CGMCC1.2675, purchased from China General Microbiological culture presevation administrative center).The mass percentage of each bacterial classification bacterium liquid of described composite bacteria liquid is: subtilis 20-30%, Lactobacterium acidophilum 15-20%, lactobacillus delbruockii subspecies bulgaricus 15-20%, yeast saccharomyces cerevisiae 10-15%, bacillus megaterium 10-15%, Pasteur's Clostridium 10-15%.The mass percentage of good each bacterial classification bacterium liquid is: subtilis 25%, Lactobacterium acidophilum 15%, lactobacillus delbruockii subspecies bulgaricus 15%, yeast saccharomyces cerevisiae 15%, bacillus megaterium 15%, Pasteur's Clostridium 15%.
The preparation method of compound microbial culture starter of the present invention, comprises the following steps:
One, inoculation, the temperature of the clean room specified by standard " Code for design of industrial clean rooms " GB50073-2001, humidity range and airblrne particulates cleanliness grade regulation, respectively by subtilis, Lactobacterium acidophilum, lactobacillus delbruockii subspecies bulgaricus, yeast saccharomyces cerevisiae, bacillus megaterium and Pasteur's Clostridium, inclined plane inoculating is carried out respectively by prior art, under room temperature (20 DEG C), relative humidity is 50%, cultivate 16-20h, then respectively join in sterilized water, vibrate and obtain bacterium liquid after 30 minutes, count with blood counting chamber, in bacterium liquid, bacterium quantity reaches 2-3 hundred million/ml respectively, preferably 200,000,000/ml.
Two, one-level cultivates (one-level spreads cultivation), being inoculated in respectively by the bacterial classification of described each bacterium liquid fills in the 50ml Erlenmeyer flask of level liquid substratum, inoculum size 5%-10% (v/v), preferably 10% (v/v), the pH value of substratum is 6.0-8.0, preferably 6.0-7.0, Erlenmeyer flask puts into shaking table with 180r/min, vibration rotating speed cultivates 24-48h, preferably 32h, obtains one-level and to spread cultivation bacterium liquid, count with blood counting chamber, in one-level spreads cultivation bacterium liquid, bacterium quantity reaches 2-3 hundred million/ml respectively, preferably 200,000,000/ml.
Level liquid substratum composition is respectively:
(1) subtilis level liquid substratum consists of: peptone 5.0g, beef leaching thing 3.0g, NaCl 5.0g, agar 15.0g, distilled water 1.0L.Mix rear 112 DEG C of sterilizing 20min, naturally cool to room temperature, pH7.0.Beef leaching thing (Beaf extract) adopts fresh beef through rejecting fat, digestion, filters, concentrated and a kind of brown color of obtaining is to tan paste, there is beef nature fragrance, soluble in water, the aqueous solution is faint yellow, also referred to as beef extract, extractum carnis, beef extract, beef powder, beef extract powder or beef extract powder.
(2) Lactobacterium acidophilum level liquid substratum consists of: casein peptone 10.0g, extractum carnis 10.0g, yeast powder 5.0g, glucose 5.0g, sodium acetate 5.0g, dibasic ammonium citrate 2.0g, sorbitan monooleate Soxylat A 25-7 (Tween 80) 1.0g, K
2hPO
42.0g, MgSO
4.7H
2o 0.2g, MnSO
4.H2O0.05g, CaCO
320.0g, agar 15.0g, distilled water 1.0L.Mix rear 112 DEG C of sterilizing 20min, naturally cool to 37 DEG C, pH6.8.
(3) lactobacillus delbruockii subspecies bulgaricus level liquid substratum consists of: casein peptone 10.0g, extractum carnis 10.0g, yeast powder 5.0g, glucose 5.0g, sodium acetate 5.0g, dibasic ammonium citrate 2.0g, Tween 801.0g, K
2hPO
42.0g, MgSO
4.7H
2o 0.2g, MnSO
4.H
2o 0.05g, CaCO
320.0g, agar 15.0g, distilled water 1.0L.Mix rear 112 DEG C of sterilizing 20min, naturally cool to 37 DEG C.pH6.8。
(4) yeast saccharomyces cerevisiae level liquid substratum consists of: 5 ° of B é wort 1.0L, agar 15.0g.Mix rear 112 DEG C of sterilizing 20min, natural in 28 DEG C, natural pH.
