CN100408673C - Organic microbial composite and use - Google Patents

Organic microbial composite and use Download PDF

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CN100408673C
CN100408673C CNB2006100374306A CN200610037430A CN100408673C CN 100408673 C CN100408673 C CN 100408673C CN B2006100374306 A CNB2006100374306 A CN B2006100374306A CN 200610037430 A CN200610037430 A CN 200610037430A CN 100408673 C CN100408673 C CN 100408673C
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solid medium
fermentation
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CN1955276A (en
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利耀铭
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Abstract

This invention provides an organic microorganism composite and its usage, and it relates to biotechnology field. The strain of this composite includes trichoderma aspergillus, actinomycetes, bacillus and saccharomyces cerevisiae Li1012. The composite whose bacterial number, yeast count and mycetes number are 1.0-4.0*108cfu/g, 3.0-8.0*105cfu/g and 2.0-5.5*106cfu/g is prepared after the process of pure mass-cultivation, liquid fermentation and solid fermentation, with its water content lower than 10%, which can be used in producing organic fertilizer or forage additives. It will be of great contribution towards the health of crops when it is used for fertilizer preparation.

Description

Organic microbial composite and purposes
Technical field
The present invention relates to biological technical field, be specifically related to a kind of organic microbial composite and purposes that contains the living microorganism flora.
Background technology
The factory that pushes away that more furthers investigate with suitability for industrialized production along with microorganism application technology uses, the application of microorganism technology has had develops by leaps and bounds, particularly after recognizing to use chemical fertilizer and agricultural chemicals in a large number, people cause the pollution of environment and the influence of agricultural product quality, long-term excessive use chemical fertilizer, agricultural chemicals, caused soil compaction, ecotope is unbalance, harmful chemical residue severe overweight, water pollutes, cause the quality of agricultural-food to descend, the food flavor mouthfeel is poor, multiple due trace element and nutrition leak.Equally, the poultry and livestock feed of producing with these farm crop can not make growth healthily after the feed of fowl poultry again, so taked means such as hormone, microbiotic again, formed vicious cycle, finally make the agricultural-food of China be in low level, low-level, low price for a long time, outlet more is subjected to more and more higher barrier restriction.Aspect edible, because exceeding standard of the due trace element of famine and nutrition and objectionable impurities seriously damaged the immune and healthy of consumers in general.Therefore, the application of microorganism technology deeply is applied to agricultural.
The composition that contains the living microorganism flora is a kind of beneficial microorganism colony that can produce multiple catalytic decomposition enzyme, and it contains useful bacterium (Bacteria), yeast (Yeast), mould (Mold) three major types microorganism, and powerful fermentation capacity of decomposition is arranged.Detecting by food hygiene law is mushroom to the person poultry harmless, and it has and can secrete a large amount of powerful saccharifying enzyme, lipolytic enzyme, cellulolytic enzyme, sucrose decomposition enzyme, alcohol lytic enzyme, lactose decomposing enzyme, oxydo-reductase, maltose lytic enzyme, urea decomposing enzyme etc., very strong aerobic fermentation ability is arranged, just can bring into play decomposition in short duration, the effect of catalysis organic matter is decomposed and is eliminated various objectionable impuritiess.
The various fermentation fertilizers that contain the composition of living microorganism flora, be used for plant husbandry, can not only provide sufficient isostatic nutrition to farm crop, and can also rely on the powerful fermentation capacity of decomposition of ferment flora, and progressively eliminate the pollution of chemical fertilizer, agricultural chemicals for soil, product, transform soil, reach green, organic standard, the composition that contains the living microorganism flora wherein can continue to play a role in the soil midium or long term, thereby plays increasing both production and income, improves the effect of quality.
The composition fodder additives that contains the living microorganism flora is used for breed domestic animal is played well whole intestines effect, harmful bacterium activity is in vivo had inhibition and stays dead effect, thereby play very big effect to keeping fit.Owing to contain the effect of the composition and the enzyme thereof of living microorganism flora, feed is digestion well easily in the digestion organs of domestic animal, has improved the digestibility of food, reduces the stink of ight soil, reduces animal house, and the hen house stink is to the public hazards of environment.
