CN102286376B - Microbial inoculum for high-efficiency fermenting bed and preparation method thereof - Google Patents
Microbial inoculum for high-efficiency fermenting bed and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a microbial inoculum for a high-efficiency fermenting bed and a preparation method thereof. The microbial inoculum is prepared by thoroughly and evenly mixing 30-40% of Bacillus subtilis leavening powder, 30-40% of Bacillus licheniformis leavening powder and 20-30% of Candida utilis leavening powder. The microbial inoculum can effectively promote in-situ degradation of livestock and poultry excrement, obviously reduce the generation of ammonia and other putrilages in the livestock and poultry excrement, and enhance the quality of pork.
Description
Technical field
The present invention relates to the microbial fermentation field, definite say that a kind of fermentation bed is with microbiobacterial agent and preparation method thereof.
Background technology
Along with China's pig industry mass-producing, intensive fast development, feces of livestock and poultry and sewage discharge are aobvious serious to the pollution day of environment in recent years.According to statistics, the solid waste total amount that now national annual herding is produced is about 3,000,000,000 tons, only Changsha annual just approximately has 8,000 ten thousand tons of livestock breeding wastewaters, more than 1,200,000 tons, solid manure is discharged, most of plant has caused larger pollution only through just directly effluxing after the methane-generating pit simple process to environment.
The fecaluria treatment technology that present tradition is raised pigs adopts the modes such as compost fermentation, biogas fermentation, aerobic fermentation drying technology to process mostly, compost fermentation is processed the ight soil land occupation, atmospheric pollution to environment is larger, biogas fermentation has the limitation in season, aerobic fermentation drying technology cost is higher, and certain limitation is all arranged in actual utilization.
Probiotics fermention bed to raise pig technology is theoretical according to little Ecological Principle and biological fermentation, agricultural crop straw, microbiobacterial agent and the assisted fermentation agent such as rice bran, sugar of saw dust, rice husk, pulverizing are mixed by a certain percentage, be layered in fermenting tank for hog house, spontaneous fermentation, form microbial fermentation bed, feeding live pig utilizes microorganism to livestock and poultry fecaluria " original position degraded " on it, reaches the Novel cultivation pattern of ecotope " zero pollutes ".
Because proportioning, thickness, the residing Geographical environment of fermenting bed padding are different, make ventilation property, water-absorbent, atmospheric moisture and the envrionment temperature of bedding and padding different; , must degrade timely constantly producing movement due to live pig, therefore select safety, stable, colonization ability is strong, degradation rate is high and function to have the complex micro organism fungicide that the bacterial strain of complementary action forms be the favourable assurance of fermentation bed cleaning cultivation.
Summary of the invention
Purpose of the present invention aims to provide a kind of effective promotion livestock and poultry fecaluria " original position degraded ", significantly reduces the generation of ammonia and other septic matter in the livestock and poultry fecaluria, and the fermentation bed that can improve meat quality is with microbiobacterial agent and preparation method thereof.