(5) bacillus megaterium level liquid substratum consists of: peptone 5.0g, beef leaching thing 3.0g, NaCl 5.0g, agar 15.0g, distilled water 1.0L.Mix rear 112 DEG C of sterilizing 20min, naturally cool to 30 DEG C, pH7.0.
(6) Pasteur's Clostridium level liquid substratum consists of: peptone 10.0g, beef extract 3.0g, NaCl 5.0g, agar 15.0g, distilled water 1.0L.Mix rear 112 DEG C of sterilizing 20min, naturally cool to culture temperature 37 DEG C, pH7.0.
One-level is cultivated and is act as the making bacterium liquid that spreads cultivation and cultivate for secondary.
Three, secondary cultivates (secondary spreads cultivation), each one-level being spread cultivation respectively to be inoculated into after bacterium liquid mixes with secondary liquid substratum is loaded with in the 1000ml Erlenmeyer flask of secondary liquid substratum, inoculum size 5%-10% (v/v), preferably 10% (v/v), Erlenmeyer flask is put into shaking table and is cultivated 48-72h with 180r/min vibration rotating speed, preferably 48h, obtain secondary to spread cultivation bacterium liquid, count with blood counting chamber, the secondary bacterium quantity in bacterium liquid that spreads cultivation reaches 2-3 hundred million/ml, preferably 200,000,000/ml respectively.
Secondary liquid substratum consists of: peptone 10.0g, extractum carnis 10.0g, agar 15.0g, molasses 5.0g, MgSO
4.7H
2o 0.2g, MnSO
4.H
2o 0.05g, CaCO
320.0g, NaCl 5.0g, distilled water 1.0L.Mix rear 112 DEG C of sterilizing 20min, naturally cool to room temperature, natural pH.
Secondary cultivate act as making spread cultivation bacterium liquid mixing after obtain composite bacteria liquid.
Four, by the spread cultivation mass percentage of bacterium liquid of each bacterial classification secondary be: subtilis 20-30%, Lactobacterium acidophilum 15-20%, lactobacillus delbruockii subspecies bulgaricus 15-20%, yeast saccharomyces cerevisiae 10-15%, bacillus megaterium 10-15%, Pasteur's Clostridium 10-15%, the mass percentage of good each bacterial classification is: subtilis 25%, Lactobacterium acidophilum 15%, lactobacillus delbruockii subspecies bulgaricus 15%, yeast saccharomyces cerevisiae 15%, bacillus megaterium 15%, Pasteur's Clostridium 15%, is mixed to get composite bacteria liquid.Composite bacteria liquid is adsorbed onto on granularity 200 object natural zeolite powder, granularity 200 object natural zeolite powder with have the mass ratio of the composite bacteria liquid of composite bacteria to be 1-3:1, then by prior art low-temperature vacuum drying, obtain compound microbial culture starter.
The bacterial classification that compound microbial culture starter of the present invention is used, for eliminating the foul smell that produces of organic waste and harmful pathogenic bacteria, comprehensive utilization organic waste also improves effect of fertilizer and quality and selects.Bacillus subtillis can produce subtilyne, nystatin and lactic acid isoreactivity antibacterial substance, and other pathogenic bacterium can be suppressed to grow; Produce a large amount of proteolytic enzyme, lipase, amylase and saccharifying enzyme, organic waste of degrading is as organic matter complicated in straw, changing food waste, mud simultaneously.Utilize yeast saccharomyces cerevisiae to produce acid and produce the feature of alcohol fragrance, organic waste can be reduced as the stink produced in the fermenting process such as manure of livestock and poultry, changing food waste.Lactobacillus delbruockii subspecies bulgaricus, Lactobacterium acidophilum can produce a large amount of lactic acid etc., can suppress harmful microorganism growth and breeding, can kill harmful bacteria as intestinal bacteria, Salmonellas, produce multivitamin and organic acid simultaneously, can promote plant growth.Bacillus megaterium has the effect of P and K decomposing, can supply Crop phosphoric and potassium element.Pasteur's Clostridium has nitrogen fixation, and supply nitrogen element is to Crop.
