CN102199562B - Composite bacterial preparation as well as preparation method and application thereof - Google Patents
Composite bacterial preparation as well as preparation method and application thereof Download PDFInfo
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- CN102199562B CN102199562B CN2011100686474A CN201110068647A CN102199562B CN 102199562 B CN102199562 B CN 102199562B CN 2011100686474 A CN2011100686474 A CN 2011100686474A CN 201110068647 A CN201110068647 A CN 201110068647A CN 102199562 B CN102199562 B CN 102199562B
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Abstract
The invention discloses a composite bacterial preparation as well as a preparation method and application thereof, which belong to the technical field of agricultural biochemistry. The composite bacterial preparation comprises the following components in parts by volume: 10-40 parts of brevibacillus brives fermentation bacterial liquid, 10-40 parts of pseudomonas fluorescens fermentation bacterial liquid and 15-45 parts of sugarcane thermophilic actinomyces, wherein the bacterial count in each mol of fermentation liquid is 0.4-0.7 billion. The strains of brevibacillus brives, pseudomonas fluorescens and sugarcane thermophilic actinomyce are separated from rabbit dung and fungi residues; and fermentation bacterial liquids of the separated strains are mixed according to a proper proportion to form the composite bacterial preparation. The composite bacterial preparation disclosed by the invention can be used for accelerating the degradation of organic materials and reducing the conversion of organic nitrogen into gaseous nitrogen, so that the composite bacterial preparation can present stronger ammonia-suppressing and decomposing capabilities in the fermentation process of pig manure compost; and a smaller amount of ammonia gas is released while thorough decomposition of pig manure is accelerated.
Description
Technical field
The present invention relates to a kind of composite fungus agent and its preparation method and application, particularly a kind of composite fungus agent for the treatment of pig manure and its preparation method and application belongs to agricultural biological technical field.
Background technology
Along with the continuous large-scale development of aquaculture, a large amount of fowl and animal excrement serious environment pollutions of generation have also restricted expanding economy, have become the major issue of very important ecosystem environment protection.China's feces of livestock and poultry output had reached 36.4 hundred million t in 2000, was the same period 3.8 times of the industrial solid castoff amount, and the particularly a large amount of generation of swine excrement of fowl and animal excrement has caused showing great attention to of people.According to statistics, the day of 1 pig is drained fecaluria by 6kg, then is discharge feces 5 times of urine amount of people, produces fecaluria per year and reaches approximately 2.5t.Along with the day by day raising of living standards of the people, the livestock and poultry breeding industry of China will further develop in following 20 years, the pressure that will bring larger livestock and poultry to pollute.To the year two thousand twenty, the scale of the livestock and poultry breeding industry of China will be calendar year 2001 1.67 times of level, if the total amount of pollutant of when the time comes livestock and poultry breeding industry loss maintains level in 2000, loss coefficient then will be reduced to 60% of present level, and the environmental management pressure of livestock and poultry breeding industry is very huge.
On the other hand, remove in the feces of livestock and poultry and contain N, P, outside the K three elements, Cu is still arranged, Mg, S, Fe, the trace elements such as Zn, also be rich in some particular matter of stimulating plant growth, such as humin and microbiotic etc., it is the high-quality organic fertilizer resource that is rich in organic matter and nutrition nutrient, be organic fertilizer and the organic-inorganic compound mixed fertilizer that raw material is made with feces of livestock and poultry, it is a kind of complete manure, not only can improve the output of farm crop in the agricultural application, can also promote the formation of soil aggregate, make soil soft, increase the ventilation property of soil, water-permeable, thus the physicochemical character of soil improved.
The traditional method of utilizing feces of livestock and poultry to make organic fertilizer is compost, and feces of livestock and poultry is mixed with earth, mineral substance etc., under high temperature, many wet conditions, makes organic fertilizer through fermentation maturity, microorganism decomposition.In composting process, there is the volatilization of nitrogen, will reduce nitrogen content in the animal dung, reduce its fertilizer efficiency, thereby in the superior and inferior evaluating of compost quality, the preservation of nitrogen is a very important index.The nitrogen volatilization of feces of livestock and poultry mainly is the low molecule nitrogenous compound volatilization of fresh animal dung in storage process, mainly comprises ammonia (NH
3), Trimethylamine 99 [(CH
3)
3N], dimethylamine [(CH
3)
2NH] and Monomethylamine (CH
3N), ammonia wherein, Trimethylamine 99 are the important component parts of foul gas, and its volatilization also can serious environment pollution.In intensification and the pliotherm period of traditional pig manure, the pH value is higher, and deamination accounts for leading, the volatilization of nitrogen mainly is the release of ammonia, and simultaneously, the required decomposed period of traditional compost method is longer, generally need 1-2 month, be difficult to alleviate growing feces of livestock and poultry processing pressure.