The objective of the invention is to realize in the following manner:
A kind of high-efficiency fermenting bed preparation method of complex micro organism fungicide comprises the following steps:
1) fermentation of bacillus subtilis thing preparation
At first, subtilis is cultivated through slant strains activation, shake-flask seed;
Then carry out fermentor cultivation: in fermentor tank, the substratum loading amount is 50～70% of cumulative volume, inoculum size accounts for 0.1～0.3% of culture volume, 34～37 ℃ of leavening temperatures, blowing air 180～200rpm stir culture, the ratio of ventilation volume and fermentating liquid volume 0.9: 1～1: 1, tank pressure 0.03～0.05MPa; Fermentation termination be the gemma number be not less than total count 90%, and the viable count of fermented liquid is 1.0 * 10
10～1.5 * 10
10Between cfu/mL; Each constituent mass per-cent of fermentation tank culture medium is: glucose 0.5～0.7%, starch 0.3～0.5%, soybean cake powder 0.8～1.2%, manganous sulfate 0.0062%, yeast powder 0.5～0.8%, peptone 0.3～0.5%, iron(ic) chloride 0.01%, potassium primary phosphate 0.4%, calcium carbonate 0.1%, sal epsom 0.05%, all the other are water, pH 7.2～7.4;
Carry out at last solid absorption: fermented liquid adsorbs rear low temperature dehumidifier drying with the part by weight absorption of wheat bran by 1: 2～1: 3, pulverizes, and the powder product bacteria containing amount is 2.0 * 10
9～5.0 * 10
9Between cfu/g;
2) the lichen bacillus ferments thing preparation
At first, Bacillus licheniformis is cultivated through slant strains activation, shake-flask seed; Described Bacillus licheniformis, preserving number are CGMCC No.4835;
Then carry out fermentor cultivation: in fermentor tank, the substratum loading amount is 50～70% of cumulative volume, inoculum size accounts for 0.1～0.3% of culture volume, 45～50 ℃ of leavening temperatures, blowing air 180～200rpm stir culture, the ratio of ventilation volume and fermentating liquid volume 0.9: 1～1: 1, tank pressure remains on 0.03～0.05MPa, fermentation termination be the gemma number be not less than total count 90%, and fermented liquid contain the bacterium number 8.0 * 10
9～1.2 * 10
10Between cfu/mL; Each constituent mass per-cent of fermentation tank culture medium is: starch 1.0～1.5%, dextrin 0.5～0.8%, soybean cake powder 1.2～1.5%, ammonium sulfate 0.6～0.8%, yeast powder 0.3～0.4%, calcium carbonate 0.2%, sodium-chlor 0.4%, potassium primary phosphate 0.02%, all the other are water, pH 7.0～7.2;
Carry out at last solid absorption: fermented liquid adsorbs rear low temperature dehumidifier drying with the part by weight absorption of wheat bran by 1: 2～1: 3, pulverizes, and the powder product bacteria containing amount is 1.5 * 10
9～4.0 * 10
9Between cfu/g;
3) Candida utilis fermented product preparation
At first, Candida utilis is cultivated through slant strains activation, shake-flask seed;
Then carry out fermentor cultivation: in fermentor tank, the substratum loading amount is 50～70% of cumulative volume, inoculum size accounts for 0.3～0.5% of culture volume, 25～28 ℃ of leavening temperatures, blowing air 120～150rpm stir culture, the ratio of ventilation volume and fermentating liquid volume 1: 1～1.1: 1, tank pressure remains on 0.03～0.05MPa, 46～48 hours complete secondary fermentation liquid that ferments contain the bacterium number 2.0 * 10
9～3.0 * 10
9Between cfu/mL; Each constituent mass per-cent of fermentation tank culture medium is: glucose 3.5～4.0%, and ammonium sulfate 1.5～2.0%, yeast powder 0.2～0.25%, potassium primary phosphate 0.1%, sodium-chlor 0.01%, sal epsom 0.05%, all the other are water, pH 6.0～6.5;
Carry out at last solid absorption: fermented liquid adsorbs rear low temperature dehumidifier drying with the part by weight absorption of wheat bran by 1: 1, pulverizes, and the powder product bacteria containing amount is 1.5 * 10
9～2.5 * 10
9Between cfu/g;
4) high-efficiency fermenting bed is composite with complex micro organism fungicide
Careless fermentation of bacillus medicinal powder 30～40% by weight percentage, the lichen bacillus ferments medicinal powder 30～40%, Candida utilis fermented product pulvis 20%～30%, fully mixing gets final product.
Described high-efficiency fermenting bed is not less than 1.0 * 10 with the total viable count of complex micro organism fungicide
9Cfu/g.
Step 1) each constituent mass per-cent of substratum of described slant strains activation is: peptone 1%, and NaCl 0.5%, yeast extract paste 0.5%, agar 1.5%～2.0%, all the other are water, pH 7.2-7.4;
Step 1) substratum cultivated of described shake-flask seed is in step 1) do not add agar in described slant medium, concrete operations: fresh slant strains one ring of getting activation, be inoculated in shake-flask seed liquid substratum, loading amount is 100mL/500mL, rotating speed is 180～200rpm, after 34～37 ℃ of constant temperature culture 22～24h, 80 ℃ of water-bath thermal treatment 10 minutes gets final product.