Embodiment 1
One, inoculate, by subtilis, Lactobacterium acidophilum, lactobacillus delbruockii subspecies bulgaricus, yeast saccharomyces cerevisiae, bacillus megaterium and Pasteur's Clostridium, inoculate respectively, under room temperature, humidity is 50%, cultivates 18h, then respectively joins in sterilized water, vibrate and obtain bacterium liquid after 30 minutes, in bacterium liquid, bacterium quantity reaches 200,000,000/ml respectively.
Two, one-level is cultivated, being inoculated in respectively by the bacterial classification of described each bacterium liquid fills in the 50ml Erlenmeyer flask of level liquid substratum, inoculum size 5%v/v, the pH value of substratum is 6.00, Erlenmeyer flask puts into shaking table with 180r/min shaking culture 32h, and in the bacterium liquid that one-level spreads cultivation, bacterium quantity reaches 200,000,000/ml respectively.
Three, secondary is cultivated, being inoculated into after each bacterium liquid one-level spread cultivation mixes with secondary liquid substratum is loaded with in the 1000ml Erlenmeyer flask of secondary liquid substratum, inoculum size 5%v/v, Erlenmeyer flask puts into shaking table with 180r/min shaking culture 48h, obtain secondary to spread cultivation bacterium liquid, the secondary bacterium quantity in bacterium liquid that spreads cultivation reaches 200,000,000/ml respectively.
Four, by the spread cultivation mass percentage of bacterium liquid of each bacterial classification secondary be: subtilis 25%, Lactobacterium acidophilum 20%, Lactobacillus delbrueckii kind 20%, yeast saccharomyces cerevisiae 15%, bacillus megaterium 10%, Pasteur's Clostridium 10%, is mixed to get composite bacteria liquid.Be adsorbed onto by the bacterium number of every for composite bacteria liquid ml of carrier 200,000,000 on granularity 200 order natural zeolite powder, the mass ratio of granularity 200 order natural zeolite powder and composite bacteria is 1:1, and low-temperature vacuum drying, obtains compound microbial culture starter.
Embodiment 2
One, inoculate, by subtilis, Lactobacterium acidophilum, lactobacillus delbruockii subspecies bulgaricus, yeast saccharomyces cerevisiae, bacillus megaterium and Pasteur's Clostridium, carry out inclined plane inoculating, under room temperature, humidity is 50%, cultivates 18h, then respectively joins in sterilized water, vibrate and obtain bacterium liquid after 30 minutes, in bacterium liquid, bacterium quantity reaches 200,000,000/ml respectively.
Two, one-level is cultivated, being inoculated in respectively by the bacterial classification of described each bacterium liquid fills in the 50ml Erlenmeyer flask of level liquid substratum, inoculum size 8%v/v, the pH value of substratum is 6.00, Erlenmeyer flask puts into shaking table with 180r/min shaking culture 32h, and in the bacterium liquid that one-level spreads cultivation, bacterium quantity reaches 200,000,000/ml respectively.
Three, secondary is cultivated, being inoculated into after each bacterium liquid one-level spread cultivation mixes with secondary liquid substratum is loaded with in the 1000ml Erlenmeyer flask of secondary liquid substratum, inoculum size 5%v/v, Erlenmeyer flask puts into shaking table with 180r/min shaking culture 48h, obtain secondary to spread cultivation bacterium liquid, the secondary bacterium quantity in bacterium liquid that spreads cultivation reaches 200,000,000/ml respectively.
Four, by the spread cultivation mass percentage of bacterium liquid of each bacterial classification secondary be: subtilis 20%, Lactobacterium acidophilum 15%, Lactobacillus delbrueckii kind 20%, yeast saccharomyces cerevisiae 15%, bacillus megaterium 15%, Pasteur's Clostridium 15%, is mixed to get composite bacteria liquid.Be adsorbed onto by the bacterium number of every for composite bacteria liquid ml of carrier 200,000,000 on granularity 200 order natural zeolite powder, the mass ratio of granularity 200 order natural zeolite powder and composite bacteria is 1:1, and low-temperature vacuum drying, obtains compound microbial culture starter.
