CN108624539A - A kind of stalk and cow dung superhigh temperature decomposing agent and its preparation and application - Google Patents

A kind of stalk and cow dung superhigh temperature decomposing agent and its preparation and application Download PDF

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CN108624539A
CN108624539A CN201810586225.8A CN201810586225A CN108624539A CN 108624539 A CN108624539 A CN 108624539A CN 201810586225 A CN201810586225 A CN 201810586225A CN 108624539 A CN108624539 A CN 108624539A
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microbial inoculum
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cow dung
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CN108624539B (en
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王松艳
刘辉
周天惠
王灵敏
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Hebei Rimidori Technology Co ltd
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Abstract

The invention discloses a kind of stalk and cow dung superhigh temperature decomposing agent and its preparation and application, belong to decomposing agent field.The decomposing agent is obtained by being dried after stalk, cow dung and decomposed composite bacteria agent mixed fermentation, wherein decomposed composite bacteria agent includes 1 10 parts of thermophilic bacteria agent, 16 parts of bacillus licheniformis agent, 16 parts of aspergillus oryzae microbial inoculum, 16 parts of lactobacillus plantarum microbial inoculum, 16 parts of thermoactinomyces sacchari microbial inoculum, 16 parts of brevibacterium frigoritolerans microbial inoculum.The present invention is directed to existing composite bacteria fermentation problem：Compost maturity plays temperature slowly, and fermentation temperature is low, and fermentation time is long, decomposed to be not thorough, compost quality difference etc. provides a kind of compost maturity and plays that temperature is fast, and heating is rapid, and fermentation temperature is high, fermentation time is short, it is decomposed more thoroughly, the higher decomposing agent of quality and its compost method and application.

Description

A kind of stalk and cow dung superhigh temperature decomposing agent and its preparation and application
Technical field
The invention belongs to decomposing agent field, more particularly to a kind of stalk and cow dung superhigh temperature decomposing agent and its prepares and answer With.
Background technology
With the development of agricultural and animal husbandry, agricultural crop straw and animal wastes are treated as big problem, once processing is not Well the survival and development of the mankind can be caused to seriously endanger, very big burden can be brought to environment.But from the angle of resource reutilization Analysis, stalk and animal wastes ingredient are more complicated, such as carbohydrate, lipid, protein, nitrogen, phosphorus, potassium, magnesium, sulphur organic matter And inorganic salts, it can turn waste into wealth by processing, microbial fermentation is current most common method.Due to agricultural crop straw and ox The main component of excrement is cellulose, difficult decomposed substance, and the heating of traditional natural fermentating method is slow, decomposed to be not thorough, and ferments Period is long, and not only efficiency is low in this way, but also secondary pollution can be caused to environment, therefore, all the time, there is this side both at home and abroad The correlative study in face improves the speed and quality of fermentation using microbial bacterial agent, but what is mostly used greatly is all mesophilic micoorganism, Warm speed of having fermented is slow, when the tolerable temperature microorganism that fermentation temperature is more than mesophilic micoorganism existence will be all dead, in this way It may result in being not thorough for fermentation.Patent at present about microorganism-decomposing microbial inoculum is relatively more：
Chinese patent CN103614326B discloses a kind of microbial inoculum that the fermentation highest temperature reaches 50 DEG C -60 DEG C, and microbial inoculum heat is fine Clostridium, stearothermophilus bacillus, Taiwan vacation Xanthomonas campestris, soil bacillus brevis and bacillus licheniformis proportioning be (3- 5)：(1-2)：(1-2)：(1-2)：(1-2) reduces nitrogen loss although the present invention saves the energy, and fermentation temperature is relatively low, Certain influence is had on rotten degree.
Chinese patent CN103642701B discloses a kind of microbial inoculum that the fermentation highest temperature reaches 50 DEG C -60 DEG C, microbial inoculum proportioning Long 5-7 parts of seta shell freeze-drying lactobacillus powder, 4-6 parts of trichodermaharzianum freeze-drying lactobacillus powder, circle ferment hair shell freeze-drying lactobacillus powder 3-5 Part, 3-5 parts of Trichoderma viride freeze-drying lactobacillus powder and 2-3 parts of geotrichum candidum freeze-drying lactobacillus powder, bacteria fermentation temperature of the present invention compare It is low, can have a certain impact to the rotten degree of fermentate, and cannot by fermentate worm's ovum and harmful microorganism all kill Extremely, compost quality is influenced.
Chinese patent CN102660479B discloses a kind of microbial inoculum that the fermentation highest temperature reaches 75 DEG C, and microbial inoculum is by 14 plants of bacterium groups At microbial bacterial agent of the invention can make product total viable count more, be conducive to improve material rotten degree, but utilize the present invention Microbial inoculum decomposed period it is long, probably need 36 days terminate it is decomposed, greatly reduce decomposed efficiency.
Chinese patent CN1460664A discloses a kind of microbial inoculum that the fermentation highest temperature reaches 70 DEG C -90 DEG C, and microbial inoculum is by 4-8 plants Thermophilic ligocellulose degradation bacterium composition, the present invention promote compost maturity by the synergistic effect of high temperature bacterium and mesophile, The decomposed quality of product is improved, but the one time fermentation stage is more preferably sent out to make the microorganism in composite bacteria agent 1 keep activity Degradation function is waved, maximum temperature must be controlled at 75 DEG C hereinafter, this standard is difficult to meet in process of production.
Patent microbial inoculum type disclosed above is more, and the fermentation temperature that can reach is different, is nearly all mesophile, rises Warm slow, fermentation temperature is low to cause fermentate degradation to be not thorough, and miscellaneous bacteria, worm's ovum and virus are more in fermentate, due to fermentation temperature Spend it is relatively low its cannot be made lethal, therefore fermentate be applied to fertilizer can become seedling, wood, soil infection pest and disease damage main pollution Source can all damage seedling, wood, soil applied to orchard and field.Selection is suitble to microbial inoculum that fermentate temperature is made quickly to increase, Reach the decomposed requirement of high-quality, it is rapid decomposed, it is most important to shorten fermentation time.
Invention content
Problem solved by the invention is to be directed to existing composite bacteria fermentation problem：Compost maturity plays temperature slowly, fermentation temperature Low, fermentation time is long, decomposed to be not thorough, and compost quality is poor, it is difficult to reach the quality requirement etc. that people use fermented manure, provide A kind of compost maturity plays that temperature is fast, and heating is rapid, and fermentation temperature is high, and fermentation time is short, it is decomposed thoroughly, the higher decomposing agent of quality and Its compost method and application.
In order to achieve the above object, the present invention uses following experimental programs：
A kind of stalk and cow dung superhigh temperature decomposing agent, by being dried after stalk, cow dung and decomposed composite bacteria agent mixed fermentation It obtains, it is characterised in that：The decomposed composite bacteria agent is made of following parts by weight raw material：Thermophilic bacteria agent 1-10 parts, lichens 1-6 parts of gemma bacillus agent, 1-6 parts of aspergillus oryzae microbial inoculum, 1-6 parts of lactobacillus plantarum microbial inoculum, 1-6 parts of thermoactinomyces sacchari microbial inoculum, 1-6 parts of brevibacterium frigoritolerans microbial inoculum.
Preferably, the decomposed composite bacteria agent is mainly made of following parts by weight raw material：
Thermophilic bacteria agent 3-8 parts, 2-5 parts of bacillus licheniformis agent, 2-5 parts of aspergillus oryzae microbial inoculum, lactobacillus plantarum microbial inoculum 2-5 parts, 2-5 parts of thermoactinomyces sacchari microbial inoculum, 2-5 parts of brevibacterium frigoritolerans microbial inoculum；
It is highly preferred that the decomposed composite bacteria agent, is mainly made of following parts by weight raw material：
6 parts of thermophilic bacteria agent, 4 parts of bacillus licheniformis agent, 4 parts of aspergillus oryzae microbial inoculum, 4 parts of lactobacillus plantarum microbial inoculum are sweet Sugarcane orchid wishes 4 parts of Salmonella microbial inoculum, 4 parts of brevibacterium frigoritolerans microbial inoculum；
Preferably, the Thermophilic Bacteria is specially Thermophilic Bacteria (Thermaerobacter composti) GW, and deposit number is CGMCC No.15721；
Preferably, the preparation method of the thermophilic bacteria agent, includes the following steps：Thermophilic Bacteria is inoculated in high-temperature strain In solid slope culture medium A, after 60-90 DEG C of culture 18-22h, thalline is eluted, elution bacterium solution is obtained, adjusts bacterium solution Cell concentration is 4-6 × 1011Cfu/mL is inoculated according to inoculum concentration 50-150mL/kg culture mediums in solid medium B, mixing Stirring, 60-90 DEG C of stationary culture 16-20h, 1-3 times/day of moisturizing obtain thermophilic bacteria agent, microbial inoculum content after drying, crushing ≥6×1011cfu/g。
It is highly preferred that after the fermented culture of Thermophilic Bacteria, dry 0.5-1.5d, uses pulverizer under the conditions of 70-85 DEG C It crushes, crosses 30-50 mesh and sieve up to high temperature bacteria agent.
It is highly preferred that the thermophilic bacteria agent content is 6-7 × 1011cfu/g。
Further, the sub- solid medium A groups of the high-temperature strain become：Yeast extract 2-3g/L, bacto peptone 2-3g/L, glucose 2-3g/L, PIPES 6-8g/L, plant gel 8-12g/L, agar powder 8-12g/L, surplus is water, is adjusted PH value 7.2-7.4,121 DEG C of sterilizing 30min.
Further, the solid medium B groups become：100-120 parts of rice husk, 100-110 parts of wheat bran, corn flour 100- 120 parts, 100-115 parts of beancake powder, 100-110 parts of glucose, 2-3 parts of calcium sulfate, 2-4 parts of calcium oxide, dipotassium hydrogen phosphate 1-3 Part, 0.5-1.2 parts of magnesium sulfate, according to solid material total amount and water weight ratio：5-6:Water, 121 DEG C of sterilizings are added into solid material by 4-5 45min。
Preferably, the bacillus licheniformis is bacillus licheniformis CICC 10037, bacillus licheniformis CICC 10084, any one or a few in bacillus licheniformis CICC 10085, bacillus licheniformis CICC 10092.
It is highly preferred that the bacillus licheniformis is bacillus licheniformis CICC 10037.