Relevant report contains the fertilizer of living microorganism, the patent and the document of fodder additives are a lot, they all are to select some different profitable strains separately, carry out fermentation culture and prepare fertilizer, fodder additives, each fertilizer, the manufacturer of fodder additives all will set up the cultivation of a cover microorganism species, preserve, the equipment of fermentation is like this because the equipment of manufacturer, condition, the restriction of ability etc., the fertilizer of being produced, fodder additives does not satisfy the needs of wide agriculture production, so we are necessary to develop the composition that contains the living microorganism flora that is fit to China's actual conditions, use said composition as long as use the mode of compost, do not need the cultivation of special microorganism species, preserve, the equipment of fermentation, do not need factory just can prepare organic fertilizer and fodder additives yet, help applying in vast rural area.
Summary of the invention
The object of the present invention is to provide a kind of organic microbial composite that contains the living microorganism flora of suitable China's actual conditions, can be used for preparing fertilizer, fodder additives.
For achieving the above object, a kind of organic microbial composite provided by the present invention, be that to adopt Trichoderma (Trichoderma), aspergillus tubigensis (Aspergillus niger), actinomycetes (Actinomyces), genus bacillus (Bacillus subtilis Bacillus licheniformis), S. cervisiae Li1012 (Saccharomyces ceresisias Li1012) be bacterial classification, contain bacterial count 1.0~4.0 * 10 with what be prepared as follows that step makes 3Cfu/g, yeast count 3.0~6.0 * 10 5Cfu/g, fungi count 2.0~5.5 * 10 6The composition of cfu/g, its water content≤10%, described preparation process:
The purebred enlarged culturing of the first step: A placed transparent vessel with solid medium, and sterilization is got Trichoderma, aspergillus tubigensis, actinomycetes, genus bacillus bacterial classification respectively and is inoculated in respectively among the solid medium A, in 30~33 ℃ of constant temperature culture 48~72 hours;
The second step liquid fermentation and culture: get S. cervisiae Li1012 CCTCC No.M206044 bacterial classification inoculation in liquid nutrient medium, cultivated 18~20 hours in 30~32 ℃ of constant-temperature shaking culture or under 150 rev/mins rotating speed under the logical oxygen fermented liquid;
The 3rd step solid fermentation is cultivated: add solid medium B in fermentor tank, get solid medium B weight 0.06~0.2% through the fermented liquid of the second step liquid fermentation and culture gained and respectively get among the solid medium B of 0.1~0.3% Trichoderma, aspergillus tubigensis, actinomycetes, genus bacillus access fermentor tank of solid medium B weight through the purebred enlarged culturing of the first step, in 31~34 ℃ of temperature, cultivate and got solid culture in 30~36 hours;
The 4th step dry below 70 ℃, was controlled water content≤10% of resulting composition with the 3rd solid culture that goes on foot gained.
The solid medium A that adopts during the purebred enlarged culturing of the above-mentioned the first step can adopt the raw material of following weight part to form: wheat bran 80, Semen Maydis powder 20, water 70~80 or wheat bran 80, Semen Maydis powder 10, ground rice 10, water 70~80 etc., and the preferred raw material of 100 parts in wheat bran, water 70~80 parts (weight parts) that adopts is formed in order to save production cost;
The second step liquid fermentation and culture: said liquid nutrient medium can adopt following raw materials by weight percent to form: yeast extract paste 0.5~0.8%, peptone 1.0~1.3%, glucose 0.5~0.8%, white sugar 1.0~1.3%, potassium primary phosphate (KH 2PO 4) 0.12~0.15%, all the other are water; Preferred employing pH value is 6.4~7.0, is made up of following raw materials by weight percent:
Yeast extract paste 0.5~0.8% peptone 1.0~1.3% glucose 0.5~0.8%
All the other are water for white sugar 1.0~1.3%.