Embodiment 3
One, inoculate, by subtilis, Lactobacterium acidophilum, lactobacillus delbruockii subspecies bulgaricus, yeast saccharomyces cerevisiae, bacillus megaterium and Pasteur's Clostridium, carry out inclined plane inoculating, under room temperature, humidity is 50%, cultivates 20h, then respectively joins in sterilized water, vibrate and obtain bacterium liquid after 30 minutes, in bacterium liquid, bacterium quantity reaches 300,000,000/ml respectively.
Two, one-level is cultivated, being inoculated in respectively by the bacterial classification of described each bacterium liquid fills in the 50ml Erlenmeyer flask of level liquid substratum, inoculum size 10%v/v, the pH value of substratum is 6.00, Erlenmeyer flask puts into shaking table with 180r/min shaking culture 32h, and in the bacterium liquid that one-level spreads cultivation, bacterium quantity reaches 300,000,000/ml respectively.
Three, secondary is cultivated, being inoculated into after each bacterium liquid one-level spread cultivation mixes with secondary liquid substratum is loaded with in the 1000ml Erlenmeyer flask of secondary liquid substratum, inoculum size 10%v/v, Erlenmeyer flask puts into shaking table with 180r/min shaking culture 48h, obtain secondary to spread cultivation bacterium liquid, the secondary bacterium quantity in bacterium liquid that spreads cultivation reaches 300,000,000/ml respectively.
Four, by the spread cultivation mass percentage of bacterium liquid of each bacterial classification secondary be: subtilis 20%, Lactobacterium acidophilum 20%, Lactobacillus delbrueckii kind 15%, yeast saccharomyces cerevisiae 15%, bacillus megaterium 15%, Pasteur's Clostridium 15%, is mixed to get composite bacteria liquid.Be adsorbed onto by the bacterium number of every for composite bacteria liquid ml of carrier 300,000,000 on granularity 200 order natural zeolite powder, the mass ratio of granularity 200 order natural zeolite powder and composite bacteria is 3:1, and low-temperature vacuum drying, obtains compound microbial culture starter.
Embodiment 4
One, inoculate, by subtilis, Lactobacterium acidophilum, lactobacillus delbruockii subspecies bulgaricus, yeast saccharomyces cerevisiae, bacillus megaterium and Pasteur's Clostridium, carry out inclined plane inoculating, under room temperature, humidity is 50%, cultivates 16h, then respectively joins in sterilized water, vibrate and obtain bacterium liquid after 30 minutes, in bacterium liquid, bacterium quantity reaches 200,000,000/ml respectively.
Two, one-level is cultivated, being inoculated in respectively by the bacterial classification of described each bacterium liquid fills in the 50ml Erlenmeyer flask of level liquid substratum, inoculum size 8%v/v, the pH value of substratum is 6.00, Erlenmeyer flask puts into shaking table with 180r/min shaking culture 24h, and in the bacterium liquid that one-level spreads cultivation, bacterium quantity reaches 200,000,000/ml respectively.
Three, secondary is cultivated, being inoculated into after each bacterium liquid one-level spread cultivation mixes with secondary liquid substratum is loaded with in the 1000ml Erlenmeyer flask of secondary liquid substratum, inoculum size 8%v/v, Erlenmeyer flask puts into shaking table with 180r/min shaking culture 72h, obtain secondary to spread cultivation bacterium liquid, the secondary bacterium quantity in bacterium liquid that spreads cultivation reaches 200,000,000/ml respectively.
Four, by the spread cultivation mass percentage of bacterium liquid of each bacterial classification secondary be: subtilis 30%, Lactobacterium acidophilum 15%, Lactobacillus delbrueckii kind 15%, yeast saccharomyces cerevisiae 15%, bacillus megaterium 10%, Pasteur's Clostridium 15%, is mixed to get composite bacteria liquid.Be adsorbed onto by the bacterium number of every for composite bacteria liquid ml of carrier 200,000,000 on granularity 200 order natural zeolite powder, the mass ratio of granularity 200 order natural zeolite powder and composite bacteria is 3:1, and low-temperature vacuum drying, obtains compound microbial culture starter.