Preferably, the preparation method of the bacillus licheniformis agent, includes the following steps：Bacillus licheniformis is inoculated with In NA seed solid mediums C, 30-40 DEG C of stationary culture 1-2d elutes thalline, obtains elution bacterium solution, adjusts bacterium A concentration of 3-5 × 10 of liquid10Cfu/mL is inoculated according to inoculum concentration 50-120mL/kg culture mediums in solid medium D, and stirring is mixed It closes, 30-40 DEG C of stationary culture 1-2d, after drying, crushing, obtains bacillus licheniformis agent, microbial inoculum content >=4 × 1010cfu/g；
It is highly preferred that after the fermented culture of bacillus licheniformis, dry 3-4d under the conditions of 30-40 DEG C, with crushing Machine crushes, and crosses 30-50 mesh and sieves up to bacillus licheniformis agent.
It is highly preferred that the bacillus licheniformis agent content is 4-5 × 1010cfu/g；
Further, the group of the NA seeds solid medium C becomes：Peptone 10-12g/L, powdered beef 3-4g/L, chlorine Change sodium 10-12g/L, agar 18-20g/L, surplus is water, and adjustment pH value is 7.0-7.2,121 DEG C of sterilizing 30min；
Further, the group of the solid medium D becomes：250-275 parts of rice husk, 500-750 parts of wheat bran, corn flour 375-625 parts, 200-275 parts of beancake powder, 225-250 parts of starch, 250-275 parts of laterite, 50-62 parts of glucose, calcium sulfate 12- 15 parts, 15-20 parts of calcium oxide, 5-10 parts of dipotassium hydrogen phosphate, 5-6 parts of magnesium sulfate is 7- according to solid material total amount and water weight ratio 15:Water mixing, 121 DEG C of sterilizing 45min are added into solid material by 2-5；
Preferably, the aspergillus oryzae is aspergillus oryzae CICC 2001, aspergillus oryzae CICC 2011, aspergillus oryzae CICC2016, rice Any one or a few in aspergillus CICC 2022.
It is highly preferred that the aspergillus oryzae is aspergillus oryzae CICC 2001；
Preferably, the preparation method of the aspergillus oryzae microbial inoculum, includes the following steps：Aspergillus oryzae strain is inoculated in PDA to consolidate In body culture medium, 25-30 DEG C of stationary culture 5-6d elutes thalline, obtains elution bacterium solution, and adjustment bacterial concentration is 2-4 ×1011Cfu/mL is inoculated according to inoculum concentration 50-120mL/kg culture mediums in solid medium B, 25-30 DEG C of stationary culture 5- 6d obtains aspergillus oryzae viable bacteria microbial inoculum, microbial inoculum content >=8 × 10 after drying, crushing10cfu/g。
It is highly preferred that after the fermented culture of aspergillus oryzae, dry 3-5d under the conditions of 25-30 DEG C, with pulverizer into Row crushes, and crosses 30-50 mesh and sieves up to bacillus licheniformis agent.
It is highly preferred that the aspergillus oryzae microbial inoculum microbial inoculum content is 8-10 × 1010cfu/g。
Further, the group of the PDA solid mediums becomes：Potato 200-250g/L, glucose 20-25g/L, agar 20-22g/L, surplus are water, and pH is naturally, 121 DEG C of sterilizing 30min.
Preferably, the lactobacillus plantarum is lactobacillus plantarum CGMCC1.2437.
Preferably, the preparation method of the lactobacillus plantarum microbial inoculum, includes the following steps：Lactobacillus plantarum is inoculated in kind In sub- solid medium C, 30-40 DEG C of stationary culture 2-3d elutes thalline, obtains elution bacterium solution, adjusts bacterial concentration For 4-5 × 1010Cfu/mL is inoculated in solid medium F, 30-40 by bacterium solution according to inoculum concentration 50-120mL/kg culture mediums DEG C stationary culture 2-3d, obtains lactobacillus plantarum microbial inoculum, microbial inoculum content >=8 × 10 after drying and crushing10cfu/g。
It is highly preferred that after the fermented culture of lactobacillus plantarum, dry 2-3d, uses pulverizer under the conditions of 30-40 DEG C It crushes, crosses 30-50 mesh and sieve up to lactobacillus plantarum microbial inoculum.
It is highly preferred that the lactobacillus plantarum microbial inoculum content is 8-9 × 1010cfu/g。
Further, the preparation method of the solid medium F is：300-500 parts of beancake powder, 300-500 parts of rice husk, cotton 100-200 parts of the dregs of rice, 2-5 parts of peptone, 1-5 parts of glucose, 0.1-0.5 parts of magnesium sulfate, according to solid material and water quality ratio 7-10: 4-6 adds water into solid material, uniformly mixes, and 121 DEG C of sterilizing 45min to obtain the final product.
Preferably, the thermoactinomyces sacchari is specially thermoactinomyces sacchari (Laceyella sacchari) CP, preservation Number is CGMCC No.13928；
Preferably, the preparation method of the thermoactinomyces sacchari microbial inoculum, includes the following steps：Thermoactinomyces sacchari is inoculated with In NA seed solid mediums C, 50-70 DEG C of stationary culture 1-2d elutes thalline, obtains elution bacterium solution, adjusts bacterium A concentration of 5-5.5 × 10 of liquid11Cfu/mL is inoculated in solid medium A by bacterium solution according to inoculum concentration 50-120mL/kg culture mediums In, 50-70 DEG C of stationary culture 4-5d obtains thermoactinomyces sacchari viable bacteria microbial inoculum after drying, crushing, and microbial inoculum content >=7 × 1011cfu/g；
It is highly preferred that after the fermented culture of thermoactinomyces sacchari, dry 2-4d under the conditions of 50-70 DEG C, with crushing Machine crushes, and crosses 30-50 mesh and sieves up to thermoactinomyces sacchari microbial inoculum.
It is highly preferred that the thermoactinomyces sacchari microbial inoculum content is 7-8 × 1011cfu/g；
Preferably, the brevibacterium frigoritolerans are specially brevibacterium frigoritolerans (Brevibacterium frigoritolerans) NH, deposit number are CGMCC No.15722；
Preferably, the preparation method of the brevibacterium frigoritolerans microbial inoculum, includes the following steps：Brevibacterium frigoritolerans are inoculated in NA In seed solid medium C, 8-10 DEG C of stationary culture 2-3d elutes thalline, obtains elution bacterium solution, adjusts bacterial concentration For 8-9 × 1010Cfu/mL is inoculated in solid medium G, 8-10 DEG C by bacterium solution according to inoculum concentration 50-110mL/kg culture mediums Stationary culture 3-4d obtains brevibacterium frigoritolerans microbial inoculum, microbial inoculum content >=3 × 10 after drying, crushing11cfu/g；
It is highly preferred that after the fermented culture of brevibacterium frigoritolerans, dry 7-8d under the conditions of 8-10 DEG C, with pulverizer into Row crushes, and crosses 30-50 mesh and sieves up to thermoactinomyces sacchari microbial inoculum.
It is highly preferred that the brevibacterium frigoritolerans microbial inoculum content is 3-5 × 1011cfu/g；
Further, the group of the solid medium G becomes：Wheat bran 600-620g/L, rice husk 180-200g/L, peanut cake Powder 80-100g/L, dregs of beans 100-110g/L, magnesium sulfate 0.5-0.8g/L, ammonium sulfate 3-5g/L, surplus are water, 121 DEG C of sterilizings 45min to obtain the final product.
Another object of the present invention is to provide the preparation method of a kind of above-mentioned stalk and cow dung superhigh temperature decomposing agent, including as follows Step：
(1) microbial inoculum mixes：According to above-mentioned parts by weight by thermophilic bacteria agent, bacillus licheniformis agent, aspergillus oryzae microbial inoculum, Lactobacillus plantarum microbial inoculum, thermoactinomyces sacchari microbial inoculum, brevibacterium frigoritolerans microbial inoculum, are mixed, and are crossed 30-40 mesh sieve, are obtained decomposed Composite bacteria agent；
(2) by fresh straw and cow dung according to mass ratio 10-20：1-5 is mixed, and obtains hybrid reactor body, the hybrid reactor body Water content 55%-65%；
(3) inoculum concentration of the decomposed composite bacteria agent 1-2 ‰ in mass ratio is inoculated into hybrid reactor body, is mixed, Decomposed processing is carried out, compost 10-15d obtains stalk and cow dung superhigh temperature decomposing agent.
Another object of the present invention is to provide a kind of above-mentioned stalk and cow dung superhigh temperature decomposing agent in fruits and vegetables field of planting Purposes.
Preferably, the purposes of the stalk and cow dung superhigh temperature decomposing agent in rapeseed cultivation.
It is highly preferred that the purposes of the stalk and cow dung superhigh temperature decomposing agent in rapeseed cultivation, steps are as follows：By high temperature Decomposing agent is added according to the ratio of mass percent 1% in soil, after carrying out sowing field planting later, after rape seed 50 days at It is ripe.
Strain source：
1, Thermophilic Bacteria (Thermaerobacter composti) GW is to be detached from the fermented manure of Qinhuangdao, is pure Change, screening obtains.The bacterial strain is preserved in China Committee for Culture Collection of Microorganisms on May 2nd, 2018 (abbreviation CGMCC, address are common micro-organisms center：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Chinese Academy of Sciences microorganism Research institute, postcode：100101) it is Thermophilic Bacteria that, deposit number, which is CGMCC No.15721 Classification And Nomenclatures, (Thermaerobacter composti)。
The Thermophilic Bacteria (Thermaerobacter composti) can grow within the scope of 50-100 DEG C, the most suitable growth temperature Degree is 80 DEG C；
High temperature bacterium of the Thermophilic Bacteria (Thermaerobacter composti) in the different pH values that pH value is 3-11 It is coated with high temperature bacterium tablet after fluid nutrient medium culture and is cultivated with 80 DEG C in liquid high temperature bacterium culture medium test tube, can well give birth to Long, detection bacterium work content is 4-6 × 1011Cfu/g, two experiments all prove that the bacterial strain can be resistant to the ranging from 3- of pH value 11, than broad, with stronger degrees.
2, the bacillus licheniformis CICC 10037, bacillus licheniformis CICC 10084, bacillus licheniformis CICC 10085, bacillus licheniformis CICC 10092 is bought in Chinese industrial Microbiological Culture Collection administrative center.Selected by the present invention Bacillus licheniformis, at 37 DEG C~50 DEG C, it can exist growth temperature with spore form, to resist severe ring Border.Meanwhile resistant activity material can be generated, and there is unique biology take oxygen mechanism, the growth of pathogenic bacteria can be inhibited numerous It grows.