Step 2) each constituent mass per-cent of substratum of described slant strains activation is: glucose 0.4%, and malt extract 1%, yeast extract 0.4%, calcium carbonate 0.2%, agar 1.5%～2.0%, all the other are water, pH 7.0～7.2;
Step 2) substratum cultivated of described shake-flask seed is in step 2) do not add agar in described slant medium, concrete operations: fresh slant strains one ring of getting activation, be inoculated in shake-flask seed liquid substratum, loading amount is 100mL/500mL, rotating speed is 180～200rpm, after 45～50 ℃ of constant temperature culture 28～30h, stop cultivating.
Step 3) substratum of described slant strains activation is the PDA substratum;
Step 3) described shake-flask seed is cultivated concrete operations: fresh slant strains one ring of getting activation, be inoculated in shake-flask seed liquid substratum, loading amount is 150mL/500mL, and rotating speed is 120～150rpm, after 25～28 ℃ of constant temperature culture 30～32h, stop cultivating; Each constituent mass per-cent of the substratum of described shake-flask seed: glucose 2%, peptone 2%, yeast extract 1%, all the other are water, pH 6.0～6.5.
A kind of microbial inoculum for high-efficiency fermenting bed is the microbial inoculum that is prepared from by above-mentioned method.Described Bacillus licheniformis, preserving number are CGMCC No.4835.
Advantage of the present invention:
The preponderant strains selection of species
Subtilis: aerobic bacteria, can produce proteolytic enzyme, the multiple enzyme such as amylase can obviously improve growth of animal speed and efficiency of feed utilization.Higher stability is arranged in the feed course of processing and sour environment, and reproduction speed is fast, good stress resistance, and decomposition fecaluria ability is strong.Be 30 ℃～37 ℃ through the bacterial strain optimal temperature after optimizing, be inoculated in liquid fermentation medium, produce proteolytic enzyme during 24h and be respectively 2706u/mL, 31.331MMU with the product amylase activity; Be seeded in pig manure urine extracting solution and react after 25 days, to degradation rate, the NH of COD
3-N degradation rate is respectively 86.5%, 70.7%.
Bacillus licheniformis: screening and separating obtains from the pig manure that becomes thoroughly decomposed of health pig discharging, the high growth temperature bacterium.Can produce proteolytic enzyme, amylase, cellulase etc.At hot stage, bacterium is more active.Be 45 ℃～55 ℃ through the bacterial strain optimal temperature after optimizing, be seeded in pig manure urine extracting solution and react after 25 days, to NH
3-N degradation rate is 75.2%.
Candida utilis: the oxygen bacterium of holding concurrently, 25 ℃～28 ℃ of suitable growth temperatures, pH value 6.0～6.5.For animal provides protein and VITAMIN, help digest, the stimulating animal growth suppresses the sex pheromone breeding, improves immunity of organisms.Be seeded in pig manure urine extracting solution and react after 2 days, to NH
3-N degradation rate is 83.5%.
Product fermentation bed of the present invention with complex micro organism fungicide be seeded in pig manure urine extracting solution reaction after 25 days the COD degradation rate reached 89.9%, NH
3-N clearance reaches 84.5%.
The preservation information of bacterial strain of the present invention is as follows:
Preservation date: on May 9th, 2011
Depositary institution's title: China Committee for Culture Collection of Microorganisms's common micro-organisms center (being called for short CGMCC)
Deposit number: CGMCC No.4835
Classification And Nomenclature: Bacillus licheniformis (Bacillus licheniformis).
Beneficial effect of the present invention
Product of the present invention is according to the having complementary functions property structure of the useful bacterial strain of difference, and it forms clear and definite, steady quality.After particularly passing through the unique zymotechnique of the present invention, what in fact obtain is the mixture of microbial inoculum and fermentating metabolism product, may can produce many useful materials.By the production testing discovery of reality, product of the present invention is used for fermentation bed to raise pig and can utilizes microorganism to livestock and poultry fecaluria " original position degraded ", reaches and reduces the pig farm to the pollution of ecotope, realizes the recycling ecological agriculture purpose; Simultaneously can improve the live pig welfare, improve meat quality.