Embodiment 5
One, inoculate, by subtilis, Lactobacterium acidophilum, lactobacillus delbruockii subspecies bulgaricus, yeast saccharomyces cerevisiae, bacillus megaterium and Pasteur's Clostridium, carry out inclined plane inoculating, under room temperature, humidity is 50%, cultivates 16h, then respectively joins in sterilized water, vibrate and obtain bacterium liquid after 30 minutes, in bacterium liquid, bacterium quantity reaches 200,000,000/ml respectively.
Two, one-level is cultivated, being inoculated in respectively by the bacterial classification of described each bacterium liquid fills in the 50ml Erlenmeyer flask of level liquid substratum, inoculum size 5%v/v, the pH value of substratum is 6.00, Erlenmeyer flask puts into shaking table with 180r/min shaking culture 48h, and in the bacterium liquid that one-level spreads cultivation, bacterium quantity reaches 200,000,000/ml respectively.
Three, secondary is cultivated, being inoculated into after each bacterium liquid one-level spread cultivation mixes with secondary liquid substratum is loaded with in the 1000ml Erlenmeyer flask of secondary liquid substratum, inoculum size 5%v/v, Erlenmeyer flask puts into shaking table with 180r/min shaking culture 72h, obtain secondary to spread cultivation bacterium liquid, the secondary bacterium quantity in bacterium liquid that spreads cultivation reaches 200,000,000/ml respectively.
Four, by the spread cultivation mass percentage of bacterium liquid of each bacterial classification secondary be: subtilis 30%, Lactobacterium acidophilum 20%, Lactobacillus delbrueckii kind 15%, yeast saccharomyces cerevisiae 10%, bacillus megaterium 15%, Pasteur's Clostridium 10%, is mixed to get composite bacteria liquid.Be adsorbed onto by the bacterium number of every for composite bacteria liquid ml of carrier 200,000,000 on granularity 200 order natural zeolite powder, the mass ratio of granularity 200 order natural zeolite powder and composite bacteria is 1:1, and low-temperature vacuum drying, obtains compound microbial culture starter.
Application examples 1
Adopt the compound microbial culture starter of embodiment 1, fermentation maturity major ingredient A, major ingredient B and auxiliary material, make powdery bio-organic fertilizer.The raw material of major ingredient A is the wet chicken manure that quality solid content is no less than 70%, and the raw material of major ingredient B is the sewage sludge that quality solid content is no less than 70%, and the raw material of auxiliary material is dry straw.
Preparation method is as follows:
One, rice straw powder is broken to no longer than 10mm, makes auxiliary material.
Two, will wet chicken manure 700 kilograms, auxiliary material 300 kilograms and compound microbial culture starter 2 kilograms, mix, pile high 1-2 rice, turning 1 time after 15 days, stack and become thoroughly decomposed 30 days, make major ingredient A.
Domestic sewage sludge 700 kilograms, auxiliary material 300 kilograms and compound microbial culture starter 2 is public, mix, pile high 1-2 rice, turning 1 time after 15 days, stack and become thoroughly decomposed 30 days, make major ingredient B.
Three, by major ingredient A500 kilogram, major ingredient B500 kilogram, the mixing of amine sulfate 50 kilograms, calcium superphosphate 50 kilograms, 50 kilograms, magnesium aluminum silicate powder is even ferments all afterwards, during fermentation keep temperature 45-65 DEG C, when leavening temperature is more than 65 DEG C, adopt stack turner turning cooling, lump with anti-fermentative material and make it mix, stacking is become thoroughly decomposed 25 days, completes fermentation.
Four, will the crushing material of fermentation be completed, and make the compost finished product that lumpd broken, then sieve and remove impurity further.
Five, be metered to bag packaging, obtain powdery biological organic fertilizer.
By NY884-2012 " biological organic fertilizer " standard testing, its active principle is organic >=40%, living bacteria count amount cfu >=0.2 hundred million/gram, coliform group count≤100/g (mL), induced worm egg death rate >=95%, water ratio≤30%, without foul smell.
Application examples 2
Adopt the compound microbial culture starter of embodiment 2, fermentation maturity major ingredient A, major ingredient B and auxiliary material, make powdery bio-organic fertilizer.The raw material of major ingredient A is the wet pig manure that quality solid content is no less than 70%, and the raw material of major ingredient B is the wet chicken manure that quality solid content is no less than 70%, and the raw material of auxiliary material is the mushroom slag that mass water content is not more than 35%.