The main path that addresses this problem both at home and abroad is that the using microbe fermenting agent is processed fowl and animal excrement production organic fertilizer, to shorten decomposed period.At present, the EM microbial inoculum (being combined by a certain percentage by useful mushrooms such as sporeformer, yeast, milk-acid bacterias) of Japan be generally acknowledged can quick composting a kind of composite fungus agent of feces of livestock and poultry.But this class microbial inoculum is when processing the fowl and animal excrement of high protein content class, and microorganism can produce a large amount of NH in the process that transforms nitrogen class material
4 +-N, NH
4 +-N, the Rapid Accumulation of these materials further improves the pH value, part ammonium nitrogen is further converted to nitric nitrogen and biomass attitude nitrogen by microorganism, another part then reaches more than 50 ℃ in heap temperature, pH in 7 the environment, volatilize with other most of volatile organic matters (such as thiomethyl alcohol, methylthio group etc.) in the mode of gaseous state.This is not only major cause and form that the compost stench produces, and is one of principal mode of compost nitrogen loss.According to research reports, the loss of this form accounts for 10%～100% of ammonium nitrogen.
Generally speaking, use and carry out high fibre content Composting with the EM microbial inoculum as the composite fungus agent of representative and can greatly shorten its decomposed period, but because pig manure is a kind of fowl and animal excrement of high protein content, quick, a large amount of decomposition of itrogenous organic substance matter can cause a large amount of ammonia volatilizations in digest process, a large amount of generations of ammonia are the severe contamination air not only, people and animals' health is brought great harm, and can greatly reduce compost quality owing to nitrogen loss.Therefore, this class microbial inoculum take the EM microbial inoculum as representative can not be applied to separately the Composting that pig manure etc. contains high protein.
Summary of the invention
The object of the invention is to overcome and utilize microbiobacterial agent to process existing above-mentioned deficiency in the feces of livestock and poultry process in the prior art, provide a kind of for new composite fungus agent and its preparation method and application.Composite fungus agent provided by the present invention can be used for processing the feces of livestock and poultry of the high protein content such as pig manure, and not only decomposed period is short, and deodorizing and nitrogen protecting is effective.
In order to realize the foregoing invention purpose, the invention provides following technical scheme:
A kind of composite fungus agent, it is comprised of the component of following volume ratio: (depositary institution is Chinese Typical Representative culture collection center to Brevibacillus brevis, the address is Wuhan, China Wuhan University, preservation date is on June 3rd, 2010, deposit number is CCTCC NO:M2010133, Classification And Nomenclature is Brevibacillus brevis WZX1 Brevibacillus brevis WZX1) 10～40 parts of zymocyte liquids, (depositary institution is Chinese Typical Representative culture collection center to Pseudomonas fluorescens, the address is Wuhan, China Wuhan University, preservation date is on June 3rd, 2010, deposit number is CCTCC NO:M2010135, and Classification And Nomenclature is Pseudomonas fluorescens WZX3
Pseudomonas fluorescensWZX3) (depositary institution is Chinese Typical Representative culture collection center for 10～40 parts of zymocyte liquids, thermoactinomyces sacchari, the address is Wuhan, China Wuhan University, preservation date is on June 3rd, 2010, and deposit number is CCTCC NO:M2010134, and Classification And Nomenclature is thermoactinomyces sacchari WZX2
Thermoactinomyces sacchariWZX2) zymocyte liquid is 15～45 parts, in the above-mentioned zymocyte liquid in every ml fermented liquid the bacterium number be 400,000,000-700,000,000.
The better of each zymocyte liquid volume ratio is chosen as in the above-mentioned composite fungus agent, 15～35 parts of Brevibacillus brevis zymocyte liquids, 15～35 parts of Pseudomonas fluorescens zymocyte liquids, 20～40 parts of thermoactinomyces saccharis.
The optimal selection of each zymocyte liquid volume ratio is in the above-mentioned composite fungus agent, 30 parts of Brevibacillus brevis zymocyte liquids, 30 parts of Pseudomonas fluorescens zymocyte liquids, 40 parts of thermoactinomyces sacchari zymocyte liquids.
In the above-mentioned composite fungus agent, described Brevibacillus brevis is separating obtained from the rabbit excrement; Pseudomonas fluorescens and thermoactinomyces sacchari bacterial strain are all separating obtained from bacterium slag (Lentinus Edodes fungus slag), and the preserving number of three kinds of bacterial strains is respectively CCTCC NO:M2010133, CCTCC NO:M2010135, CCTCC NO:M2010134.