Embodiment
Following embodiment is intended to further illustrate the present invention, and unrestricted the present invention.
The 3 strain strain name that the present invention adopts and bacterial classification source are as follows:
Subtilis (ACCC10619) is bought in Chinese agriculture microbial strains preservation center;
Bacillus licheniformis (Bacillus licheniformis), screening and separating obtains from the pig manure that becomes thoroughly decomposed of health pig discharging, identifies through institute of microbiology of the Chinese Academy of Sciences.And carried out preservation on May 9th, 2011 at China Committee for Culture Collection of Microorganisms's common micro-organisms center, and Bacillus licheniformis (Bacillus licheniformis), preserving number is CGMCC No.4835.
Screening process is as follows: the healthy pig manure sample that will become thoroughly decomposed and enrichment medium (fresh pig manure) (weight ratio) by a certain percentage fully mix, progressively add wood sawdust and rice husk, regulate the water ratio of material in 55% left and right, be loaded in clean plastic tank, static incubated at room temperature, stir every day for several times, measure foul smell variation and weight-loss ratio changing conditions in culturing process, until the basic odorless of fecaluria and loss of weight are when not obvious, culture transferring continues shaking culture, progressively improve enrichment medium concentration to 95%, each is taken turns and is cultured to odorless, when loss of weight is not obvious till.The enrichment culture thing by repeatedly dull and stereotyped coating separation, is obtained strong bacterial strain 4 strains of fecaluria capacity of decomposition.Wherein 1 strain bacterial strain bacterium colony is larger, and the edge is irregular; It is shaft-like that thalline is, and gemma is arranged, Gram-positive; Starch Hydrolysis test, gelatin hydrolysis, oxidase test, catalase test, methyl red test, citrate test, VP＜pH6 test positive; VP＞pH7 negative.Be accredited as Bacillus licheniformis through institute of microbiology of the Chinese Academy of Sciences.
Candida utilis (ACCC20060) is bought in Chinese agriculture microbial strains preservation center.
Embodiment 1: the composite fungus agent preparation
1. fermentation of bacillus subtilis thing preparation
The slant strains activation: get the Refrigerator store slant strains, aseptic technique is inoculated in fresh test tube slant substratum, and 37 ℃ of constant temperature culture are spent the night, and make fresh test tube slant bacterial classification.Slant medium: peptone 1%, NaCl 0.5%, yeast extract paste 0.5%, agar 1.5%～2.0%, all the other are water, pH 7.2-7.4.
Shake-flask seed is cultivated: get fresh slant strains one ring of activation, aseptic technique is inoculated in shake-flask seed liquid substratum, and loading amount is 100mL/500mL, rotating speed is 180～200rpm, after 34～37 ℃ of constant temperature culture 22～24h, 80 ℃ of water-bath thermal treatment 10 minutes gets final product.The substratum of described shake-flask seed is not add agar in slant medium.
Fermentor cultivation: in fermentor tank, the substratum loading amount is 50～70% of cumulative volume, inoculum size accounts for 0.1～0.3% of culture volume, 34～37 ℃ of leavening temperatures, blowing air stir culture (180～200rpm), ventilation volume was than 0.9: 1～1: 1, tank pressure remains on 0.03-0.05MPa, suitably adjusts free air delivery and tank pressure according to the foam situation after inoculation, prevents from escaping liquid.After foam fades away, strengthen air flow.Every 4 hours sampling microscopies once.Fermentation termination is that the gemma number accounts for more than 90% of total count.Put the tank sample and measure total count with colony counting method.Fermented liquid contain the bacterium number 1.0 * 10
10-1.5 * 10
10Between cfu/ml.Fermentation tank culture medium: glucose 0.5～0.7%, starch 0.3～0.5%, soybean cake powder 0.8～1.2%, manganous sulfate 0.0062%, yeast powder 0.5～0.8%, peptone 0.3～0.5%, iron(ic) chloride 0.01%, potassium primary phosphate 0.4%, calcium carbonate 0.1%, sal epsom 0.05%, all the other are water, pH 7.2～7.4.