3, the aspergillus oryzae CICC 2001, aspergillus oryzae CICC 2011, aspergillus oryzae CICC 2016, aspergillus oryzae CICC 2022 It buys in Chinese industrial Microbiological Culture Collection administrative center.Aspergillus oryzae selected by the present invention, 28 DEG C of growth temperature, is removed It produces outside protease, can also produce amylase, carbohydrase, cellulase etc., cellulose that can be effectively in degrading straw and cow dung forms sediment Powder, protein and polysaccharose substance, to be effectively converted into fertilizer.
4, the lactobacillus plantarum is lactobacillus plantarum CGMCC1.2437, is bought in China General Microbiological culture presevation Administrative center.Lactobacillus plantarum selected by the present invention, growth temperature are 30 DEG C -35 DEG C, are a kind of natural antiseptic agents, can have Effect reduces ammonia nitrogen and nitrite yield in digest process.
5, the thermoactinomyces sacchari CP is to detach, purify from the decomposed object of Hebei province Qinhuangdao cow dung high temperature, screening It obtains, deposit number is CGMCC No.13928, and Classification And Nomenclature is thermoactinomyces sacchari (Laceyella sacchari), should Bacterium is in Publication No. CN106916772A, and entitled " one plant of acid and alkali-resistance, the sugarcane orchid of ethanol-tolerant quick composting vinasse are uncommon It is disclosed in the patent of invention of Salmonella and its application ".
The thermoactinomyces sacchari CP of the present invention, growth temperature are 30 DEG C~75 DEG C, have high temperature resistant, ethanol-tolerant, acidproof Alkali characteristic, can generate a variety of degrading enzymes such as protease, lipase, the ranging from 3-12 of pH tolerant, and tolerance alcoholic strength range exists 1-13% can not only effectively improve adaptability of the fertilizer for different soda acid soil, but also for alcohol in fertilizer fermentation process The generation of fermentation process has significant tolerance.
6, the brevibacterium frigoritolerans are specially brevibacterium frigoritolerans (Brevibacterium frigoritolerans) NH, real It is separating obtained to test room, it is general which has been preserved in China Committee for Culture Collection of Microorganisms on May 2nd, 2018 (abbreviation CGMCC, address are at logical microorganism center：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Chinese Academy of Sciences microorganism is ground Study carefully institute, postcode：100101), deposit number is CGMCC No.15722, and Classification And Nomenclature is brevibacterium frigoritolerans (Brevibacterium frigoritolerans)。
Advantageous effect：
1, the stalk for preparing of the present invention and cow dung superhigh temperature decomposing agent according to different regions temperature difference is larger and stalk and Cow dung composed structure and nutritional ingredient characteristic will be resistant to low temperature, medium temperature, high temperature, the bacterial strain science compounding for decomposing various composition, profit With mutually being acted synergistically between different decomposing microbial inoculums to cellulose, lignin, protein, starch, the fat etc. in stalk and cow dung Ingredient carries out fully degraded, effectively increases degradation capability of the decomposing agent to stalk and cow dung.
2, brevibacterium frigoritolerans and Thermophilic Bacteria have significant low temperature resistant and high-temperature stability.
Brevibacterium frigoritolerans coordinate Thermophilic Bacteria with its high- and low-temperature resistance and the characteristic for the fermentation that is rapidly heated, and high temperature can be made decomposed Agent in the lower rapid fermentation calefaction in place of northern temperature, substantially reduce fermentation into the megathermal period time (1d reaches 60 DEG C, 3d reaches 98 DEG C), Thermophilic Bacteria can survive under hyperthermal environments, avoid all dead fermentate degradations of the excessively high bacterial strain of temperature Incomplete situation, material rotten degree greatly improve, and maintain the time length (85-98 DEG C of maintenance 3d) of hot fermentation, shorten Fermentation period (only 15d), effectively increases the yield and quality of organic fertilizer, it is therefore prevented that stalk and decomposed be not thorough of cow dung are made At secondary pollution and human and material resources, financial resources, waste of time, be stalk and the decomposed making high quality organic fertilizer of cow dung high temperature Provide a new way.
3, Thermophilic Bacteria has significant bacterial strain vigor, and fermentation can be rapidly completed, and has saved the time cost of microbial inoculum preparation, It is inoculated in the sub- solid slope culture medium A of high-temperature strain, after cultivating 18-22h at 60-90 DEG C, thalline can be paved with inclined-plane, It is inoculated in solid medium B, in 60-90 DEG C of stationary culture 16-20h, you can complete fermentation.
4, bacillus licheniformis (bacillus licheniformis CICC 10037, bacillus licheniformis CICC disclosed by the invention 10084, bacillus licheniformis CICC 10085, bacillus licheniformis CICC 10092), can exist with spore form, from And resist rugged environment.Resistant activity material can be generated, and there is unique biology take oxygen mechanism, pathogenic bacteria can be inhibited Growth and breeding.
5, aspergillus oryzae disclosed by the invention can also produce amylase, carbohydrase, cellulase etc. in addition to producing protease, can be with Cellulose, starch, protein and polysaccharose substance in effective degrading straw and cow dung.
6, lactobacillus plantarum disclosed by the invention is a kind of natural antiseptic agent, significantly reduce in digest process ammonia nitrogen and Nitrite yield.
7, thermoactinomyces sacchari disclosed by the invention have high temperature resistant, ethanol-tolerant, acid and alkali-resistance characteristic, can generate protease, A variety of degrading enzymes such as lipase, the ranging from 3-12 of pH tolerant, tolerance alcoholic strength range is in 1-13%.
Description of the drawings
Fig. 1 be during stalk and cow dung compost Bu Tong under decomposed processing temperature variation；
Fig. 2 be during stalk and cow dung compost Bu Tong under decomposed processing pH value variation；
Fig. 3 be during stalk and cow dung compost Bu Tong under decomposed processing the content of organic matter variation；
Fig. 4 be during stalk and cow dung compost Bu Tong under decomposed processing total nitrogen content variation；
During Fig. 5 stalks and cow dung compost Bu Tong under decomposed processing water content variation.
Specific implementation mode
The preparation method of 1 stalk of embodiment and cow dung superhigh temperature decomposing agent
(1) microbial inoculum mixes：By 6 parts of thermophilic bacteria agent, 4 parts of bacillus licheniformis agent, 4 parts of aspergillus oryzae microbial inoculum, plant breast 4 parts of 4 parts of bacillus microbial inoculum, 4 parts of thermoactinomyces sacchari microbial inoculum, brevibacterium frigoritolerans microbial inoculum mixing, cross 40 mesh sieve, obtain decomposed compound bacteria Agent；
(2) by fresh straw and cow dung according to mass ratio 5:1 mixing, obtains hybrid reactor body, the hybrid reactor body it is aqueous Measure 55%-65%；
(3) inoculum concentration of the decomposed composite bacteria agent in mass ratio 1 ‰ is inoculated into hybrid reactor body, is mixed, into The decomposed processing of row, under conditions of initial compost temperature is 10 DEG C, compost 13d obtains stalk and cow dung superhigh temperature decomposing agent.
Wherein, Thermophilic Bacteria bacterial preparation process includes the following steps：Thermophilic Bacteria slant strains are inoculated in height with transfer needle In the warm sub- solid medium A of strain, then 80 DEG C of culture 18h elute thalline with sterile water, adjustment cell concentration is 4 ×1011Cfu/mL draws 100mL bacterium solutions and is inoculated into 1kg solid mediums B, and 80 DEG C of culture 16h are respectively mended in every morning, afternoon Water ventilation is primary, and dry 1d, is crushed with pulverizer under the conditions of 80 DEG C, crosses 40 mesh and sieves up to high temperature bacteria agent, wherein high Warm bacterium viable bacteria content is 6 × 1011cfu/g。
The sub- solid medium A preparation methods of high-temperature strain are：Yeast extract 2.5g, bacto peptone 2.5g, Portugal Grape sugar 2.5g, PIPES 7.5g, plant gel 10g, agar powder 10g, sterile water 1L are uniformly mixed, and are fully dissolved, and pH value is adjusted 7.2-7.4 (solid state N aOH adjusts PH), is dispensed into after boiling in seed, every bottle of 80-100mL, 121 DEG C of sterilizing 30min are put into tiltedly Face to obtain the final product.
The solid medium B preparation methods are：Rice husk 100g, wheat bran 100g, corn flour 100g, beancake powder 100g, Portugal Grape sugar 100g, calcium sulfate 2.5g, calcium oxide 3g, dipotassium hydrogen phosphate 1.5g, magnesium sulfate 1g, water 400mL are uniformly mixed, and 121 DEG C go out Bacterium 45min to obtain the final product.