In the above-mentioned composite fungus agent, the separation method of described three strain bacterial strains is as follows:
The rabbit excrement that will become thoroughly decomposed and the Lentinus Edodes fungus slag that becomes thoroughly decomposed are made bacteria suspension respectively as sample, adopt dilution-plate method, isolated strains from this bi-material, and concrete grammar is: get respectively the 10g sample, with the sterilized water dilution, make successively 10
-1, 10
-2, 10
-3, 10
-4, 10
-5, 10
-6With 10
-7The bacteria suspension of g/mL, bacteria suspension with the different concns gradient, inject respectively 1mL to culture dish, then pour in 40-50 ℃ the liquid nutrient medium (minutes three kinds: extractum carnis substratum, PDA substratum, improvement Gause I substratum), each culture dish mixes carefully that to make mixed bacterium dull and stereotyped, after culture medium solidifying, placing 45-50 ℃ of incubator to be inverted flat board cultivates, till growing bacterium colony, picking list bacterium colony, separation and purification on corresponding flat board, preservation.
After above separation test step, 27 strain bacterium have been obtained;
Again every kind of bacterial strain is put in the pig manure as single microbial inoculum, carried out fermented manure, filter out the effective microbial inoculum that is suitable for pig manure; Obtain three strain effective strains: Brevibacillus brevis, Pseudomonas fluorescens and thermoactinomyces sacchari.
Described screening method can for by monitoring every kind of bacterial strain to the index that pig manure becomes thoroughly decomposed speed and protects nitrogen, after the strict computational analysis, filter out the effective microbial inoculum that is suitable for pig manure.Obtain three strain effective strains by above-mentioned test: Brevibacillus brevis, Pseudomonas fluorescens and thermoactinomyces sacchari.
The preparation method of composite fungus agent of the present invention is as follows:
(1) slant culture: three bacterial strains that separation is obtained access respectively nutrient agar under aseptic condition, Brevibacillus brevis and Pseudomonas fluorescens are adopted the extractum carnis substratum, thermoactinomyces sacchari adopts improvement Gause I substratum, and (optimum temps is 50 ℃) cultivated 1～4 day in 45 ℃～50 ℃ incubators;
(2) first order seed is cultivated: step 1) is cultivated the bacterial classification that obtains be inoculated in respectively under aseptic condition in liquid nutrient medium corresponding to all kinds of bacterium, under 45 ℃～50 ℃ conditions (optimum temps is 50 ℃), 100～200r/min shaking table was cultivated 1～4 day, made first order seed;
(3) secondary seed is cultivated: first order seed is inoculated in respectively in the fermentor tank through 120 to 125 ℃ (optimum temps is 121 ℃) sterilization 25～30 minutes, cultivated 1～4 day 45～50 ℃ (optimum temps is 50 ℃), the bacterium number reaches 400,000,000～700,000,000 in every mL fermented liquid, makes secondary seed; Fermented liquid pH value in the described fermentor tank is 7.0-7.3, fermented liquid is comprised of water, soybean milk, starch, glucose/sucrose, calcium carbonate and iron trichloride, and the part by weight of each component is in the fermented liquid: water: the soybean of preparation soya-bean milk: starch: glucose/sucrose: calcium carbonate: iron trichloride=1000:6.5:4:1:0.1:0.01;
(4) mixing fermentation culture: the secondary seed that in proportion step (3) is made mixes, and is connected in the mixed fermentation tank that has added the wheat bran skin, and incubated at room temperature 2～4 days obtains composite fungus agent; The volume ratio of secondary seed and wheat bran skin is 1:15～1:20.
The present invention also provides the purposes of above-mentioned composite fungus agent, and this composite fungus agent is applicable to pig manure.
The concrete grammar step that composite fungus agent of the present invention is used for pig manure is as follows:
(1) in the pig manure raw material, adds straw, make that the C/N mass ratio is that 20-30:1, weight in wet base are 50%-60%, pH value 7-7.5 in the organic materials;
(2) add the ratio of 50-80mL zymocyte liquid in every 1000g organic materials, add composite fungus agent in the organic materials that obtains to step (1), mix rear heap fermentation;
When (3) compost temperature rose to 50 ℃, the ratio that adds the 20-50mL zymocyte liquid in every 1000g organic materials added composite fungus agent, mixes thoroughly; Carried out artificial turning every 3 days, until pig manure becomes thoroughly decomposed.