Preparation method is as follows:
One, mushroom ground-slag is broken to is not more than 10mm size, make auxiliary material.
Two, will wet pig manure 700 kilograms, auxiliary material 300 kilograms and compound microbial culture starter 2 kilograms, mix, pile high 1-2 rice, turning 1 time after 15 days, stack and become thoroughly decomposed 30 days, make major ingredient A.
To be wet chicken manure 700 kilograms, auxiliary material 300 kilograms and compound microbial culture starter 2 kilograms, and uniformly dispersing and mixing are piled between high 1-2 rice, turning 1 time after 15 days, stack and become thoroughly decomposed 30 days, make major ingredient B.
Three, by major ingredient A500 kilogram, major ingredient B500 kilogram, amine sulfate 50 kilograms, calcium superphosphate 50 kilograms, 50 kilograms, magnesium aluminum silicate powder, mix and evenly to ferment all afterwards, during fermentation keep temperature 45-65 DEG C, when leavening temperature is more than 65 DEG C, adopt stack turner turning cooling, lump with anti-fermentative material and make it mix, stacking and become thoroughly decomposed 15 days.
Four, will the crushing material of fermentation be completed, and make the compost finished product that lumpd broken, then sieve and remove impurity further.
Five, be metered to bag packaging, obtain powdery biological organic fertilizer.
By NY884-2012 " biological organic fertilizer " standard testing, its active principle is organic >=40%, living bacteria count amount cfu >=0.2 hundred million/gram, excrement colibacillus group number≤100/g (mL), induced worm egg death rate >=95%, water ratio≤30%, without foul smell.
Application examples 3
Adopt the compound microbial culture starter of embodiment 3, fermentation maturity major ingredient A, major ingredient B and auxiliary material, make powdery bio-organic fertilizer.The raw material of major ingredient A is the cow dung that quality solid content is no less than 70%, and the raw material of major ingredient B is the sheep excrement that quality solid content is no less than 70%, and the raw material of auxiliary material is dry cotton stalk.
Preparation method is as follows:
One, by cotton stalk crushing to being not more than 10mm size, make auxiliary material.
Two, by cow dung 700 kilograms, auxiliary material 300 kilograms and compound microbial culture starter 2 kilograms, mix, pile high 1-2 rice, turning 1 time after 15 days, stack and become thoroughly decomposed 30 days, make major ingredient A.
By 700 kilograms, sheep excrement, auxiliary material 300 kilograms and compound microbial culture starter 2 kilograms, mix, pile high 1-2 rice, turning 1 time after 15 days, stack and become thoroughly decomposed 30 days, make major ingredient B.
Three, by major ingredient A500 kilogram, major ingredient B500 kilogram, amine sulfate 50 kilograms, calcium superphosphate 50 kilograms, 50 kilograms, magnesium aluminum silicate powder, mix and evenly to ferment all afterwards, during fermentation keep temperature 45-65 DEG C, when leavening temperature is more than 65 DEG C, adopt stack turner turning cooling, lump with anti-fermentative material and make it mix, stacking and become thoroughly decomposed 15 days.
Four, will the crushing material of fermentation be completed, and make the compost finished product that lumpd broken, then sieve and remove impurity further.
Five, be metered to bag packaging, obtain powdery biological organic fertilizer.
By NY884-2012 " biological organic fertilizer " standard testing, its active principle is organic >=40%, living bacteria count amount cfu >=0.2 hundred million/gram, excrement colibacillus group number≤100/g (mL), induced worm egg death rate >=95%, water ratio≤30%, without foul smell.
Claims (10)
1. a compound microbial culture starter, it is characterized in that: described compound microbial culture starter is made up of carrier and composite bacteria, mix with the mass ratio 1-3:1 of the composite bacteria liquid of composite bacteria by carrier, obtain after drying, described carrier is granularity 200 object natural zeolite powder, described composite bacteria liquid is made up of the mass percent of following bacterial classification bacterium liquid: subtilis 20-30%, Lactobacterium acidophilum 15-20%, lactobacillus delbruockii subspecies bulgaricus 15-20%, yeast saccharomyces cerevisiae 10-15%, bacillus megaterium 10-15%, Pasteur's Clostridium 10-15%, described each bacterial classification bacterium liquid every ml of carrier 2-3 hundred million bacterium number.