When the 3rd step, solid fermentation was cultivated: said solids substratum B, can form with following raw materials by weight percent:
1,45 parts in wheat bran, 40 parts in rice bran, 5 parts of soybean cake powders, 10 parts of Semen Maydis powder, 45~50 parts in water;
2,50 parts in wheat bran, 40 parts in rice bran, 10 parts of Semen Maydis powder, 45~50 parts in water; The preferred raw material of following weight part that adopts is formed in order to save production cost:
45~55 parts in 10~5 parts of water of 90~95 parts of rice brans of wheat bran
For the output that improves composition and convenient the use, said the 3rd step solid fermentation is cultivated and can be adopted second order fermentation, promptly further carries out second order fermentation cultivate the resulting composition that contains the living microorganism flora through the one-level solid fermentation after.The composition of getting the living microorganism flora of one-level solid fermentation cultivation gained inserts among the solid medium C by 1% of solid medium C weight and carries out fermentation culture, 45~60 ℃ of fermentation culture temperature, look material leavening temperature situation and carry out the turning ventilation, generally once beginning turning in postvaccinal 24 hours, later on once every turning in 12 hours, cultivate after 72~96 hours dry below 70 ℃, the water content of control combination thing≤10%, gained contain and contains bacterial count in the composition of living microorganism flora is 1.0~2.0 * 10 8Cfu/g, yeast count are 3.0~4.0 * 10 5Cfu/g, fungi count are 2.0~3.0 * 10 6Cfu/g;
Form for the preferred raw material of following weight part that adopts that saves production cost, said solid medium C can adopt following raw materials in part by weight to form:
50 parts in wheat bran, 48 parts in rice bran, 2 parts of Semen Maydis powder, 1.5 parts in brown sugar, 50~55 parts of moisture content; Preferred 50 parts in wheat bran, 48 parts in rice bran, 2 parts in brown sugar, the moisture content of adopting is made solid medium C for 50~55 parts.
The used S. cervisiae Saccharomyces cerevisiae bacterium of the present invention number is Li1012, be preserved in Chinese Wuhan University China typical culture collection center on April 30th, 2006, preserving number CCTCCNO:M 206044, the thalline of this bacterial strain during than the strain growth of saccharomyces cerevisiae is abundant, moistening, preservation period is stable, enter when being used to produce produce the alcohol state than the bacterial strain of saccharomyces cerevisiae fast again more than.
Trichoderma (Trichoderma), aspergillus tubigensis (Aspergillus niger), actinomycetes (Actinomyces), genus bacillus (Bacillus subtilis Bacillus licheniformis) are known probioticss.
Genus bacillus is a kind of aerobic non-pathogenic bacteria, it exists with interior form of giving birth to son at enteron aisle or storage middle part, do not consume feed nutrient, can keep quality of the fodder, after entering enteron aisle, bring back to life rapidly at enteron aisle, the resurrection rate is near 100%, it can make the pH value in the jejunum descend, and ammonia concentration reduces, and consumes a large amount of oxygen, keep the enteron aisle anaerobic environment, suppress pathogen growth, keep the normal eubiosis of enteron aisle, it can produce active very high proteolytic enzyme, amylase, lipase and multiple amino acids.It is strong that genus bacillus anti-presses down property, has characteristics such as high temperature resistant, acid and alkali-resistance.
Aspergillus tubigensis can produce the very strong enzyme system of various active as glucoamylase, αDian Fenmei, proteolytic enzyme, cellulase etc. in metabolic process.Can improve the digestibility of feed and the transformation efficiency of nutrition.
Trichoderma is the important bacterial classification of production of cellulose enzyme, and complex materials such as decomposition of cellulose and xylogen quicken cellulosic decomposition such as tangerine bar, rice husk, wheat bran in the feed strongly, improve the digestibility and the utilization ratio of feed.
Actinomycetes are main generation bacterium of natural antibiotics, can improve the resistance against diseases of fowl poultry.
Organic microbial composite provided by the present invention can be used for preparing organic fertilizer or fodder additives.
The present invention has the following advantages:
1. the present invention has used novel S. cervisiae, and the composition of the living microorganism flora of gained can produce senior alcohol quickly and efficiently in the process that is used for preparing fertilizer, fodder additives.Provide good nutrition source for producing the actinomycetes, the lactobacillus that suppress pathogenic bacteria, thereby breeding efficiently, being used to prepare fertilizer has great effect to the healthy growths of farm crop, and the output of farm crop is improved greatly, the quality of product increases, and mouthfeel improves.Be used to prepare fodder additives the healthy growth of fowl poultry is also had great effect
2. product preparation method of the present invention is simple, and operational condition is easy to control, the culture medium raw material wide material sources, and cheap, with short production cycle, production cost is low, favourable applying.