The bacillus licheniformis is bacillus licheniformis CICC 10037；
The preparation method of the bacillus licheniformis agent, includes the following steps：Bacillus licheniformis slant strains are used Transfer needle is inoculated in NA seed solid mediums C, then thalline is eluted with sterile water, adjusts bacterium by 40 DEG C of culture 1d Bulk concentration is 3 × 1010Cfu/mL draws 100mL bacterium solutions and is inoculated into 1kg solid mediums D, 40 DEG C of culture 2d, in 40 DEG C of items Dry 3d, is crushed with pulverizer under part, is crossed 40 mesh and is sieved up to bacillus licheniformis agent, bacillus licheniformis viable bacteria content It is 4 × 1010cfu/g；
The preparation method of the NA seeds solid medium C is：Peptone 10g, powdered beef 3g, sodium chloride 10g, agar 20g, distilled water 1L are uniformly mixed, and are fully dissolved, and adjustment pH value is 7.0, is dispensed into after boiling in seed bottle, every bottle of 80- 100mL, 121 DEG C of sterilizing 30min, is put into inclined-plane to obtain the final product；
The preparation method of the solid medium D is：Rice husk 260g, wheat bran 600g, corn flour 500g, beancake powder 230g, Starch 230g, laterite 260g, glucose 55g, calcium sulfate 13g, calcium oxide 18g, dipotassium hydrogen phosphate 8g, magnesium sulfate 5g, water 1360mL is uniformly mixed, and 121 DEG C of sterilizing 45min to obtain the final product；
The aspergillus oryzae is aspergillus oryzae CICC 2001；
The preparation method of the aspergillus oryzae microbial inoculum, includes the following steps：Aspergillus oryzae slant strains are inoculated in transfer needle In PDA seed solid mediums E, 28 DEG C culture 5d, then thalline is eluted with sterile water, adjustment cell concentration be 2 × 1011Cfu/mL draws 100mL bacterium solutions and is inoculated into 1kg solid mediums B, 28 DEG C of culture 5d, dry 3d under the conditions of 28 DEG C, It is crushed with pulverizer, crosses 40 mesh and sieve up to bacillus licheniformis agent, aspergillus oryzae viable bacteria content is 8 × 1010cfu/g。
The preparation method of the PDA seeds solid medium E is：Potato 200g, is cut into small pieces, and appropriate amounts of sterilized water is added to boil 20min, filtered through gauze take filtrate, and glucose 20g, agar 20g, it is 1L to mend distilled water to culture volume, is uniformly mixed, fully Dissolving, is dispensed into seed bottle, every bottle of 80-100mL after boiling, and 121 DEG C of sterilizing 30min are put into inclined-plane to obtain the final product；
The lactobacillus plantarum is lactobacillus plantarum CGMCC1.2437；
The preparation method of the lactobacillus plantarum microbial inoculum, includes the following steps：Lactobacillus plantarum slant strains are inoculated with Needle is inoculated in seed solid medium C, then thalline is eluted with sterile water, adjusts cell concentration by 35 DEG C of culture 2d It is 4 × 1010Cfu/mL draws 100mL bacterium solutions and is inoculated into 1kg solid mediums F, and 35 DEG C of culture 2d are done under the conditions of 35 DEG C Dry 2d, is crushed with pulverizer, is crossed 40 mesh and is sieved up to lactobacillus plantarum microbial inoculum, and lactobacillus plantarum viable bacteria content is 8 × 1010cfu/g。
The preparation method of the solid medium F is：Beancake powder 400g, rice husk 350g, cotton dregs 120g, peptone 3g, Portugal Grape sugar 2g, magnesium sulfate 0.2g, water 547mL are uniformly mixed, and 121 DEG C of sterilizing 45min to obtain the final product.
The thermoactinomyces sacchari is thermoactinomyces sacchari CGMCC No.13928；
The preparation method of the thermoactinomyces sacchari microbial inoculum, includes the following steps：Thermoactinomyces sacchari slant strains are used Transfer needle is inoculated in NA seed solid mediums C, 60 DEG C of culture 1d, and thalline is scraped with sterile water elution, adjusts thalline A concentration of 5 × 1011Cfu/mL draws 100mL bacterium solutions and is inoculated into 1kg solid mediums A, 60 DEG C of culture 4d, in 60 DEG C of conditions Lower dry 2d, is crushed with pulverizer, is crossed 40 mesh and is sieved up to thermoactinomyces sacchari CP microbial inoculums, wherein thermoactinomyces sacchari viable bacteria Content 7 × 1011cfu/g；
The preparation method of the brevibacterium frigoritolerans microbial inoculum, includes the following steps：Brevibacterium frigoritolerans slant strains are inoculated with Needle is inoculated in NA seed solid mediums C, 10 DEG C of culture 3d, and thalline is scraped with sterile water elution, adjusts cell concentration It is 8 × 1010Cfu/mL draws 100mL bacterium solutions and is inoculated into 1kg solid mediums G, and 10 DEG C of culture 3d are done under the conditions of 10 DEG C Dry 7d, is crushed with pulverizer, is crossed 40 mesh and is sieved up to thermoactinomyces sacchari microbial inoculum, wherein brevibacterium frigoritolerans viable bacteria content is 3 ×1011cfu/g；
The preparation method of the solid medium G is：Wheat bran 600g, rice husk 200g, groundnut meal 100g, dregs of beans 100g, Magnesium sulfate 0.5g, ammonium sulfate 5g, water 1L are uniformly mixed, and 121 DEG C of sterilizing 45min to obtain the final product.
The preparation method of 2 stalk of embodiment and cow dung superhigh temperature decomposing agent
(1) microbial inoculum mixes：By 3 parts of thermophilic bacteria agent, 2 parts of bacillus licheniformis agent, 2 parts of aspergillus oryzae microbial inoculum, plant breast 2 parts of bacillus microbial inoculum, 2 parts of thermoactinomyces sacchari microbial inoculum, the mixing of 2 parts of brevibacterium frigoritolerans microbial inoculum cross 30 mesh sieve to get to decomposed compound Microbial inoculum；
(2) by fresh straw and cow dung according to mass ratio 11:6 mixing, obtain hybrid reactor body, the hybrid reactor body it is aqueous Measure 60%-65%；
(3) inoculum concentration of the decomposed composite bacteria agent in mass ratio 1.5 ‰ is inoculated into hybrid reactor body, is mixed, Under conditions of initial compost temperature is 10 DEG C, decomposed processing is carried out, compost 10d obtains stalk and cow dung superhigh temperature decomposing agent.
Wherein, Thermophilic Bacteria bacterial preparation process includes the following steps：Thermophilic Bacteria slant strains are inoculated in height with transfer needle In the warm sub- solid medium A of strain, 75 DEG C culture 4d, then thalline is eluted with sterile water, adjustment cell concentration be 5 × 1011Cfu/mL draws 125mL bacterium solutions and is inoculated into 1kg solid mediums B, 75 DEG C of culture 5d, every morning, each moisturizing in afternoon Ventilation is primary, and dry 1d, is crushed with pulverizer under the conditions of 75 DEG C, crosses 40 mesh and sieves up to high temperature bacteria agent, high temperature Bacterium viable bacteria content is 6.5 × 1011cfu/g。
The preparation method of the brevibacterium frigoritolerans microbial inoculum, includes the following steps：Brevibacterium frigoritolerans slant strains are inoculated with Needle is inoculated in NA seed solid mediums C, 10 DEG C of culture 3d, and thalline is scraped with sterile water elution, adjusts cell concentration It is 8 × 1010Cfu/mL draws 100mL bacterium solutions and is inoculated into 1kg solid mediums G, and 10 DEG C of culture 3d are done under the conditions of 10 DEG C Dry 7d, is crushed with pulverizer, is crossed 40 mesh and is sieved up to thermoactinomyces sacchari microbial inoculum, wherein brevibacterium frigoritolerans viable bacteria content is 3 ×1011cfu/g；
The bacillus licheniformis is bacillus licheniformis CICC 10084；
The aspergillus oryzae is aspergillus oryzae CICC 2011；
The lactobacillus plantarum is lactobacillus plantarum CGMCC1.2437；
The thermoactinomyces sacchari is thermoactinomyces sacchari CGMCC No.13928；
The preparation of the bacillus licheniformis agent, aspergillus oryzae microbial inoculum, lactobacillus plantarum microbial inoculum, thermoactinomyces sacchari microbial inoculum Method and bacterium live content with embodiment 1；
In preparation process, seed solid medium A, solid medium B, seed solid medium C, solid medium D, kind Sub- solid medium E, solid medium F, the preparation method is the same as that of Example 1 by solid medium G.
The preparation method of 3 stalk of embodiment and cow dung superhigh temperature decomposing agent
(1) microbial inoculum mixes：By 8 parts of thermophilic bacteria agent, 5 parts of bacillus licheniformis agent, 5 parts of aspergillus oryzae microbial inoculum, plant breast 5 parts of 5 parts of bacillus microbial inoculum, 5 parts of thermoactinomyces sacchari microbial inoculum, brevibacterium frigoritolerans microbial inoculum mixing, cross 40 mesh sieve, obtain decomposed compound bacteria Agent；
(2) by fresh straw and cow dung according to mass ratio 12:5 mixing, obtain hybrid reactor body, the hybrid reactor body it is aqueous Measure 55%-65%；
(3) inoculum concentration of the decomposed composite bacteria agent 1-2 ‰ in mass ratio is inoculated into hybrid reactor body, is mixed, Under conditions of initial compost temperature is 11 DEG C, decomposed processing is carried out, compost 12d obtains stalk and cow dung superhigh temperature decomposing agent.
The Thermophilic Bacteria bacterial preparation process, includes the following steps：Thermophilic Bacteria slant strains are inoculated in height with transfer needle In the warm sub- solid medium A of strain, then 82 DEG C of culture 19h elute thalline with sterile water, adjustment cell concentration is 6 ×1011Cfu/mL draws 100mL bacterium solutions and is inoculated into 1.2kg solid mediums B, 82 DEG C of culture 19h, and every morning, afternoon are each Moisturizing ventilation is primary, and dry 1d, is crushed with pulverizer under the conditions of 82 DEG C, crosses 40 mesh and sieves up to high temperature bacteria agent, wherein High temperature bacterium viable bacteria content is 7 × 1011cfu/g。
The preparation method of the brevibacterium frigoritolerans microbial inoculum, includes the following steps：Brevibacterium frigoritolerans slant strains are inoculated with Needle is inoculated in NA seed solid mediums C, 10 DEG C of culture 3d, and thalline is scraped with sterile water elution, adjusts cell concentration It is 9 × 1010Cfu/mL draws 100mL bacterium solutions and is inoculated into 1kg solid mediums G, and 10 DEG C of culture 3d are done under the conditions of 10 DEG C Dry 7d, is crushed with pulverizer, is crossed 40 mesh and is sieved up to thermoactinomyces sacchari microbial inoculum, wherein brevibacterium frigoritolerans viable bacteria content is 4.5×1011cfu/g；
The bacillus licheniformis is bacillus licheniformis CICC10085；
The aspergillus oryzae is aspergillus oryzae CICC2016；
The lactobacillus plantarum is lactobacillus plantarum CGMCC1.2437；
The thermoactinomyces sacchari is thermoactinomyces sacchari CGMCC No.13928；
The preparation of the bacillus licheniformis agent, aspergillus oryzae microbial inoculum, lactobacillus plantarum microbial inoculum, thermoactinomyces sacchari microbial inoculum Method and bacterium live content with embodiment 1；
In preparation process, seed solid medium A, solid medium B, seed solid medium C, solid medium D, kind Sub- solid medium E, solid medium F, the preparation method is the same as that of Example 1 by solid medium G.