2. compound microbial culture starter according to claim 1, it is characterized in that: described composite bacteria liquid is made up of the mass percent of following bacterium liquid: subtilis 25%, Lactobacterium acidophilum 15%, lactobacillus delbruockii subspecies bulgaricus 15%, yeast saccharomyces cerevisiae 15%, bacillus megaterium 15%, Pasteur's Clostridium 15%, described each bacterium liquid every ml of carrier 200,000,000 bacterium number.
3. a preparation method for compound microbial culture starter, comprises the following steps:
One, inoculate, respectively by subtilis, Lactobacterium acidophilum, lactobacillus delbruockii subspecies bulgaricus, yeast saccharomyces cerevisiae, bacillus megaterium and Pasteur's Clostridium, under room temperature, relative humidity is 50%, cultivate 16-20h, then respectively join in sterilized water, vibrate and obtain bacterium liquid after 30 minutes;
Two, one-level is cultivated, described each bacterium liquid is inoculated in the Erlenmeyer flask filling level liquid substratum respectively, inoculum size is 5%-10%, the pH value of substratum is 6.0-8.0,24-48h is cultivated with 180r/min rotating speed, obtain one-level to spread cultivation bacterium liquid, the one-level bacterium quantity in bacterium liquid that spreads cultivation reaches 2-3 hundred million/ml respectively;
Three, secondary is cultivated, each one-level being spread cultivation after bacterium liquid mixes with secondary liquid substratum is inoculated in the Erlenmeyer flask being loaded with secondary liquid substratum, inoculum size is 5%-10%, 48-72h is cultivated with 180r/min rotating speed, obtain secondary to spread cultivation bacterium liquid, the secondary bacterium quantity in bacterium liquid that spreads cultivation reaches 2-3 hundred million/ml respectively;
Four, by the spread cultivation mass percentage of bacterium liquid of each bacterial classification secondary be: subtilis 20-30%, Lactobacterium acidophilum 15-20%, lactobacillus delbruockii subspecies bulgaricus 15-20%, yeast saccharomyces cerevisiae 10-15%, bacillus megaterium 10-15%, Pasteur's Clostridium 10-15%, is mixed to get composite bacteria liquid; Composite bacteria liquid is adsorbed onto on granularity 200 object natural zeolite powder, granularity 200 object natural zeolite powder with have the mass ratio of the composite bacteria liquid of composite bacteria to be 1-3:1, dry, obtain compound microbial culture starter.
4. the preparation method of compound microbial culture starter according to claim 3, is characterized in that: described step 2 inoculum size is 10%, and the pH value of described substratum is 6.0-7.0; Described with 180r/min rotating speed, cultivate 32h.
5. the preparation method of compound microbial culture starter according to claim 4, is characterized in that: bacterium quantity 200,000,000/ml respectively in described step 2 bacterium liquid.