Solid absorption: fermented liquid adsorbs rear low temperature dehumidifier drying with the part by weight absorption of wheat bran by 1: 2～1: 3, pulverizes, and after measuring bacteria containing amount, puts preservation in storage.The powder product bacteria containing amount is 2.0 * 10
9-5.0 * 10
9Between cfu/g.
2. the lichen bacillus ferments thing preparation
Each constituent mass per-cent of substratum of slant strains activation is: glucose 0.4%, and malt extract 1%, yeast extract 0.4%, calcium carbonate 0.2%, agar 1.5%～2.0%, all the other are water, pH 7.0～7.2;
Get fresh slant strains one ring of activation, be inoculated in shake-flask seed liquid substratum, loading amount is 100mL/500mL, and rotating speed is 180～200rpm, after 45～50 ℃ of constant temperature culture 28～30h, stops cultivating.The substratum of described shake-flask seed is not add agar in slant medium.
Then carry out fermentor cultivation: in fermentor tank, the substratum loading amount is 50～70% of cumulative volume, inoculum size accounts for 0.1～0.3% of culture volume, 45～50 ℃ of leavening temperatures, blowing air stir culture (180～200rpm), ventilation volume was than (ratio of air flow and fermentating liquid volume) 0.9: 1～1: 1, tank pressure remains on 0.03～0.05MPa, fermentation termination be the gemma number be not less than total count 90%; Fermented liquid contain the bacterium number 8.0 * 10
9～1.2 * 10
10Between cfu/mL; Each constituent mass per-cent of fermentation tank culture medium is: starch 1.0～1.5%, dextrin 0.5～0.8%, soybean cake powder 1.2～1.5%, ammonium sulfate 0.6～0.8%, yeast powder 0.3～0.4%, calcium carbonate 0.2%, sodium-chlor 0.4%, potassium primary phosphate 0.02%, all the other are water, pH 7.0～7.2;
Carry out at last solid absorption: fermented liquid adsorbs rear low temperature dehumidifier drying with the part by weight absorption of wheat bran by 1: 2～1: 3, pulverizes, and the powder product bacteria containing amount is 1.5 * 10
9～4.0 * 10
9Between cfu/g.
3. Candida utilis fermented product preparation
The substratum of slant strains activation is the PDA substratum;
Get fresh slant strains one ring of activation, be inoculated in shake-flask seed liquid substratum, loading amount is 150mL/500mL, and rotating speed is 120～150rpm, after 25～28 ℃ of constant temperature culture 30～32h, stops cultivating.Each constituent mass per-cent of the substratum of described shake-flask seed: glucose 2%, peptone 2%, yeast extract 1%, all the other are water, pH 6.0～6.5.
Then carry out fermentor cultivation: in fermentor tank, the substratum loading amount is 50～70% of cumulative volume, inoculum size accounts for 0.3～0.5% of culture volume, 25～28 ℃ of leavening temperatures, blowing air stir culture (120～150rpm), ventilation volume was than (ratio of air flow and fermentating liquid volume) 1: 1～1.1: 1, tank pressure remains on 0.03～0.05MPa, and fermentation in 46～48 hours is complete; Fermented liquid contain the bacterium number 2.0 * 10
9～3.0 * 10
9Between cfu/mL; Each constituent mass per-cent of fermentation tank culture medium is: glucose 3.5～4.0%, and ammonium sulfate 1.5～2.0%, yeast powder 0.2～0.25%, potassium primary phosphate 0.1%, sodium-chlor 0.01%, sal epsom 0.05%, all the other are water, pH 6.0～6.5;
Carry out at last solid absorption: fermented liquid adsorbs rear low temperature dehumidifier drying with the part by weight absorption of wheat bran by 1: 1, pulverizes, and the powder product bacteria containing amount is 1.5 * 10
9～2.5 * 10
9Between cfu/g.