3. quality product height.The present invention adopts the microbial liquid fermentation technique that combines with solid, can better solve in the solid solid fermentation process, and fermentation period is long, pollutes easily, and various fermentation conditions are difficult to control problem.Guaranteed quality product.
Further set forth technical scheme of the present invention and beneficial effect below by specific embodiment
Embodiment
Source, cultivation and the preservation of embodiment 1 S. cervisiae Li1012 (Saccharomyces cerevisiae Li1012)
The source: isolation and selection from the soil in the lichee orchard of Guangdong Baiyun District plantation and the wild yeast bacterium.
Being accredited as S. cervisiae Li1012 through Chinese Wuhan University China typical culture collection center cultivates:
The substratum that adopts: with peeling potatoes, be cut into small pieces, added water boil 30~45 minutes, filter, the potato after filtering is blended under the normal atmosphere according to weight ratio with glucose and water at 10: 1: 50 sterilized 18~30 minutes.
Strain culturing: S. cervisiae Li1012 is inoculated in the above-mentioned substratum, place 32~35 ℃ incubator cultivated 3~5 days barms.
Bacterial strain is preserved:
The slant culture based formulas: all the other are water for yeast extract paste 0.5~0.8% peptone 1.0~1.3% glucose 0.5~0.8% white sugar 1.0~1.3% agar 2% PH=6.4.A normal atmosphere was sterilized 20 minutes.
Substratum is made slant tube, after streak inoculation is cultivated, place 3~5 ℃ of refrigerators and preserve.
The preparation of embodiment 2 organic microbial composites
Step 1: (1) purebred enlarged culturing: make solid medium A after taking by weighing wheat bran 1000g water 700g stirring and evenly mixing, be divided in the triangular flask of 500ml, the culture medium A of every bottled 300g, the bacterial classification of getting Trichoderma, aspergillus tubigensis, actinomycetes, genus bacillus 0.1MPa sterilize in 30 minutes respectively is inoculated in the triangular flask that substratum is housed under aseptic condition respectively, place thermostat container to cultivate, cultivated 72 hours at 30~33 ℃;
(2) liquid fermentation and culture:
Take by weighing after yeast extract paste 5g, peptone 10g, grape sugar 8g, white sugar 12g, the water 965g stirring and dissolving pH value and be 6.5 and make liquid nutrient medium, be divided in the triangular flask of 500ml, every bottled 250ml, 0.1MPa sterilize 30 minutes, the yeast saccharomyces cerevisiae bacterial classification of getting embodiment 1 is inoculated in the triangular flask that liquid nutrient medium is housed under aseptic condition, place shaking table, got fermented liquid in 20 hours in 30~32 ℃ of constant-temperature shaking culture.
(3) use 5M 3Fermentor tank carries out solid fermentation and cultivates:
Taking by weighing 450 kilograms in wheat bran, 50 kilograms in rice bran, the water yield is 250kg, put to place behind the stirring and evenly mixing in the fermentor tank and made solid medium in 2 hours, 0.1MPa sterilize after 30 minutes, put and be chilled to 40 degree, the fermented liquid of the S. cervisiae of 300g Trichoderma, 300g aspergillus tubigensis, 300g actinomycetes, 300g genus bacillus and the 1000g of the above-mentioned cultivation of adding under aseptic condition, stir, regulate the fermentor tank air flow to 0.5 cubic meters per minute, controlled temperature was cultivated 36 hours at 31~34 ℃, and discharging gets solid culture;
(4) solid culture of gained is dry below 70 ℃, and the water content of resulting composition is 5.8%, and the bacterial count that gets said composition by GB 4789-94 microbiological test of food hygiene standard and NY/T 227-94 standard test is 3.0 * 10 8Cfu/g, yeast count are 7.0 * 10 5Cfu/g, fungi count are 5.0 * 10 6Cfu/g
The preparation of embodiment 3 organic microbial composites
(1) purebred enlarged culturing: make solid medium A after taking by weighing wheat bran 2000g water 1500g stirring and evenly mixing, be divided in the triangular flask of 500ml, the culture medium A of every bottled 250g, the bacterial classification of getting Trichoderma, aspergillus tubigensis, actinomycetes, genus bacillus 0.1MPa sterilize in 30 minutes respectively is inoculated in the triangular flask that substratum is housed under aseptic condition respectively, place thermostat container to cultivate, cultivated 48 hours at 30~33 ℃;
(2) liquid fermentation and culture:
Take by weighing after yeast extract paste 80g, peptone 130g, grape sugar 50g, white sugar 100g, the water 9640g stirring and dissolving pH value and be 6.5 and make liquid nutrient medium, be loaded in the fermentor tank of 5L, 0.1MPa sterilize 30 minutes, getting the yeast saccharomyces cerevisiae bacterial classification of embodiment 1 inoculates under aseptic condition, under 150 rev/mins rotating speed under the logical oxygen stirs, control 30~32 ℃, cultivate 18 hours must fermented liquid.