The preparation method of 4 stalk of embodiment and cow dung superhigh temperature decomposing agent
(1) microbial inoculum mixes：By 1 part of thermophilic bacteria agent, 1 part of bacillus licheniformis agent, 1 part of aspergillus oryzae microbial inoculum, plant breast 1 part of 1 part of bacillus microbial inoculum, 1 part of thermoactinomyces sacchari microbial inoculum, brevibacterium frigoritolerans microbial inoculum mixing, cross 40 mesh sieve, obtain decomposed compound bacteria Agent；
(2) by fresh straw and cow dung according to mass ratio 6:1 mixing, obtains hybrid reactor body, the hybrid reactor body it is aqueous Measure 55%-65%；
(3) inoculum concentration of the decomposed composite bacteria agent in mass ratio 2 ‰ is inoculated into hybrid reactor body, is mixed, Under conditions of initial compost temperature is 12 DEG C, decomposed processing is carried out, compost 10d obtains stalk and cow dung superhigh temperature decomposing agent.
The Thermophilic Bacteria bacterial preparation process, includes the following steps：Thermophilic Bacteria slant strains are inoculated in height with transfer needle In the warm sub- solid medium A of strain, then 78 DEG C of culture 18h elute thalline with sterile water, adjustment cell concentration is 7 ×1011Cfu/mL draws 100mL bacterium solutions and is inoculated into 1.0kg solid mediums B, 78 DEG C of culture 16h, and every morning, afternoon are each Moisturizing ventilation is primary, and dry 1d, is crushed with pulverizer under the conditions of 78 DEG C, crosses 40 mesh and sieves up to high temperature bacteria agent, wherein High temperature bacterium viable bacteria content is 7.2 × 1011cfu/g。
The preparation method of the brevibacterium frigoritolerans microbial inoculum, includes the following steps：Brevibacterium frigoritolerans slant strains are inoculated with Needle is inoculated in NA seed solid mediums C, 10 DEG C of culture 3d, and thalline is scraped with sterile water elution, adjusts cell concentration It is 8 × 1010Cfu/mL draws 100mL bacterium solutions and is inoculated into 1kg solid mediums G, and 10 DEG C of culture 3d are done under the conditions of 10 DEG C Dry 7d, is crushed with pulverizer, is crossed 40 mesh and is sieved up to thermoactinomyces sacchari microbial inoculum, wherein brevibacterium frigoritolerans viable bacteria content is 3 ×1011cfu/g；
The bacillus licheniformis is bacillus licheniformis CICC10092；
The aspergillus oryzae is aspergillus oryzae CICC2022；
The lactobacillus plantarum is lactobacillus plantarum CGMCC1.2437；
The thermoactinomyces sacchari is thermoactinomyces sacchari CGMCC No.13928；
The preparation of the bacillus licheniformis agent, aspergillus oryzae microbial inoculum, lactobacillus plantarum microbial inoculum, thermoactinomyces sacchari microbial inoculum Method and bacterium live content with embodiment 1；
In preparation process, seed solid medium A, solid medium B, seed solid medium C, solid medium D, kind Sub- solid medium E, solid medium F, the preparation method is the same as that of Example 1 by solid medium G.
The preparation method of 5 stalk of embodiment and cow dung superhigh temperature decomposing agent
(1) microbial inoculum mixes：By 8 parts of thermophilic bacteria agent, 5 parts of bacillus licheniformis agent, 5 parts of aspergillus oryzae microbial inoculum, plant breast 5 parts of 5 parts of bacillus microbial inoculum, 5 parts of thermoactinomyces sacchari microbial inoculum, brevibacterium frigoritolerans microbial inoculum mixing, cross 40 mesh sieve, obtain decomposed compound bacteria Agent；
(2) by fresh straw and cow dung according to mass ratio 18：5 mixing, obtain hybrid reactor body, the hybrid reactor body it is aqueous Measure 55%-65%；
(3) inoculum concentration of the decomposed composite bacteria agent in mass ratio 1.5 ‰ is inoculated into hybrid reactor body, is mixed, Under conditions of initial compost temperature is 10 DEG C, decomposed processing is carried out, compost 10d obtains stalk and cow dung superhigh temperature decomposing agent.
The Thermophilic Bacteria bacterial preparation process, includes the following steps：Thermophilic Bacteria slant strains are inoculated in height with transfer needle In the warm sub- solid medium A of strain, then 80 DEG C of culture 18h elute thalline with sterile water, adjustment cell concentration is 5 ×1011Cfu/mL draws 100mL bacterium solutions and is inoculated into 1.2kg solid mediums B, 80 DEG C of culture 17h, and every morning, afternoon are each Moisturizing ventilation is primary, and dry 1d, is crushed with pulverizer under the conditions of 80 DEG C, crosses 40 mesh and sieves up to high temperature bacteria agent, wherein High temperature bacterium viable bacteria content is 5.6 × 1011cfu/g。
The preparation method of the brevibacterium frigoritolerans microbial inoculum, includes the following steps：Brevibacterium frigoritolerans slant strains are inoculated with Needle is inoculated in NA seed solid mediums C, 10 DEG C of culture 3d, and thalline is scraped with sterile water elution, adjusts cell concentration It is 8 × 1010Cfu/mL draws 100mL bacterium solutions and is inoculated into 1kg solid mediums G, and 10 DEG C of culture 3d are done under the conditions of 10 DEG C Dry 7d, is crushed with pulverizer, is crossed 40 mesh and is sieved up to thermoactinomyces sacchari microbial inoculum, wherein brevibacterium frigoritolerans viable bacteria content is 3 ×1011cfu/g；
The bacillus licheniformis is bacillus licheniformis CICC10037 and bacillus licheniformis CICC10084；
The aspergillus oryzae is aspergillus oryzae CICC2001, aspergillus oryzae CICC2011 and aspergillus oryzae CICC2016；
The lactobacillus plantarum is lactobacillus plantarum CGMCC1.2437；
The thermoactinomyces sacchari is thermoactinomyces sacchari CGMCC No.13928；
The preparation of the bacillus licheniformis agent, aspergillus oryzae microbial inoculum, lactobacillus plantarum microbial inoculum, thermoactinomyces sacchari microbial inoculum Method and bacterium live content with embodiment 1；
In preparation process, seed solid medium A, solid medium B, seed solid medium C, solid medium D, kind Sub- solid medium E, solid medium F, the preparation method is the same as that of Example 1 by solid medium G；
The bacillus licheniformis CICC10037 and bacillus licheniformis CICC10084 microbial inoculums in mass ratio 2:1 is uniformly mixed It closes；
The aspergillus oryzae CICC2001, aspergillus oryzae CICC2011 and aspergillus oryzae CICC2016 microbial inoculums in mass ratio 1:1:1 Even mixing.
6 Thermophilic Bacteria pH value tolerance test of embodiment
100mL high temperature bacteria liquid culture mediums are accessed from the appropriate thalline of Thermophilic Bacteria slant strains picking, 80 DEG C, 200r/min shakes Bed culture 12h obtains seed liquor, and it is respectively 1.5,2,2.5,3,3.5,4 that seed liquor is accessed pH with 1 ‰ (volume ratio) inoculum concentrations, In 4.5,5,5.5,6,6.5,7,7.5,8,8.5,9,9.5,10,10.5,11,11.5,12 high temperature bacteria liquid culture medium, 80 DEG C, it is coated with NA tablets after 200r/min shaking table culture cultures 16h, observes growing state；Simultaneously by above-mentioned seed liquor with 1 ‰ inoculations The high temperature bacteria liquid culture medium test tube of amount access same pH gradient, observes turbidity by 80 DEG C, 200r/min shaking table culture 16h.
The group of its high temperature bacteria liquid culture medium becomes：Yeast extract 2.5g, peptone 2.5g, glucose 2.5g, PIPES 7.5g, pH value 7.2-7.4 (solid state N aOH is adjusted), water 1L
Test result：Thermophilic Bacteria 80 DEG C of culture 16h, coating NA tablets in the high temperature bacteria liquid culture medium of pH value 3-11 is equal Energy well-grown, 3 bacterium of pH value live content as 1-2 × 1010Cfu/ml, 11 bacterium of pH value live content as 2-4 × 1010Cfu/ml, pH value 5-9 detects bacterium and lives content as 5-6 × 1011cfu/ml；After 80 DEG C of NA fluid nutrient mediums test tube, 200r/min shaking table cultures 16h Well-grown, turbidity is apparent, and pH value 5-9 detection bacterium live content as 4.5-5.5 × 1011Cfu/ml, two experiments prove thermophilic Hot bacterium can tolerate pH ranges in 3-11, and acid-base value tolerance is than wide.
7 Thermophilic Bacteria temperature tolerance of embodiment is tested with genetic stability
100mL high temperature bacteria liquid culture mediums (ingredient is with embodiment 6) are accessed from the appropriate thalline of Thermophilic Bacteria slant strains picking, 80 DEG C, 200r/min shaking table cultures 12h obtains seed liquor, is coated on the tablet of solid medium H, (its middle plateform culture used Base be the sub- solid slope culture medium A of high-temperature strain) be respectively placed in 40 DEG C, 45 DEG C, 50 DEG C, 55 DEG C, 60 DEG C, 65 DEG C, 70 DEG C, 75 DEG C, 80 DEG C, 85 DEG C, 90 DEG C, 95 DEG C, 100 DEG C, 105 DEG C, 110 DEG C culture 18h, observe growing state.Simultaneously by seed liquor with 1 ‰ (volume ratio) inoculum concentrations access high temperature bacteria liquid culture medium test tube, are respectively placed under above-mentioned identical temperature gradient, 200r/ Min shaking table culture 16h observe turbidity.
Wherein, solid medium H ingredients are：Yeast extract 2.5g, peptone 2.5g, glucose 2.5g, PIPES 7.5g, PH value 7.2-7.4 (solid state N aOH is adjusted), 1% plant gel, 1% agar, water 1L.
Test result：After Thermophilic Bacteria Liquid Culture 12h, on spread plate, in 50 DEG C -100 DEG C equal energy well-growns, inspection Survey is cultivated under the conditions of 100 DEG C, and viable bacteria content is 5.5-6.0 × 1011cfu/ml；In high temperature bacteria liquid culture medium test tube 200r/ Well-grown after min shaking table cultures 16h, clearly, detection bacterium lives content as 5.5-6.5 × 10 to turbidity11Cfu/ml, two It is 50 DEG C -100 DEG C that item experiment, which proves that Thermophilic Bacteria can tolerate growth temperature, with higher temperature tolerance.
Secondary culture is carried out to Thermophilic Bacteria in this way, through passage ten times, the work of the bacterium per a generation is detected and is not less than 1010Cfu/mL, and growthform and characteristic are constant, illustrate that the bacterial strain can stablize heredity, and can well grow.