The present invention isolates Brevibacillus brevis, Pseudomonas fluorescens and actinomycetes thermoactinomyces sacchari three strain bacterial strains from rabbit excrement, bacterium slag, and the zymocyte liquid of the bacterial strain that separation is obtained mixes by suitable proportion, forms composite fungus agent.Composite fungus agent of the present invention has stronger capacity of decomposition, and under certain C/N, humidity, pH condition, the microorganism energy accelerated degradation organic materials in the composite fungus agent produces CO
2, H
2O, NH
3Deng gas and release energy, generate a large amount of small-molecule substances, easy decomposition small organic molecule and nutritive substance that microorganism utilizes Primary Fermentation to produce are grown, these small-molecule substances are further decomposed, become Available N, P, K compound and stable humic acid and the amino acid material that can be utilized by plant absorbing.Composite fungus agent of the present invention can accelerate the degraded of organic materials, and can reduce the nitrogen that organonitrogen is converted into gaseous form, so that in the pig manure fermenting process, can show stronger ammonia ability and the capacity of decomposition of pressing down, at the ammonia of accelerating to discharge when pig manure becomes thoroughly decomposed in a small amount.
Compared with prior art, beneficial effect of the present invention: composite fungus agent of the present invention can adapt to the middle high temperature with compost environment, and after pig manure added composite fungus agent in earlier stage, microbial inoculum can be bred in ramp, the compost organism is carried out fast degradation, and can effectively reduce the generation of ammonia.By actual compost, process relatively with the test that does not connect composite fungus agent, produce pig manure with this microbial inoculum and only need 14～18 days, shorten 1～2 times of maturity than traditional compost, can be up to 70% to the inhibiting rate of ammonia, nitrogen accumulation increases by 9%～10%.Therefore, by the effect of this composite fungus agent, pig manure can reach quick composting and reduce the ammonia volatilization, thereby reduces generation and the nitrogen loss of stench.
Description of drawings:
Fig. 1 is the variation of different treatment pH value of compost;
Fig. 2 is that different treatment is on the impact of pig manure ammonia amount;
The variation of Fig. 3 different treatment pig manure rate of emergence.
Embodiment
The present invention is described in further detail below in conjunction with test example and embodiment.But this should be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following embodiment, all technology that realizes based on content of the present invention all belong to scope of the present invention.
One, the separation of bacterial strain:
Bacterial strain among the embodiment that the present invention enumerates separates as follows and obtains:
The rabbit excrement that will become thoroughly decomposed and the Lentinus Edodes fungus slag that becomes thoroughly decomposed are made bacteria suspension respectively as sample, adopt dilution-plate method, isolated strains from this bi-material, and concrete grammar is: get respectively the 10g sample, with the sterilized water dilution, make successively 10
-1, 10
-2, 10
-3, 10
-4, 10
-5, 10
-6With 10
-7The bacteria suspension of g/mL, bacteria suspension with the different concns gradient, inject respectively 1mL to culture dish, then pour into respectively in 40-50 ℃ the different liqs substratum (liquid nutrient medium is respectively: extractum carnis substratum, PDA substratum, improvement Gause I substratum), each culture dish carefully mixes makes mixed bacterium flat board, after culture medium solidifying, placing 45-50 ℃ of incubator to be inverted flat board cultivates, till growing bacterium colony, picking list bacterium colony, separation and purification on corresponding flat board, preservation.
After above separation test step, 27 strain bacterium have been obtained.Again every kind of bacterial strain is put in the pig manure as single microbial inoculum, carried out fermented manure, by monitoring every kind of bacterial strain to the index that pig manure becomes thoroughly decomposed speed and protects nitrogen, after the strict computational analysis, filter out the effective microbial inoculum that is suitable for pig manure.Obtain three strain effective strains by above-mentioned test: Brevibacillus brevis, Pseudomonas fluorescens and thermoactinomyces sacchari.
Two, the evaluation of bacterial strain:
Three bacterial strains that screening is obtained send the evaluation by Sangon Biotech (Shanghai) Co., Ltd., wherein Brevibacillus brevis: yellowish pink, Gram-positive, shaft-like; Pseudomonas fluorescens: Gram-negative, shaft-like; Thermoactinomyces sacchari: all produce single spore on gas silk and base silk, the perfectly round shape of spore, polyhedron shape and are given birth on short handle, and the gas silk is shorter, and the very fast dissolving of mycelium stays the spore thick-layer, makes bacterium colony appearance such as bacterium.Brevibacillus brevis 16S rDNA gene order is seen the 1st sequence in the sequence table, Pseudomonas fluorescens 16S rDNA gene order is seen the 2nd sequence in the sequence table, thermoactinomyces sacchari 16S rDNA gene order is seen the 3rd sequence in the sequence table, draw three bacterial strains be respectively Brevibacillus brevis (
Brevibacillus brevis), Pseudomonas fluorescens (
Pseudomonas fluorescens) and the actinomycetes thermoactinomyces sacchari (
Thermoactinomyces sacchari).