6. the preparation method of compound microbial culture starter according to claim 5, is characterized in that: the level liquid substratum composition of described step 2 is respectively:
(1) subtilis level liquid substratum consists of: peptone 5.0g, beef leaching thing 3.0g, NaCl 5.0g, agar 15.0g, distilled water 1.0L; Mix rear 112 DEG C of sterilizing 20min, naturally cool to room temperature, pH7.0;
(2) Lactobacterium acidophilum level liquid substratum consists of: casein peptone 10.0g, extractum carnis 10.0g, yeast powder 5.0g, glucose 5.0g, sodium acetate 5.0g, dibasic ammonium citrate 2.0g, sorbitan monooleate Soxylat A 25-7 1.0g, K
2hPO
42.0g, MgSO
4.7H
2o 0.2g, MnSO
4.H2O 0.05g, CaCO
320.0g, agar 15.0g, distilled water 1.0L; Mix rear 112 DEG C of sterilizing 20min, naturally cool to 37 DEG C, pH6.8;
(3) lactobacillus delbruockii subspecies bulgaricus level liquid substratum consists of: casein peptone 10.0g, extractum carnis 10.0g, yeast powder 5.0g, glucose 5.0g, sodium acetate 5.0g, dibasic ammonium citrate 2.0g, Tween 801.0g, K
2hPO
42.0g, MgSO
4.7H
2o 0.2g, MnSO
4.H
2o 0.05g, CaCO
320.0g, agar 15.0g, distilled water 1.0L; Mix rear 112 DEG C of sterilizing 20min, naturally cool to 37 DEG C, pH6.8;
(4) yeast saccharomyces cerevisiae level liquid substratum consists of: 5 ° of B é wort 1.0L, agar 15.0g; Mix rear 112 DEG C of sterilizing 20min, natural in 28 DEG C, natural pH;
(5) bacillus megaterium level liquid substratum consists of: peptone 5.0g, beef leaching thing 3.0g, NaCl 5.0g, agar 15.0g, distilled water 1.0L; Mix rear 112 DEG C of sterilizing 20min, naturally cool to 30 DEG C, pH7.0;
(6) Pasteur's Clostridium level liquid substratum consists of: peptone 10.0g, beef extract 3.0g, NaCl 5.0g, agar 15.0g, distilled water 1.0L; Mix rear 112 DEG C of sterilizing 20min, naturally cool to culture temperature 37 DEG C, pH7.0.
7. the preparation method of compound microbial culture starter according to claim 6, is characterized in that: described step 3 inoculum size is 10%; Described with 180r/min rotating speed cultivation 48h.
8. the preparation method of compound microbial culture starter according to claim 7, is characterized in that: the secondary of the described step 3 bacterium quantity in bacterium liquid that spreads cultivation is respectively 200,000,000/ml.
9. the preparation method of compound microbial culture starter according to claim 8, is characterized in that: the secondary liquid substratum of described step 3 consists of: peptone 10.0g, extractum carnis 10.0g, agar 15.0g, molasses 5.0g, MgSO
4.7H
2o 0.2g, MnSO
4.H
2o 0.05g, CaCO
320.0g, NaCl 5.0g, distilled water 1.0L; Mix rear 112 DEG C of sterilizing 20min, naturally cool to room temperature, natural pH.
10. the preparation method of compound microbial culture starter according to claim 9, it is characterized in that: the spread cultivation mass percentage of bacterium liquid of each bacterial classification secondary of described step 4 is: subtilis 25%, Lactobacterium acidophilum 15%, lactobacillus delbruockii subspecies bulgaricus 15%, yeast saccharomyces cerevisiae 15%, bacillus megaterium 15%, Pasteur's Clostridium 15%.
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| CN106220268A (en) * | 2016-07-12 | 2016-12-14 | 德清龙溪农业科技有限公司 | A kind of for Cucurbita pepo L. organic fertilizer |
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| CN108795812A (en) * | 2018-06-25 | 2018-11-13 | 湖南泰谷生态工程有限公司 | A kind of compost decomposing agent and the preparation method and application thereof |
| CN110699349A (en) * | 2019-10-28 | 2020-01-17 | 广西科技师范学院 | Microbial food leavening agent and preparation method thereof |
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| CN111748495A (en) * | 2020-07-02 | 2020-10-09 | 农业农村部规划设计研究院 | Initiating agent for cow dung aerobic fermentation and preparation method thereof |
| CN112063557A (en) * | 2020-09-17 | 2020-12-11 | 杭州楠大环保科技有限公司 | Perishable garbage low-temperature phase change water-producing degradation microbial inoculum and microbial liquid preparation method |
| CN114380629A (en) * | 2022-02-17 | 2022-04-22 | 重庆市农业科学院 | Composite microbial straw decomposing inoculant and preparation method thereof |
| CN115057730A (en) * | 2022-04-01 | 2022-09-16 | 北京四良科技有限公司 | Fermentation process method of low-emission micro-oxygen fermentation machine |
| CN115057730B (en) * | 2022-04-01 | 2023-05-05 | 北京四良科技有限公司 | Fermentation process method of low-emission micro-oxygen fermentation machine |
| CN115160081A (en) * | 2022-06-16 | 2022-10-11 | 西安智同天地环境工程有限公司 | Composting method suitable for low-temperature environment |
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