The acid and alkali-resistance of 8 brevibacterium frigoritolerans of embodiment measures
Experimental method：(1) with PH count adjust every bottle of culture medium pH value, be tuned into respectively pH value size be 3,4,5,6,7,8,9, 10,11,12, sterilizing is down flat plate.
(2) brevibacterium frigoritolerans slant strains are inoculated in transfer needle in NA seed solid mediums, 10 DEG C of culture 4d, and Thalline is scraped with sterile water elution, dilution spread is on the different culture medium of 10 kinds of pH values, 1-2 days observation bacterial strains of 28 degree of cultures Upgrowth situation.
Wherein, NA culture mediums group becomes：Peptone 10g, powdered beef 3g, Nacl 5g, PH 7.2,1% agar, water 1L.
Experimental result：By the observation to bacterium colony upgrowth situation on different PH tablet, find brevibacterium frigoritolerans in pH value All fine for what is grown on 5,6,7,8,9,10,11,12 culture medium, spawn activity is 3-5 × 1010A/ml.PH value is 3,4 Solid medium do not solidified to tablet when tablet, therefore again configuration pH value be 3,4 fluid nutrient medium shake bacterium cultivate bacterial strain, Bacterial strain vigor is 2-5 × 1010A/ml.From the experimental results, brevibacterium frigoritolerans are resistant to acid-base value range and are at least 3-12, May more extensively, spawn activities of the pH within the scope of 3-12 is 0.5-2 × 1010A/ml.
The salt resistant test of 9 brevibacterium frigoritolerans of embodiment
Experimental method：(1) add Nacl according to 2%, 5%, 10%, 15% ratio respectively in each conical flask, sterilize, Tablet.
(2) brevibacterium frigoritolerans slant strains are inoculated in transfer needle in NA seed solid mediums, 10 DEG C of culture 4d, and Thalline is scraped with sterile water elution, on the culture medium of dilution spread to four kinds of different salinities, every plant of bacterium is on each culture medium Activate 4 tablets, 1-2 days observation strain growth situations of 28 degree of cultures.
Experimental result：Brevibacterium frigoritolerans are grown all fine on the culture medium of salinity 2%, 5%, 10%, 15%, strain Vigor reaches 0.5-4 × 1010A/ml.
Embodiment 10 detects brevibacterium frigoritolerans tolerable temperature range
Experimental method：Brevibacterium frigoritolerans slant strains are inoculated in transfer needle in NA seed solid mediums, 10 DEG C of trainings Support 3d, and thalline scraped with sterile water elution, on dilution spread to NA tablets, be put into respectively 4 DEG C, 7 DEG C, 25 DEG C, 30 DEG C, 35 DEG C, 40 DEG C, 50 DEG C, 60 DEG C, 70 DEG C culture 1-2 days observation as a result, each temperature 4 tablets of every plant of bacterium as parallel.
Experimental result：
The spawn activity of 1 different temperatures of table
" +++ " indicates to grow vigorous, spawn activity 2-4 × 1010A/ml, " ++ " indicate well-grown, spawn activity 0.5- 2×1010A/ml.
From the point of view of experimental result, brevibacterium frigoritolerans are cultivated in 4 DEG C, 7 DEG C, 25 DEG C, 30 DEG C, 35 DEG C, 40 DEG C, 50 DEG C can Growth very well, but do not grown in 60 DEG C and 70 DEG C cultures.Thus may determine that the temperature range of brevibacterium frigoritolerans growth is at least It, may more extensively at 4 DEG C~50 DEG C.
Embodiment 11 detects brevibacterium frigoritolerans solution protein capability
2.1.3 culture medium
Experimental method：Configuration solution protein culture medium, sterilizing are down flat plate, connect brevibacterium frigoritolerans point after culture medium solidification Onto solution albumen tablet, so 6 tablets of inoculation, 28 degree are cultivated 3 days, observation experiment result.Wherein：Solve protein culture medium：Ox Meat soaks powder 3g, sodium chloride 5g, casein 2g, agar powder 20g, distilled water 1L.The a small amount of 2% sodium hydroxide dissolving profit of casein Wet stirring is added appropriate distilled water and boils dissolving, then constant volume, 7.6,121 DEG C of sterilizing 30min of pH value.
Experimental result：By observation experiment as a result, it has been found that：The circle for having solution albumen on solution albumen tablet, illustrates brevibacterium frigoritolerans With the characteristic for having significant solution albumen, albumen loop diameter is 3.2cm.
Embodiment 12 detects brevibacterium frigoritolerans solution starch performance
Experimental method：Starch culture-medium will be solved, sterilizes, is down flat plate, will be connected to brevibacterium frigoritolerans point after culture medium solidification It solves on starch tablet, is inoculated with 6 tablets, 28 degree are cultivated 5 days, tablet iodine staining, observation experiment result.Wherein：Solve starch Culture medium group becomes：Soluble starch 10g, peptone 10g, powdered beef 3g, sodium chloride 5g, distilled water 1L, agar 20g, pH value 7.2。
Experimental result：By observation experiment as a result, it has been found that：The circle for having on starch tablet and solving starch is solved, starch loop diameter is 1.4cm illustrates that brevibacterium frigoritolerans have the characteristic of solution starch.
Embodiment 13 detects brevibacterium frigoritolerans solution fat performance.
Experimental method：The fatty culture medium of configuration solution, sterilizing are down flat plate, are inoculated with brevibacterium frigoritolerans after culture medium solidification Onto the fatty tablet of solution, be afraid of that tablet is seriously polluted, so 6 tablets of inoculation, 28 degree are cultivated 5 days, observation experiment result.Wherein solve Fatty culture medium：Peptone 10g, sodium chloride 5g, calcium chloride 0.1g, pH value 7.4, distilled water 1L, agar powder 20g, 121 DEG C of sterilizings 30min is cooled to 40-50 DEG C, is separately added into Tween40, Tween60, Tween80 for individually sterilizing to final concentration 1%, shakes up It is down flat plate.
Experimental result：By observation experiment as a result, it has been found that：The circle for having solution fatty on the fatty tablet of solution, fatty loop diameter are 3cm illustrates that brevibacterium frigoritolerans have the characteristic of significant solution fat.
13 stalk of embodiment and cow dung superhigh temperature decomposing microbial inoculum carry out stalk and the decomposed experiment of cow dung compost
1. stalk and cow dung compost digest process
It is with the inoculum concentration access pH of mass ratio 1.5 ‰ by stalk and cow dung decomposing microbial inoculum prepared by the embodiment of the present invention 1 5.7, moisture content is in 60% heap body, mixes thoroughly, and 1 meter of heap body heap top width, 1.8 meters of heap bottom width, heap is 1-1.2 meters high, heap length 200 Rice.Compost initial temperature is 10 DEG C, primary with turning vehicle turning when temperature is more than 60 DEG C, later primary per turning in 1-2 days.Often It is secondary to detect temperature from five different locations of compost, a time point detection is chosen daily, it is primary (herein every turning in 3-5 days Reason is experimental group L).
It is control that two raw materials and the identical compost maturity processing of scale, which are arranged, is respectively defined as CK1 and CK2, CK1 To be inoculated with commercially available decomposing microbial inoculum, CK2 is not to be inoculated with decomposing microbial inoculum, and test operation step is identical as experimental group L as method.
The heap body of wherein experimental group L and control group CK1 and CK2 are by fresh stalk and cow dung according to mass ratio 16：5 is mixed It closes, obtains hybrid reactor body, the water content 60% of the hybrid reactor body.
2. the test result that experimental group L and control group CK1, CK2 are compared
2.1 temperature
It can be seen from attached drawing 1 under conditions of initial compost temperature is 10 DEG C, the compost heating rate of experimental group L is most Soon, and compost 1d can reach 60 DEG C, and 3d reaches 98 DEG C of maximum temperature and in 88-95 DEG C of lasting 4d, can reach compost without Evilization standard, fermenting the 15th day, it is decomposed to complete, and whole process is decomposed to be reached 7-10 days at 70 DEG C or more；Control group CK1 compost 5d reaches 60 DEG C, 8d reaches 75 DEG C of maximum temperature and only maintains 1d, and the maximum temperature that experimental group L reaches is 23 DEG C higher than control group CK1, control Group CK1 completes decomposed after 43d, and experimental group L completes decomposed Time transfer receiver and is advanced by 28d according to a group CK1；Control group CK2 is Reach within 12 days 58 DEG C of the highest temperature, completion in 55 days is decomposed, and experimental group L completes decomposed Time transfer receiver and is advanced by 40d according to a group CK2.On State the result shows that, inoculation decomposing agent can accelerate the degradation of organic matter in stalk and cow dung heap body, relative to commercially available stalk and cow dung Decomposing agent, the warm temperature of stalk and cow dung superhigh temperature decomposing agent compost maturity prepared by the addition present invention is low, accommodative ability of environment By force, fermentation efficiency is high, heating rate is fast, maximum temperature is high, maintains that time megathermal period is long, compost maturity fermentation period is short etc. significantly Superiority.
2.2PH value
PH is respectively at the end of control group CK1, the control group CK2 it can be seen from attached drawing 2 and experimental group L compost fermentations 7.6,7.8,7.3, on the whole, inoculation decomposing agent heap body PH is increased, but experimental group is lower compared with the control group for experimental group PH value can reduce the volatilization of ammonia in heap body, and then reduce the loss of nitrogen.
2.3 the content of organic matter
It can be seen from attached drawing 3 at the end of compost maturity control group CK1, control group CK2 and experimental group L organic matter Content is respectively 57.46%, 62.38% and 46.27%, reduce 33.97% respectively than the initial content of organic matter 86.43%, 27.05%, 43.16%；From content of organic matter variation it is found that inoculation decomposing microbial inoculum is larger to the degradation of compost organic matter, And the degradation efficiency of the inoculation stalk and cow dung superhigh temperature decomposing agent of the invention prepared is significantly higher than commercially available stalk and cow dung is rotten Ripe dose.
2.4 total nitrogen content
At the end of compost maturity it can be seen from attached drawing 4, the total nitrogen content of experimental group L is 2.78%, control group CK1's Total nitrogen content is 1.62%, and the total nitrogen content of control group CK2 is 1.28%.Three processing are compared, and the heap body for being inoculated with decomposing agent is complete Nitrogen content is apparently higher than control group CK1, and be inoculated with stalk and the cow dung superhigh temperature commercially available stalk of decomposing agent ratio prepared by the present invention and The total nitrogen content of cow dung high temperature decomposing agent is slightly higher.