(3) use 10M 3Fermentor tank carries out solid fermentation and cultivates:
Taking by weighing 950 kilograms in wheat bran, 50 kilograms in rice bran, the water yield is 500kg, put to place behind the stirring and evenly mixing in the fermentor tank and made solid medium in 2 hours, 0.1MPa sterilize after 30 minutes, put and be chilled to 40 degree, the fermented liquid of the S. cervisiae of 1500g Trichoderma, 1500g aspergillus tubigensis, 1500g actinomycetes, 1500g genus bacillus and the 3000g of the above-mentioned cultivation of adding under aseptic condition, stir, regulate the fermentor tank air flow to 0.5 cubic meters per minute, controlled temperature was cultivated 36 hours at 31~34 ℃, and discharging gets solid culture;
(4) solid culture of gained is dry below 70 ℃, and the water content of resulting composition is 8.8%, and the bacterial count that gets said composition by GB/T 4789-94 and NY/T 227-94 standard test is 4.0 * 10 8Cfu/g, yeast count are 8.0 * 10 5Cfu/g, fungi count are 5.5 * 10 6Cfu/g
(5) secondary solid fermentation:
Take by weighing 480 kilograms in rice bran, 500 kilograms in wheat bran, 20 kilograms in brown sugar, water and make solid medium C for 550 kilograms, other takes by weighing 10 kilograms of the compositions of above-mentioned (4) gained
The preparation method:
1. brown sugar is dissolved in 20 kilograms more than 90 ℃ in the hot water, makes its whole dissolvings, pour in the clean clear water of outfit standby again through stirring.
2. rice bran, wheat bran and original seed are poured into abundant mixing in the container.
3. brown sugar water is evenly added abundant mixing in the raw material.
4. the mixture that will mix and stir is poured in the fermenter, and bed thickness is the 40-50 centimetre, and the surface is wipeed off, covers gunnysack and carries out spontaneous fermentation, and controlled temperature is no more than 60 ℃.
5. turning in per during the fermentation 24 hours once, ferment altogether 120 hours immediately with material dry below 70 ℃.The composition measuring water content of gained is 5.8%, and the bacterial count that gets said composition by GB/T 4789-94 and NY/T 227-94 standard test is 1.8 * 10 8Cfu/g, yeast count are 4.0 * 10 5Cfu/g, fungi count are 2.8 * 10 6Cfu/g
The preparation of embodiment 4 fermentation fertilizers
A. get 2 kilograms in brown sugar and add 20 kg of water to boil the back cool to below 40 ℃, pour in 10 kilograms of the microbial composites of 48 kilograms in rice bran and embodiment 3, and mix;
B, raw material and mountain soil 450 kilograms, 450 kilograms of mixing of water that 100 kilograms of bone meal, 300 kilograms of chicken manures, 100 kilograms of dregs of beans, 50 kilograms of fish meal etc. are needed;
C, pile chevron after a and b are mixed again and carry out spontaneous fermentation;
D. once, every turning in 24 hours fermented altogether five days after 70 following dryings can make fertilizer and use.
Contain bacterial count 5.0 * 10 in the prepared fertilizer 7Cfu/g, yeast count 4.0 * 10 4Cfu/g, fungi count 2.0 * 10 4Cfu/g
The effect of fertilizer test that embodiment 5 Application Examples 4 are provided
The fermentation fertilizer that adopts embodiment 4 to make, it is as follows to be used to plant balsam pear and other fertilizer simultaneous test:
The experiment date: beginning on September 1st, 2003
Detect the date: on December 5th, 2003
Test method: be used for the experiment that balsam pear is planted with 4 kinds of different fertilizer, 4 kinds of fertilizer carry out six parallel processing altogether, and three repetitions are carried out in each processing, carry out the mensuration of test item after each sample plot results.The method of fertilising is identical with quantity.