4. zymophyte is composite
The fermentation of bacillus subtilis medicinal powder 30～40% by weight percentage, the lichen bacillus ferments medicinal powder 30～40%, and Candida utilis fermented product pulvis 20～30%, fully mixing final vacuum packing namely obtains composite fungus agent.
Embodiment 2: utilize composite fungus agent of the present invention to prepare the effect of raising pigs of fermentation bed
1. fermentation bed preparation
Fermenting bed padding is made of by the weight ratio of 1: 1 saw dust, rice husk, or is made of by the weight ratio of 5: 3: 2 saw dust, husk, agricultural crop straw.Saw dust, husk, agricultural crop straw play the absorption microbial inoculum, keep moisture, loose ventilative, the effect such as C, N source is provided for functional microorganism.Preferably saw dust, rice husk, stalk etc. were tanned by the sun under the sun 1～2 day before making fermentation bed, column home is wanted thorough disinfection.
Fermenting bed padding should without go mouldy, without corrupt, ordorless or other peculiar smell.Material particular diameter is at 2～10mm, carbon nitrogen ratio 40～100: 1, and colibacillus of excrement number≤100cfu/g, induced worm egg death rate 〉=98%, pH value 7.5 left and right.Chemical glue plywood saw dust can not be used as bedding and padding.
In fermentation vat, product of the present invention, bedding and padding, assisted fermentation agent being added water mixes, be layered in fermentation vat, moisture content 45% left and right of bedding and padding, about heap fermentation 3 days, plough deeply once when bedding and padding middle levels temperature reaches 45 ℃～55 ℃, then ferment about 3 days, when the middle level temperature is down to 40 ℃ of left and right, the bedding and padding of piling up are raked, meet the requirements and just can advance pig, undesirable fermentation sterilization again.After the fermenting bed padding compacting, thickness is 80cm, than the low 10cm of the table top of searching for food left and right.
2. test grouping
Carried out product fermentation bed cultivation technology pig-keeping experiment of the present invention 5 days～December 8 July in 2009 on certain pig farm.Bedding and padding are made of by the weight ratio of 1: 1 saw dust, rice husk.By term, the principle that body weight is close, select same kind, with batch, 180 of hybridization (the local white ♀ * Yorkshire ♂) pigs of mean body weight (16.35 ± 2.33) kg, every pig hangs up ear tag, is divided at random 3 groups, every group of 4 repetition, 8 boars of every repetition, 7 sows.Test I group is control group, traditional concrete floor hutch; Test II group, the fermentation bed cultivation made from complex micro organism fungicide of the present invention, microbial inoculum component and weight percent thereof consist of: fermentation of bacillus subtilis medicinal powder 35%, the lichen bacillus ferments medicinal powder 35%, Candida utilis fermented product pulvis 30%, the consumption 200g/ cubic meter bedding and padding of starter; Test III group, the fermentation bed cultivation made from complex micro organism fungicide of the present invention, microbial inoculum component and weight percent thereof consist of: fermentation of bacillus subtilis medicinal powder 40%, the lichen bacillus ferments medicinal powder 40%, Candida utilis fermented product pulvis 20%, the consumption 200g/ cubic meter bedding and padding of starter.
3. experimental result is as follows:
Table 1 colony house environment and live pig surface observation result
Table 2 live pig production performance result
Can be found out by table 1, table 2, it is good that the colony house environment that test II, III organize is obviously organized than contrast I, without obvious stink, grows without obvious mosquitos and flies; The average daily gain of test II, III group is higher than contrast I group (P＜0.05), and feedstuff-meat ratio is lower than control group (P＜0.05), and test II, III group difference is not remarkable.Reach a conclusion thus, microbial inoculum of the present invention can alleviate environmental pollution, improves efficiency of feed utilization, effectively promotes the growth of pig.