2.5 water content
At the end of compost maturity it can be seen from attached drawing 5, the water content of experimental group L is 20.5%, and control group CK1's contains Water is 26.8%, and the water content of control group CK2 is 28.9%, and the water content for being inoculated with superhigh temperature decomposing microbial inoculum of the present invention is obviously low In the water content for being inoculated with commercially available decomposing microbial inoculum.It can be seen that being added to decomposing microbial inoculum of the present invention, the maximum temperature of fermentation reaches 98 DEG C, it is the maximum temperature having not found so far, and the time for entering high temperature is shorter, maintains the time of maximum temperature It is long, to keep the ratio of moisture evaporation very fast.
As can be seen from Table 2, after decomposed, the nitrogen, phosphorus, the potassium content (%) that each handle.
Nitrogen, phosphorus, potassium content (%) after table 2 is decomposed
Note：P1-P5 is five parallel test groups
As shown in Table 2, provide that N, P, K total amount must not be less than 5%, organic matter in organic fertilizer professional standard NY525-2012 Content must not be less than 45% and pH between 5.5-8.5, and leavening of the present invention meets professional standard.It should be noted that this Stalk and cow dung superhigh temperature decomposing agent prepared by inventive embodiments 2-5 equally has an above-mentioned test effect, poor between each embodiment It is anisotropic not notable.
The detection of the quality index of 14 stalk of the present invention of embodiment and cow dung superhigh temperature decomposing agent
Using the stalk of the preparation of the embodiment of the present invention 1 and cow dung superhigh temperature decomposing agent as sample, according to《GB/T 19524.1— The measurement of excrement colibacillus group in 2004 fertilizer》、《The measurement of induced worm egg death rate in 19524.2-2004 fertilizer of GB/T》、《NY/ T 1978-2010 fertilizer mercury, arsenic, every, the measurement of lead, chromium content》Etc. national standards detection method to the technical indicator of organic fertilizer It is detected with innoxious index, as a result such as table 3, table 4：
Table 3：The technical indicator of biological organic fertilizer
Project Standard value《NY884-2012》 Measured value
Living bacteria count (cfu/g) ≥0.2 6×1011
Organic matter (in terms of butt, %) ≥40 46.27
Moisture (%) ≤30 20.5
pH 5.5-8.5 7.3
The term of validity (moon) ≥6 12
Table 4：The innoxious index of biological organic fertilizer
Project Standard value《NY884-2012》 Measured value
Excrement colibacillus group number (a/g) ≤100 50
Induced worm egg death rate (%) ≥95 99
Total arsenic (As) (in terms of butt, mg/kg) ≤15 9
Total cadmium (Cd) (in terms of butt, mg/kg) ≤3 2
Total lead (Pb) (mg/kg in terms of butt) ≤50 39
Total chromium (Cr) (in terms of butt, mg/kg) ≤150 99
Total mercury (Hg) (in terms of butt, mg/kg) ≤2 0.8
The above testing result shows the technical indicator of stalk prepared by the present invention and cow dung superhigh temperature decomposing agent and innoxious Index reaches or is better than Ministry of Agriculture's biological organic fertilizer mass calibration, for directly using stalk and cow dung it is quick, efficiently carry out high temperature Compost fermentation opens a new shortcut to prepare high quality biological organic fertilizer, has preferable advanced and practicability.
It should be noted that stalk and cow dung superhigh temperature decomposing agent prepared by 2-5 of the embodiment of the present invention equally have above-mentioned examination Effect is tested, otherness is not notable between each embodiment.
15 stalk of the present invention of embodiment and cow dung the organic fertilizer using effect in rape cultivation are tested
1 test material：
1.1 test plant：Rape
1.2 for trying fertilizer：Stalk and cow dung superhigh temperature organic fertilizer prepared by the embodiment of the present invention 1, commercially available organic fertilizer, chemical fertilizer
2 test methods：
2.1 test process：Experiment is divided into four groups, i.e. T1 is not apply fertilizer；T2 is chemical fertilizer；T3 is commercially available organic fertilizer；T4 is this Stalk and cow dung superhigh temperature decomposing agent prepared by inventive embodiments 1
2.2 test procedure：Respectively by T1, tetra- groups of fertilizer of T2, T3, T4 are according to fertilizer and soil quality percentage 0.05%- 0.15% is added in basin dress soil, and each basin dress soil fills soil 8kg, (the complete phase of composition of the soil that tetra- groups of T1, T2, T3, T4 The rapeseed for choosing full seed together), carries out sowing field planting respectively, and 10 seeds, three weights of every group of soil fertility quality are sowed per basin Multiple parallel laboratory test group.50 days after rape seed, the indexs such as each processing yield, plant height, total nitrogen content and soil fertility, detection are measured As a result as illustrated in tables 5-6.
3 results and analysis：
Table 5 respectively handles rape harvest phase yield and quality
Table 6 respectively handles rape harvest phase soil fertility
As can be seen from Table 5, best, the performance optimal of T4 superhigh temperature stalk and cow dung organic fertilizer processing growth of rape, oil The yield of dish, plant height, chlorophyll content are above chemical fertilizer and the processing of commercially available organic fertilizer.Illustrate stalk of the present invention and cow dung superhigh temperature The composting production of decomposing microbial inoculum fermentation significantly improves soil fertility, and contributes to growth of rape, improves yield of rape.
As can be seen from Table 6, T4 handles the rape harvest phase soil organism, hydrolyzable nitrogen, available phosphorus, quick-acting potassium content difference 7.46%, 31.1%, 36.3%, 42% is increased than not fertilizer treatment；Increase 6.64% than chemical fertilizer processing, 27.7%, 7.7%, 53%；4.71%, 15.4%, 12.6%, 21% is increased than the processing of commercially available organic fertilizer.
Yield of rape is picked and recorded to grouping individually, using the new multipole difference methods of inspection of the Duncan ' s of DPS softwares to data It is analyzed and (is shown in Table 7, table 8)：
Table 7：Analysis of variance table
Become because Quadratic sum Degree of freedom It is square F values Conspicuousness
Between processing 1.4811 3 0.4937 1519.077 Extremely significantly
In processing 0.0026 8 0.0003
Total variation 1.4837 11
Table 8：Yield result duncan's new multiple range method schedule
Note:The significance of difference of the lowercase letter in 5% level, capitalization indicate the difference in 1% level Conspicuousness.
F values (3,8)=1519.077 as shown in Table 7>F0.01 (3,8)=7.59, i.e., difference is extremely notable between different disposal, The effect being significantly improved to yield of rape using stalk and cow dung organic fertilizer.In 1% and 5% level of difference (table 8), processing The yield of rape of 4 (applying stalk and cow dung superhigh temperature decomposed manure) and 2 (to fertilize) of processing are obviously than using commercially available organic fertilizer Processing 3 and the processing 1 do not applied fertilizer have extremely significant raising.Processing 1, processing 2, processing 3 and 4 yield of rape of processing are in 5% water It is gentle extremely notable in 1% horizontal upper difference, wherein applying stalk and cow dung superhigh temperature decomposed manure yield highest.
Chlorophyll in Rape content is picked and recorded to grouping individually, utilizes the new multipole difference methods of inspection of the Duncan ' s of DPS softwares Data are analyzed and (are shown in Table 9, table 10)：
Table 9：Analysis of variance table
Become because Quadratic sum Degree of freedom It is square F values Conspicuousness
Between processing 212.9624 3 70.9875 283.95 Extremely significantly
In processing 2 8 0.25
Total variation 214.9624 11
Table 10：Chlorophyll content result duncan's new multiple range method schedule
Processing Mean value 5% level of signifiance The 1% pole level of signifiance
Processing 4 35.6 a A
Processing 3 33.8 b B
Processing 2 31.2 c C
Processing 1 24.5 d D
Note:The significance of difference of the lowercase letter in 5% level, capitalization indicate the difference in 1% level Conspicuousness.
F values (3,8)=283.95 as shown in Table 9>F0.01 (3,8)=7.59, i.e., difference is extremely notable between different disposal, applies The effect being significantly improved to Chlorophyll in Rape content with stalk and cow dung superhigh temperature organic fertilizer.In 1% and 5% level of difference (table 10), the Chlorophyll in Rape for handling 3 (applying commercially available organic fertilizer) of 4 (applying stalk and cow dung superhigh temperature decomposed manure) and processing contain The processing 1 measured the apparent processing 2 than using chemical fertilizer and do not applied fertilizer has extremely significant raising.Processing 1, processing 2, processing 3 and processing 4 Chlorophyll in Rape content 5% it is horizontal and in 1% horizontal upper difference it is extremely notable, wherein applying stalk and cow dung superhigh temperature is decomposed organic Fertile chlorophyll content highest.
Rape plant height is picked and recorded to grouping individually, using the new multipole difference methods of inspection of the Duncan ' s of DPS softwares to data It is analyzed and (is shown in Table 11, table 12)：
Table 11：Analysis of variance table
Become because Quadratic sum Degree of freedom It is square F values Conspicuousness
Between processing 4130.25 3 1376.75 1694.462 Extremely significantly
In processing 6.5 8 0.8125
Total variation 4136.75 11
Table 12：Plant height result duncan's new multiple range method schedule
Processing Mean value 5% level of signifiance The 1% pole level of signifiance
Processing 4 82 a A
Processing 2 78 b B
Processing 3 60 c C
Processing 1 35 d D
Note:The significance of difference of the lowercase letter in 5% level, capitalization indicate the difference in 1% level Conspicuousness.
F values (3,8)=1694.462 as shown in Table 11>F0.01 (3,8)=7.59, i.e., difference is extremely notable between different disposal, The effect being significantly improved to rape plant height using stalk and cow dung organic fertilizer.In 1% and 5% level of difference (table 12), place It manages 4 (applying stalk and cow dung superhigh temperature decomposed manure) and the rape plant height of 2 (to fertilize) of processing is obviously commercially available more organic than using The processing 3 of fertilizer and the processing 1 that do not apply fertilizer have extremely significant raising.Processing 1, processing 2, processing 3 and 4 yield of rape of processing are 5% It is horizontal and extremely notable in 1% horizontal upper difference, wherein applying stalk and cow dung superhigh temperature decomposed manure rape plant height highest.
Rape accumulation amount of nitrogen sucking is individually picked and is recorded in grouping, utilizes the new multipole difference methods of inspection of the Duncan ' s of DPS softwares Data are analyzed and (are shown in Table 13, table 14)：
Table 13：Analysis of variance table
Table 14：Rape accumulates amount of nitrogen sucking result duncan's new multiple range method schedule
Processing Mean value 5% level of signifiance The 1% pole level of signifiance
Processing 3 16.32 a A
Processing 4 10.86 b B
Processing 2 8.95 b B
Processing 1 3.58 c C
Note:The significance of difference of the lowercase letter in 5% level, capitalization indicate the difference in 1% level Conspicuousness.