Three, the preparation of composite fungus agent:
The preparation method that the present invention enumerates composite fungus agent is as follows:
(1) slant culture: the bacterial strain that separation is obtained accesses respectively nutrient agar under aseptic condition, Brevibacillus brevis and Pseudomonas fluorescens are adopted the extractum carnis substratum, thermoactinomyces sacchari adopts improvement Gause I substratum, cultivates 1～4 day in 45 ℃～50 ℃ incubators;
(2) first order seed is cultivated: step (1) is cultivated the bacterial classification that obtains be inoculated in respectively (bacterium employing extractum carnis substratum in liquid nutrient medium corresponding to all kinds of bacterium under aseptic condition, thermoactinomyces sacchari adopts improvement Gause I substratum,), under 45 ℃～50 ℃ conditions (optimum temps is 50 ℃), 100～200r/min shaking table was cultivated 1～4 day, made first order seed;
(3) secondary seed is cultivated: first order seed is inoculated in respectively in the fermentor tank sterilized 25～30 minutes through 120～125 ℃ (optimum temps is 121 ℃), cultivated 1～4 day at 45～50 ℃, the bacterium number reaches 400,000,000～700,000,000 in every mL fermented liquid, makes secondary seed; Fermented liquid pH value in the described fermentor tank is 7.0-7.3, fermented liquid is comprised of water, soybean milk, starch, glucose/sucrose, calcium carbonate and iron trichloride, the content of each component is in the fermented liquid: water 1000kg, 6.5kg the soya-bean milk (mass percentage concentration is 6%～8%) of soybean preparation, starch 4kg, glucose 1kg, calcium carbonate 100g, iron trichloride 10g;
(4) mixing fermentation culture: the secondary seed that in proportion step (3) is made mixes, and is connected in the mixed fermentation tank that has added the wheat bran skin, cultivates 2～4 days, obtains composite fungus agent; Fermented liquid in the described fermentor tank forms with fermented liquid in the step, and the volume ratio of fermented liquid and wheat bran skin is 1:15～1:20.
Four, the purposes of composite fungus agent.
The concrete grammar step that the composite fungus agent that the present invention enumerates is used for pig manure is as follows:
(1) add straw in the pig manure raw material, the mass ratio that makes C/N in the organic materials is that 20-30:1, weight in wet base are 50%-60%, pH value 7-7.5;
(2) add the ratio of 50-80mL zymocyte liquid in every 1000g organic materials, add composite fungus agent in the organic materials that obtains to step (1), mix rear heap fermentation;
When (3) compost temperature rose to 50 ℃, the ratio that adds the 20-50mL zymocyte liquid in every 1000g organic materials added composite fungus agent, mixes thoroughly; Carried out artificial turning every 3 days, until pig manure becomes thoroughly decomposed.
Five, specific embodiment
Embodiment 1
The composite fungus agent for the treatment of pig manure that this enforcement is enumerated, it is comprised of the component of following volume ratio: 15 parts of Brevibacillus brevis zymocyte liquids, 40 parts of Pseudomonas fluorescens zymocyte liquids, 45 parts of thermoactinomyces sacchari zymocyte liquids are prepared from according to the preparation method of above-mentioned composite fungus agent.Contain bacterium or actinomycetes about 400,000,000 in every mL zymocyte liquid.
After composite fungus agent was cultivated and to be finished, the concrete grammar that is used for pig manure by above-mentioned composite fungus agent added into pig manure and ferments.At the 2nd, 5,10,15 of fermentation reactor system, 20 days sampling analysis, per 2～3 days replacing boric acid absorption liquids.Fresh compost sample is by solid-to-liquid ratio 1:10(w.v, with dry weight basis) add a certain amount of deionized water, vibration lixiviate 1h under the speed of 150r/min, the pH value of mensuration suspension.The filtrate that obtains through filtration behind the mensuration pH is carried out the rate of emergence test.All the other samples are used for measuring full nitrogen in the laboratory natural air drying.Adopt diluted acid titration of boron acid solution to calculate the ammonia amount.
Embodiment 2
The composite fungus agent for the treatment of pig manure that this enforcement is enumerated, it is comprised of the component of following volume ratio: 40 parts of Brevibacillus brevis zymocyte liquids, 15 parts of Pseudomonas fluorescens zymocyte liquids, 45 parts of thermoactinomyces sacchari zymocyte liquids.Contain bacterium or actinomycetes about 400,000,000 in every mL zymocyte liquid.
Embodiment 3
The composite fungus agent for the treatment of pig manure that this enforcement is enumerated, it is comprised of the component of following volume ratio: 40 parts of Brevibacillus brevis zymocyte liquids, 40 parts of Pseudomonas fluorescens zymocyte liquids, 20 parts of thermoactinomyces sacchari zymocyte liquids.Contain bacterium or actinomycetes 600,000,000 in every mL zymocyte liquid.