The impact of table 3 raising pattern on the live pig meat
As can be seen from Table 3, the test group meat quality is better than control group.Simultaneously pork is checked through Safety of Livestock Products ' Quality supervision and inspection center of the Ministry of Agriculture (Changsha), each test group pork meat meets " no public nuisance livestock meat safety requirements standard (GB18406.3-2001) " requirement fully, illustrates that fermentation bed to raise pig has no adverse effects to the quality of pork.
Table 4 bedding and padding temperature variation
As can be seen from Table 4, under test II, III group bedding and padding top layer the 5cm place, and mean daily temperature is respectively 28.5 ℃, 29.0 ℃, and difference is not remarkable; The mean daily temperature at 30cm place, test II group is 40.9 ℃, and test III group is 44.3 ℃, and the bedding and padding nexine keeps higher temperature, is conducive to the rapid degraded of pig manure urine.
In table 5 bedding and padding, useful viable count changes
As can be seen from Table 5, when fermentation entered 30d, it was 10 that test II group is counted content with III group probiotics
8The order of magnitude; During fermentation 150d, test II group is counted content with III group probiotics and is still kept 10
7The order of magnitude illustrates that product of the present invention has stronger colonization ability in fermenting bed padding.
Claims (6)
1. the preparation method of a high-efficiency fermenting bed use complex micro organism fungicide, is characterized in that, comprises the following steps:
1) subtilis ACCC 10619 fermented product preparations
At first, subtilis is cultivated through slant strains activation, shake-flask seed;
Then carry out fermentor cultivation: in fermentor tank, the substratum loading amount is 50～70% of cumulative volume, inoculum size accounts for 0.1～0.3% of culture volume, 34～37 ° of C of leavening temperature, blowing air 180～200rpm stir culture, ratio 0.9:1～the 1:1 of ventilation volume and fermentating liquid volume, tank pressure 0.03～0.05MPa; Fermentation termination be the gemma number be not less than total count 90%, and the viable count of fermented liquid is 1.0 * 10
10～1.5 * 10
10Between cfu/mL; Each constituent mass per-cent of fermentation tank culture medium is: glucose 0.5～0.7%, starch 0.3～0.5%, soybean cake powder 0.8～1.2%, manganous sulfate 0.0062%, yeast powder 0.5～0.8%, peptone 0.3～0.5%, iron(ic) chloride 0.01%, potassium primary phosphate 0.4%, calcium carbonate 0.1%, sal epsom 0.05%, all the other are water, pH7.2～7.4;
Carry out at last solid absorption: fermented liquid is pressed the part by weight absorption of 1:2～1:3 with wheat bran, after absorption, low temperature dehumidifier drying, pulverize, and the powder product bacteria containing amount is 2.0 * 10
9～5.0 * 10
9Between cfu/g;
2) the lichen bacillus ferments thing preparation
At first, Bacillus licheniformis is cultivated through slant strains activation, shake-flask seed; Described Bacillus licheniformis, preserving number are CGMCC No.4835;
Then carry out fermentor cultivation: in fermentor tank, the substratum loading amount is 50～70% of cumulative volume, inoculum size accounts for 0.1～0.3% of culture volume, 45～50 ° of C of leavening temperature, blowing air 180～200rpm stir culture, ratio 0.9:1～the 1:1 of ventilation volume and fermentating liquid volume, tank pressure remains on 0.03～0.05MPa, fermentation termination be the gemma number be not less than total count 90%, and fermented liquid contain the bacterium number 8.0 * 10
9～1.2 * 10
10Between cfu/mL; Each constituent mass per-cent of fermentation tank culture medium is: starch 1.0～1.5%, dextrin 0.5～0.8%, soybean cake powder 1.2～1.5%, ammonium sulfate 0.6～0.8%, yeast powder 0.3～0.4%, calcium carbonate 0.2%, sodium-chlor 0.4%, potassium primary phosphate 0.02%, all the other are water, pH7.0～7.2;
Carry out at last solid absorption: fermented liquid is pressed the part by weight absorption of 1:2～1:3 with wheat bran, after absorption, low temperature dehumidifier drying, pulverize, and the powder product bacteria containing amount is 1.5 * 10