Test sample:
Sample 1: use embodiment 4 fermentation fertilizers of the present invention
Sample 2: use the Guangzhou research of agricultural science and spend eastern planting experiment base self-control farm organic fertilizer
Sample 3: use commercially available hundred health compoiste fertilizers
Sample 4: use compound organic and inorganic fertilizer
Test-results:
Figure C20061003743000111
Conclusion
From detected result as can be seen, in the balsam pear planting experiment that carries out with 4 kinds of different fertilizer, the balsam pear of planting out is remaining basically stable aspect reducing sugar, crude protein, the Vitamin C content, and difference is not obvious, but other fertilizer of output average specific of sample 1 exceeds 13.8%.
The preparation of embodiment 6 fodder additivess
1. material: 8 kilograms of the microbial composites of 3 kilograms in brown sugar, embodiment 3,40 kilograms of bone meal, 20 kilograms of dregs of beans, 30 kilograms of fish meal, 1000 kilograms in clean mountain soil;
2. brown sugar adds 450 kilograms in water to boil the back cool to below 40 ℃, is poured on the rice bran that mixes, the microbial composite, and stirs and 40 kilograms of bone meal, 20 kilograms of dregs of beans, 30 kilograms of fish meal, 1000 kilograms of mixings of mountain soil again;
3. the material of top mixing is placed fermenter to pile up, the thickness of the bed of material be after 40~50 centimeters every turning in 24 hours once, ferment altogether five days can be 70 ℃ of dryings
Contain bacterial count 5.0 * 10 in the fodder additives that this fermentation is finished 7Cfu/g, yeast count 4.0 * 10 4Cfu/g, fungi count 2.0 * 10 4Cfu/g
Embodiment 7 usefulness fodder additivess are fed piggy step-by-step test situation
Test method: 30 of piggys, be divided into three groups at random, 10 every group, wherein 7 of boars, 3 of sows.Nursing time, mode, condition are identical
Test group 1: any mixture such as the embodiment of the invention 6 feed additives 2%, other 98% Semen Maydis powder, wheat bran, rice bran, dregs of beans are made forage feed
Test group 2: any mixture such as the embodiment of the invention 6 feed additives 4%, other 96% Semen Maydis powder, wheat bran, rice bran, dregs of beans are made forage feed
Control group 3: make forage feed with any mixture such as Semen Maydis powder, wheat bran, rice bran, dregs of beans
Test-results:
TBW is tested average every day weight gain diarrhoea of back TBW weightening finish before feeding the test of fate piggy
It Kg Kg Kg g head
Test group 1 26 220.5 418.25 197.75 760.6 0
Test group 2 26 217.0 421.00 204.00 784.6 0
Control group 26 220.5 398.50 178.00 685.0 4
Conclusion: feed piggy as pig feed addictive, the red dysentery characterized by white mucous stool of preventing and treating piglet effectively reduces diarrhea rate 40%) more than, improving piglet grain transformation efficiency 17%, day weight gain improves 20%, improves more than 14% than control group.