Embodiment 4
The composite fungus agent for the treatment of pig manure that this enforcement is enumerated, it is comprised of the component of following volume ratio: 30 parts of Brevibacillus brevis zymocyte liquids, 30 parts of Pseudomonas fluorescens zymocyte liquids, 40 parts of thermoactinomyces sacchari zymocyte liquids.Contain bacterium or actinomycetes 400,000,000 in every mL zymocyte liquid.
This embodiment is most preferred embodiment.
The test-results accompanying drawing 1 of most preferred embodiment, Fig. 2, Fig. 3, table 1 and table 2.
Through test result analysis, having added composite fungus agent WZ1, the whole fermentation period of pig manure is shortened to is 13～16 days, and compared with the control, the ammonia rate that presses down that composite fungus agent is processed reaches more than 70%, the nitrogen increment rate reaches 9%～10%, illustrate that composite fungus agent WZ1 has effectively improved compost quality and greatly shortens pig manure and become thoroughly decomposed the time, has saved cost.
The compound WZ1 of table 1. is to the inhibition of ammonia
The nitrogen accumulation effect of the compound WZ1 of table 2.
| Process | Full nitrogen (%) before the compost | Full nitrogen (%) behind the compost | Full nitrogen accretion (%) | The nitrogen increment rate |
| Contrast | 2.134 | 1.928 d | -0.206 | -0.096 |
| Brevibacillus brevis | 2.134 | 2.297 b | 0.163 | 0.076 |
| Thermoactinomyces sacchari | 2.134 | 2.279 b | 0.145 | 0.068 |
| Pseudomonas fluorescens | 2.134 | 2.245 c | 0.111 | 0.052 |
| Compound WZ1 | 2.134 | 2.344 a | 0.210 | 0.098 |
Sequence table
＜110〉Sichuan Agricultural University
＜120 〉
A kind of composite fungus agent and its preparation method and application
<160>?3
<210>?1
<211>?1495
<212>?DNA
＜213 〉
Brevibacillus brevis(
Brevibacillus brevis)
<400>?1
agagtttgat?catggctcag?gacgaacgct?ggcggcgtgc?ctaatacatg?caagtcgagc?60
gaggatcttc?ggatcctagc?ggcggacggg?tgagtaacac?gtaggcaacc?tgcctctcag?120
accgggataa?catagggaaa?cttatgctaa?taccggatag?gtttttggat?cgcatgatct?180
gaaaaggaaa?gatggcttct?gctatcactg?ggagatgggc?ctgcggcgca?ttagctagtt?240
ggtggggtaa?cggcctacca?aggcgacgat?gcgtagccga?cctgagaggg?tgaccggcca?300
cactgggact?gagacacggc?ccagactcct?acgggaggca?gcagtaggga?attttccaca?360
atggacgaaa?gtctgatgga?gcaacgccgc?gtgaacgatg?aaggtcttcg?gattgtaaag?420
ttctgttgtc?agggacgaat?aagtaccgtt?cgaacagggc?ggtaccttga?cggtacctga?480
cgagaaagcc?acggctaact?acgtgccagc?agccgcggta?atacgtaggt?ggcaagcgtt?540
gtccggaatt?attgggcgta?aagcgcgcgc?aggcggctat?gtaagtctgg?tgttaaagcc?600
cggggctcaa?ccccggttcg?catcggaaac?tgtgtagctt?gagtgcagaa?gaggaaagcg?660
gtattccacg?tgtagcggtg?aaatgcgtag?agatgtggag?gaacaccagt?ggcgaaggcg?720
gctttctggt?ctgtaactga?cgctgaggcg?cgaaagcgtg?gggagcaaac?aggattagat?780
accctggtag?tccacgccgt?aaacgatgag?tgctaggtgt?tgggggtttc?aataccctca?840
gtgccgcagc?taacgcaata?agcactccgc?ctggggagta?cgctcgcaag?agtgaaactc?900
aaaggaattg?acgggggccc?gcacaagcgg?tggagcatgt?ggtttaattc?gaagcaacgc?960
gaagaacctt?accaggtctt?gacatcccgc?tgaccgctct?ggagacagag?cttcccttcg?1020
gggcagcggt?gacaggtggt?gcatggttgt?cgtcagctcg?tgtcgtgaga?tgttgggtta?1080
agtcccgcaa?cgagcgcaac?ccttatcttt?agttgccagc?attcagttgg?gcactctaga?1140
gagactgccg?tcgacaagac?ggaggaaggc?ggggatgacg?tcaaatcatc?atgcccctta?1200