9～4.0 * 10
9Between cfu/g;
3) Candida utilis ACCC 20060 fermented product preparations
At first, Candida utilis is cultivated through slant strains activation, shake-flask seed;
Then carry out fermentor cultivation: in fermentor tank, the substratum loading amount is 50～70% of cumulative volume, inoculum size accounts for 0.3～0.5% of culture volume, 25～28 ° of C of leavening temperature, blowing air 120～150rpm stir culture, ratio 1:1～the 1.1:1 of ventilation volume and fermentating liquid volume, tank pressure remains on 0.03～0.05MPa, 46～48 hours complete secondary fermentation liquid that ferments contain the bacterium number 2.0 * 10
9～3.0 * 10
9Between cfu/mL; Each constituent mass per-cent of fermentation tank culture medium is: glucose 3.5～4.0%, and ammonium sulfate 1.5～2.0%, yeast powder 0.2～0.25%, potassium primary phosphate 0.1%, sodium-chlor 0.01%, sal epsom 0.05%, all the other are water, pH6.0～6.5;
Carry out at last solid absorption: fermented liquid is pressed the part by weight absorption of 1:1 with wheat bran, after absorption, low temperature dehumidifier drying, pulverize, and the powder product bacteria containing amount is 1.5 * 10
9～2.5 * 10
9Between cfu/g;
4) high-efficiency fermenting bed is composite with complex micro organism fungicide
Careless fermentation of bacillus medicinal powder 30～40% by weight percentage, the lichen bacillus ferments medicinal powder 30～40%, Candida utilis fermented product pulvis 20%～30%, fully mixing gets final product;
Step 2) each constituent mass per-cent of substratum of described slant strains activation is: glucose 0.4%, and malt extract 1%, yeast extract 0.4%, calcium carbonate 0.2%, agar 1.5%～2.0%, all the other are water, pH7.0～7.2;
Step 2) substratum cultivated of described shake-flask seed is in step 2) do not add agar in described slant medium, concrete operations: fresh slant strains one ring of getting activation, be inoculated in shake-flask seed liquid substratum, loading amount is 100mL/500mL, rotating speed is 180～200rpm, after 45～50 ℃ of constant temperature culture 28～30h, stop cultivating.
2. method according to claim 1, is characterized in that, described high-efficiency fermenting bed is not less than 1.0 * 10 with the total viable count of complex micro organism fungicide
9Cfu/g.
3. method according to claim 1, is characterized in that,
Each constituent mass per-cent of substratum of the described slant strains activation of step 1) is: peptone 1%, and NaCl0.5%, yeast extract paste 0.5%, agar 1.5%～2.0%, all the other are water, pH7.2-7.4;
The substratum that the described shake-flask seed of step 1) is cultivated is not add agar in the described slant medium of step 1), concrete operations: fresh slant strains one ring of getting activation, be inoculated in shake-flask seed liquid substratum, loading amount is 100mL/500mL, rotating speed is 180～200rpm, after 34～37 ℃ of constant temperature culture 22～24h, 80 ° of C water-bath thermal treatment 10 minutes gets final product.
4. method according to claim 1, is characterized in that,
The substratum of the described slant strains activation of step 3) is the PDA substratum;
The described shake-flask seed of step 3) is cultivated concrete operations: fresh slant strains one ring of getting activation, be inoculated in shake-flask seed liquid substratum, loading amount is 150mL/500mL, and rotating speed is 120～150rpm, after 25～28 ℃ of constant temperature culture 30～32h, stop cultivating; Each constituent mass per-cent of the substratum of described shake-flask seed: glucose 2%, peptone 2%, yeast extract 1%, all the other are water, pH6.0～6.5.
5. a microbial inoculum for high-efficiency fermenting bed, is characterized in that, is the microbial inoculum that is prepared from by the described method of claim 1-4 any one.
6. a strain is used bacterial strain for microbial inoculum for high-efficiency fermenting bed fermentation preparation, it is characterized in that, described bacterial strain is Bacillus licheniformis (Bacillus licheniformis), and preserving number is CGMCC No.4835.
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