Claims (5)

1. organic microbial composite, it is characterized in that: be that to adopt Trichoderma, aspergillus tubigensis, actinomycetes, genus bacillus, S. cervisiae Li1012 (Saccharomyces cerevisiae Li1012) be bacterial classification, contain bacterial count 1.0~4.0 * 10 with what be prepared as follows that step makes 8Cfu/g, yeast count 3.0~8.0 * 10 5Cfu/g, fungi count 2.0~5.5 * 10 6The composition of cfu/g, its water content≤10%, described preparation process:
The purebred enlarged culturing of the first step: A places transparent vessel with solid medium, sterilization, the bacterial classification of getting Trichoderma, aspergillus tubigensis, actinomycetes, genus bacillus respectively is inoculated in respectively among the solid medium A, in 30~33 ℃ of constant temperature culture 48~72 hours, said solid medium A was made up of the raw material of following weight part: 80 parts in (1) wheat bran, 20 parts of Semen Maydis powder, 70~80 parts in water; Or 80 parts in (2) wheat bran, 10 parts of Semen Maydis powder, 10 parts in ground rice, 70~80 parts in water; Or 100 parts in (3) wheat bran, 70~80 parts in water;
The second step liquid fermentation and culture: the bacterial classification inoculation of getting S. cervisiae Li1012CCTCC No.M206044 is in liquid nutrient medium, in 30~32 ℃ of constant-temperature shaking culture or under logical oxygen, 150 rev/mins rotating speed is cultivated down and was got fermented liquid in 18~20 hours, and said liquid nutrient medium is made up of following raw materials by weight percent: (1) yeast extract paste 0.5~0.8%, peptone 1.0~1.3%, glucose 0.5~0.8%, white sugar 1.0~1.3%, potassium primary phosphate (KH 2PO 4) 0.12~0.15%, all the other are water; Or (2) yeast extract paste 0.5~0.8%, peptone 1.0~1.3%, glucose 0.5~0.8%, white sugar 1.0~1.3% all the other be water, the pH value of substratum is 6.4~7.0;
The 3rd step solid fermentation is cultivated: add solid medium B in fermentor tank, get solid medium B weight 0.06~0.2% through the fermented liquid of the second step liquid fermentation and culture gained and respectively get among the solid medium B of 0.1~0.3% Trichoderma, aspergillus tubigensis, actinomycetes, genus bacillus access fermentor tank of solid medium B weight through the purebred enlarged culturing of the first step, in 31~34 ℃ of temperature, cultivate and got solid culture in 30~36 hours
Said solid medium B is made up of the raw material of following weight part: 45 parts in (1) wheat bran, 40 parts in rice bran, 5 parts of soybean cake powders, 10 parts of Semen Maydis powder, 45~50 parts in water; Or 50 parts in (2) wheat bran, 40 parts in rice bran, 10 parts of Semen Maydis powder, 45~50 parts in water; Or 90~95 parts in (3) wheat bran, 5~10 parts in rice bran, 45~55 parts in water;
The 4th step dry below 70 ℃, was controlled water content≤10% of gained organic microbial composite with the 3rd solid culture that goes on foot gained.
2. organic microbial composite according to claim 1 is characterized in that:
The solid medium A that adopts during the purebred enlarged culturing of the first step is made up of the raw material of following weight part:
100 parts in wheat bran, 70~80 parts in water;
The second step liquid fermentation and culture: said liquid nutrient medium pH value is 6.4~7.0, is made up of following raw materials by weight percent:
Yeast extract paste 0.5~0.8% peptone 1.0~1.3% glucose 0.5~0.8%
All the other are water for white sugar 1.0~1.3%;
The 3rd step solid fermentation is cultivated: said solid medium B, be made up of the raw material of following weight part:
45~55 parts in 5~10 parts of water of 90~95 parts of rice brans of wheat bran.
3. organic microbial composite according to claim 1, it is characterized in that: said the 3rd step solid fermentation is cultivated and is adopted second order fermentation, after cultivating resulting organic microbial composite, further carry out second order fermentation through the one-level solid fermentation, the organic microbial composite of getting one-level solid fermentation cultivation gained inserts among the solid medium C by 1% of solid medium C weight to be cultivated, 45~60 ℃ of temperature, cultivate after 72~96 hours, dry below 70 ℃, it is 1.0~2.0 * 10 that the water content of control combination thing≤10%, gained organic microbial composite contain bacterial count 8Cfu/g, yeast count are 3.0~4.0 * 10 5Cfu/g, fungi count are 2.0~3.0 * 10 6Cfu/g, said solid medium C is made up of following raw materials in part by weight: 50 parts in (1) wheat bran, 48 parts in rice bran, 2 parts of Semen Maydis powder, 1.5 parts in brown sugar, 50~55 parts in water; Or 50 parts in (2) wheat bran, 48 parts in rice bran, 2 parts in brown sugar, water are made solid medium C for 50~55 parts.
4. organic microbial composite according to claim 3 is characterized in that: said solid medium C is made up of following raw materials in part by weight:
50~55 parts in 2 parts of water of 48 portions of brown sugar of 50 parts of rice brans of wheat bran.
5. the purposes of an organic microbial composite as claimed in claim 1 is to prepare organic fertilizer or fodder additives.
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