tgacctgggc?tacacacgtg?ctacaatggt?tggtacaacg?ggatgctacc?tcgcgagagg?1260
acgccaatct?cttaaaacca?atctcagttc?ggattgtagg?ctgcaactcg?cctacatgaa?1320
gtcggaatcg?ctagtaatcg?cggatcagca?tgccgcggtg?aatacgttcc?cgggccttgt?1380
acacaccgcc?cgtcacacca?cgggagtttg?caacacccga?agtcggtgag?gtaaccgtaa?1440
ggagccagcc?gccgaaggtg?gggtagatga?ctggggtgaa?gtcgtaacaa?ggtaa?1495
<210>?2
<211>?1498
<212>?DNA
＜213〉Pseudomonas fluorescens (
Pseudomonas fluorescens)
<400>?2
agagtttgat?catggctcag?attgaacgct?ggcggcaggc?ctaacacatg?caagtcgagc?60
ggtagagaga?agcttgcttc?tcttgagagc?ggcgaacggg?tgagtaaagc?ctaggaatct?120
gcctggtagt?gggggataac?gttcggaaac?ggacgctaat?accgcatacg?tcctacggga?180
gaaagcaggg?gaccttcggg?ccttgcgcta?tcagatgagc?ctaggtcgga?ttagctagtt?240
ggtgaggtaa?tggctcacca?aggcgacgat?ccgtaactgg?tctgagagga?tgatcagtca?300
cactggaact?gagacacggt?ccagactcct?acgggaggca?gcagtgggga?atattggaca?360
atgggcgaaa?gcctgatcca?gccatgccgc?gtgtgtgaag?aaggtcttcg?gattgtaaag?420
cactttaagt?tgggaggaag?ggcattaacc?taatacgtta?gtgttttgac?gttaccgaca?480
gaataagcac?cggctaactc?tgtgccagca?gccgcggtaa?tacagagggt?gcaagcgtta?540
atcggaatta?ctgggcgtaa?agcgcgcgta?ggtggtttgt?taagttggat?gtgaaatccc?600
cgggctcaac?ctgggaactg?cattcaaaac?tgactgacta?gagtatggta?gagggtggtg?660
gaatttcctg?tgtagcggtg?aaatgcgtag?atataggaag?gaacaccagt?ggcgaaagcg?720
accacctgga?ctaatactga?cactgaggtg?cgaaagcgtg?gggagcaaac?aggattagat?780
accctggtag?tccacgccgt?aaacgatgtc?agctagccgt?tgggagcctt?gagctcttag?840
tggcgcagct?aacgcattaa?gttgaccgcc?tggggagtac?ggccgcaagg?ttaaaactca?900
aatgaattga?cgggggcccg?cacaagcggt?ggagcatgtg?gtttaattcg?aagcaacgcg?960
aagaacctta?ccaggccttg?acatccaatg?aactttctag?agatagattg?gtgccttcgg?1020
gaacattgag?acaggtgctg?catggctgtc?gtcagctcgt?gtcgtgagat?gttgggttaa?1080
gtcccgtaac?gagcgcaacc?cttgtcctta?gttaccagca?cgtaatggtg?ggcactctaa?1140
ggagactgcc?ggtgacaaac?cggaggaagg?tggggatgac?gtcaagtcat?catggccctt?1200
acggcctggg?ctacacacgt?gctacaatgg?tcggtacaga?gggttgccaa?accgcgaggt?1260
ggagctaatc?ccacaaaacc?gatcgtagtc?cggatcgcag?tctgcaactc?gactgcgtga?1320
agtcggaatc?gctagtaatc?gcgaatcaga?atgtcgcggt?gaatacgttc?ccgggccttg?1380
tacacaccgc?ccgtcacacc?atgggagtgg?gttgcaccag?aagtagctag?tctaaccttc?1440
gggaggacgg?ttaccacggt?gtgattcatg?actggggtga?agtcgtaaca?aggtaacc?1498
<210>?3
<211>?1466
<212>?DNA
＜213〉and the actinomycetes thermoactinomyces sacchari (
Thermoactinomyces sacchari)
<400>?3
agagtttgat?cctggctcag?gatgaacgct?ggtggcatgc?ctaatacatg?caagtcgtgc?60
gggggtctcc?ggaccctagc?ggcgaacggg?tgagtaacac?gtgggcaacc?tgcccgtaag?120
actgggataa?ctccgggaaa?ccggtgctaa?taccagataa?tctctttctc?cgcatggaga?180
aagagtaaaa?ggcttcggcc?acttacggat?gggcccgcgg?cgcattagct?agttggtgag?240
gtaacggctc?accaaggcga?cgatgcgtag?ccgacctgag?agggtgatcg?gccacactgg?300
gactgagaca?cggcccagac?tcctacggga?ggcagcagta?gggaattttc?cgcaatgggc?360
gaaagcctga?cggagcaatg?ccgcgtgagt?gatgacggcc?ttcgggttgt?aaaactctgt?420
tctgaaggaa?gaagttctga?cggtacttca?ggagaaagcc?ccggctaact?acgtgccagc?480
agccgcggta?atacgtaggg?ggcgagcgtt?atccggaatt?attgggcgta?aagcgcgcgc?540
aggcggtcat?ttaagtcagg?tgtgaaaggc?tgcggctcaa?ccgcagagcg?gcacctgaaa?600
ctggatgact?tgagtgcagg?agaggagagc?ggaattcccg?gtgtagcggt?ggaatgcgta?660
gagatcggga?ggaacaccag?tggcgaaggc?ggctctctgg?cctgtttctg?acgctgaggc?720
gcgaaagcgt?ggggagcaaa?caggattaga?taccctggta?gtccacgccg?taaacgatga?780
gtgctaggtg?tcggggggtc?ctaccctcgg?tgccgaaagg?taacccatta?agcactccgc?840
ctggggagta?cggccgcaag?gctgaaactc?aaaggaattg?acgggggccc?gcacaagcgg?900
tggagcatgt?ggtttaattc?gaagcaacgc?gaagaacctt?accagggctt?gacatcccgc?960
tgaccgctcc?agagatggag?cttcccttcg?gggcagcggt?gacaggtggt?gcatggttgt?1020
cgtcagctcg?tgtcgtgaga?tgttgggttc?agtcccgcaa?cgagcgcaac?ccttatcgct?1080
agttgccagc?attcagttgg?gcactctagc?gagacagccg?gtgaaagccg?gaggaaggtg?1140
gggatgacgt?caaatcatca?tgccccttat?gtcctgggct?acacacgtgc?tacaatggcc?1200
ggtacaatgg?gtcgccaacc?cgcgaggggg?agctaatccc?acaaaaccgg?tctcagttcg?1260
gatcgcaggc?tgcaactcgc?ctgcgtgaag?ccggaatcgc?tagtaatcgc?ggatcagcat?1320
gccgcggtga?atacgttccc?gggccttgta?cacaccgccc?gtcacaccac?gagagtttgc?1380
aacacccgaa?gtcggtgagg?taaccttttg?gaaccagccg?ccgaaggtgg?ggcagatgat?1440
tggggtgaag?tcgtaacaag?gtaacc?1466
Claims (5)
1. composite fungus agent, it is characterized in that it is comprised of the component of following volume ratio: 10～40 parts of Brevibacillus brevis zymocyte liquids, 10～40 parts of Pseudomonas fluorescens zymocyte liquids, 15～45 parts of thermoactinomyces sacchari zymocyte liquids, in the zymocyte liquid of above-mentioned each component in every ml fermented liquid the bacterium number be 4 hundred million-7 hundred million, Brevibacillus brevis is separating obtained from the rabbit excrement; Pseudomonas fluorescens and thermoactinomyces sacchari bacterial strain are all separating obtained from the Lentinus Edodes fungus slag, the preserving number of three kinds of bacterial strains be respectively Brevibacillus brevis (
Brevibacillus brevis): CCTCC NO:M2010133, Pseudomonas fluorescens (
Pseudomonas fluorescens): CCTCC NO:M2010135, thermoactinomyces sacchari (
Thermoactinomyces sacchari): CCTCC NO:M2010134.
2. composite fungus agent according to claim 1 is characterized in that each zymocyte liquid volume ratio is: 15～35 parts of Brevibacillus brevis zymocyte liquids, 15～35 parts of Pseudomonas fluorescens zymocyte liquids, 20～40 parts of thermoactinomyces saccharis.
3. composite fungus agent according to claim 2 is characterized in that each zymocyte liquid volume ratio is: 30 parts of Brevibacillus brevis zymocyte liquids, 30 parts of Pseudomonas fluorescens zymocyte liquids, 40 parts of thermoactinomyces sacchari zymocyte liquids.
4. the purposes of the described composite fungus agent of claim 1-3 any one is characterized in that: described composite fungus agent is used for pig manure.
5. the purposes of composite fungus agent according to claim 4 is characterized in that: the concrete grammar step that described composite fungus agent is used for pig manure is as follows:
(1) in the pig manure raw material, adds straw, make that the C/N mass ratio is that 20-30:1, weight in wet base are 50%-60%, pH value 7-7.5 in the organic materials;
(2) add the ratio of 50-80mL zymocyte liquid in every l000g organic materials, add composite fungus agent in the organic materials that obtains to step (1), mix rear heap fermentation;
When (3) compost temperature rose to 50 ℃, the ratio that adds the 20-50mL zymocyte liquid in every 1000g organic materials added composite fungus agent, mixes thoroughly; Carried out artificial turning every 3 days, until pig manure becomes thoroughly decomposed.
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