F values (3,8)=82.98 as shown in Table 13>F0.01 (3,8)=7.59, i.e., difference is extremely notable between different disposal, applies The effect being significantly improved with commercially available organic fertilizer rape accumulation amount of nitrogen sucking.In 1% and 5% level of difference (table 14), processing 4 (applying stalk and cow dung superhigh temperature decomposed manure) and the rape accumulation amount of nitrogen sucking place that obviously ratio does not apply fertilizer for handling for 2 (to fertilize) Reason 1 has extremely significant raising.It handles 4 (applying stalk and cow dung superhigh temperature decomposed manure) and handles the rape of 2 (to fertilize) and tire out Above difference is not notable in 5% level and in 1% level for product amount of nitrogen sucking.
The random detection soil organic matter content that fetches earth, using the new multipole difference methods of inspection of the Duncan ' s of DPS softwares to data It is analyzed and (is shown in Table 15, table 16)：
Table 15：Analysis of variance table
Become because Quadratic sum Degree of freedom It is square F values Conspicuousness
Between processing 100.4139 3 33.4713 133.885 Significantly
In processing 2 8 0.25
Total variation 102.4139 11
Table 16：Soil organic matter content result duncan's new multiple range method schedule
Processing Mean value 5% level of signifiance The 1% pole level of signifiance
Processing 4 27.02 a A
Processing 3 22.31 b B
Processing 2 20.38 c C
Processing 1 19.56 c C
Note:The significance of difference of the lowercase letter in 5% level, capitalization indicate the difference in 1% level Conspicuousness.
F values (3,8)=133.885 as shown in Table 15>F0.01 (3,8)=7.59, i.e., difference is extremely notable between different disposal, The effect being significantly improved using stalk and the cow dung superhigh temperature Organic Manure on Soil content of organic matter.In 1% and 5% level of difference Upper (table 15) handles 4 (applying stalk and cow dung superhigh temperature decomposed manure) and handles the soil organism of 3 (applying commercially available organic fertilizer) Content obviously than using chemical fertilizer processing 2 and the processing 1 do not applied fertilizer have extremely significant raising.2 soil organism of processing 1 and processing Content 5% it is horizontal and in 1% horizontal upper difference it is not notable, wherein applying stalk and cow dung superhigh temperature decomposed manure soil is organic Matter content highest.
The random detection soil hydrolysis nitrogen content that fetches earth, using the new multipole difference methods of inspection of the Duncan ' s of DPS softwares to data It is analyzed and (is shown in Table 17, table 18)：
Table 17：Analysis of variance table
Become because Quadratic sum Degree of freedom It is square F values Conspicuousness
Between processing 1785.751 3 595.2503 148.813 Significantly
In processing 32 8 4
Total variation 1817.751 11
Table 18：Soil hydrolyzes nitrogen content result duncan's new multiple range method schedule
Processing Mean value 5% level of signifiance The 1% pole level of signifiance
Processing 4 173.6 a A
Processing 3 158.2 b B
Processing 2 145.9 c C
Processing 1 142.5 c C
Note:The significance of difference of the lowercase letter in 5% level, capitalization indicate the difference in 1% level Conspicuousness.
F values (3,8)=148.813 as shown in Table 17>F0.01 (3,8)=7.59, i.e., difference is extremely notable between different disposal, The effect being significantly improved using stalk and cow dung superhigh temperature Organic Manure on Soil hydrolysis nitrogen content.In 1% and 5% level of difference Upper (table 18) handles 4 (applying stalk and cow dung superhigh temperature decomposed manure) and handles the soil hydrolyzable nitrogen of 3 (applying commercially available organic fertilizer) Content obviously than using chemical fertilizer processing 2 and the processing 1 do not applied fertilizer have extremely significant raising.2 soil hydrolyzable nitrogens of processing 1 and processing Above difference is not notable in 5% level and in 1% level for content, wherein applying stalk and the hydrolysis of cow dung superhigh temperature decomposed manure soil Nitrogen content highest.
The random detection soil available phosphorus content that fetches earth, using the new multipole difference methods of inspection of the Duncan ' s of DPS softwares to data It is analyzed and (is shown in Table 19, table 20)：
Table 19：Analysis of variance table
Become because Quadratic sum Degree of freedom It is square F values Conspicuousness
Between processing 2204.55 3 734.85 18371.29 Extremely significantly
In processing 0.32 8 0.04
Total variation 2204.87 11
Table 20：Available phosphorus content result duncan's new multiple range method schedule
Processing Mean value 5% level of signifiance The 1% pole level of signifiance
Processing 4 99.1 a A
Processing 2 91.4 b B
Processing 3 86.5 c C
Processing 1 62.8 d D
Note:The significance of difference of the lowercase letter in 5% level, capitalization indicate the difference in 1% level Conspicuousness.
F values (3,8)=18371.29 as shown in Table 19>F0.01 (3,8)=7.59, i.e., difference is extremely notable between different disposal, The effect being significantly improved using stalk and cow dung Organic Manure on Soil available phosphorus content.(the table in 1% and 5% level of difference 20) 4 (applying stalk and cow dung superhigh temperature decomposed manure), are handled and handle the soil available phosphorus content of 2 (to fertilize) and are obviously compared There is extremely significant raising using the processing 3 and the processing 1 do not applied fertilizer of commercially available organic fertilizer.Processing 1,4 oil of processing 2, processing 3 and processing Above difference is extremely notable in 5% level and in 1% level for dish yield, wherein applying stalk and cow dung superhigh temperature decomposed manure soil has Imitate phosphorus content highest.
The random detection Soil Available potassium content that fetches earth, using the new multipole difference methods of inspection of the Duncan ' s of DPS softwares to data It is analyzed and (is shown in Table 21, table 22)：
Table 21：Analysis of variance table
Become because Quadratic sum Degree of freedom It is square F values Conspicuousness
Between processing 4950 3 1650 66 Significantly
In processing 200 8 25
Total variation 5150 11
Table 22：Soil available nitrogen content results duncan's new multiple range method schedule
Processing Mean value 5% level of signifiance The 1% pole level of signifiance
Processing 4 136 a A
Processing 3 115 b B
Processing 1 94 c C
Processing 2 83 c C
Note:The significance of difference of the lowercase letter in 5% level, capitalization indicate the difference in 1% level Conspicuousness.
F values (3,8)=66 as shown in Table 21>F0.01 (3,8)=7.59, i.e., difference is extremely notable between different disposal, using straw The effect that stalk and cow dung superhigh temperature Organic Manure on Soil quick-acting potassium content are significantly improved.(the table in 1% and 5% level of difference 22) 4 (applying stalk and cow dung superhigh temperature decomposed manure), are handled and handle the Soil Available potassium content of 3 (applying commercially available organic fertilizer) Apparent processing 2 than using chemical fertilizer and the processing 1 that do not apply fertilizer have extremely significant raising.2 Soil Available potassium contents of processing 1 and processing 5% it is horizontal and in 1% horizontal upper difference it is not notable, wherein applying stalk and cow dung superhigh temperature decomposed manure soil available nitrogen contains Measure highest.
Above-mentioned test result shows that the stalk that the present invention obtains and cow dung organic fertilizer replace chemical fertilizer that can significantly improve rape Yield and quality also has remarkable result in terms of improving nitrogen utilization efficiency and active soil organic matter.Certainly, it can also be applied to peppery The cultivation of the vegetables such as green pepper, eggplant, Chinese cabbage equally has said effect, is not illustrating one by one.
It should be noted that stalk and cow dung superhigh temperature decomposing agent prepared by 2-5 of the embodiment of the present invention equally have above-mentioned examination Effect is tested, otherness is not notable between each embodiment.

Claims (10)

1. a kind of stalk and cow dung superhigh temperature decomposing agent are obtained by being dried after stalk, cow dung and decomposed composite bacteria agent mixed fermentation , it is characterised in that：The decomposed composite bacteria agent is made of the raw material of following parts by weight：Thermophilic bacteria agent 1-10 parts, lichens 1-6 parts of gemma bacillus agent, 1-6 parts of aspergillus oryzae microbial inoculum, 1-6 parts of lactobacillus plantarum microbial inoculum, 1-6 parts of thermoactinomyces sacchari microbial inoculum, 1-6 parts of brevibacterium frigoritolerans microbial inoculum.
2. a kind of stalk as described in claim 1 and cow dung superhigh temperature decomposing agent, it is characterised in that：The decomposed composite bacteria agent It is made of following parts by weight raw material：3-8 parts of Thermophilic Bacteria, 2-5 parts of bacillus licheniformis, 2-5 parts of aspergillus oryzae, lactobacillus plantarum 2- 5 parts, 2-5 parts of thermoactinomyces sacchari, 2-5 parts of brevibacterium frigoritolerans.
3. a kind of stalk as claimed in claim 1 or 2 and cow dung superhigh temperature decomposing agent, it is characterised in that：
The preparation method of the thermophilic bacteria agent, includes the following steps：Thermophilic Bacteria is inoculated in the sub- solid slope training of high-temperature strain It supports in base A, after 60-90 DEG C of culture 18-22h, thalline is eluted, obtains elution bacterium solution, the cell concentration for adjusting bacterium solution is 4-6×1011Cfu/mL is inoculated according to inoculum concentration 50-150mL/kg culture mediums in solid medium B, is mixed, 60-90 DEG C stationary culture 16-20h, 1-3 times/day of moisturizing obtain thermophilic bacteria agent, the thermophilic bacteria agent content after drying, crushing ≥6×1011Cfu/g, the Thermophilic Bacteria deposit number are CGMCC No.15721；
The sub- solid medium A groups of high-temperature strain become：Yeast extract 2-3g/L, bacto peptone 2-3g/L, glucose 2-3g/L, PIPES 6-8g/L, plant gel 8-12g/L, agar powder 8-12g/L, surplus are water, adjust pH value 7.2-7.4, 121 DEG C of sterilizing 30min；
The solid medium B groups become：100-120 parts of rice husk, 100-110 parts of wheat bran, 100-120 parts of corn flour, beancake powder 100-115 parts, 100-110 parts of glucose, 2-3 parts of calcium sulfate, 2-4 parts of calcium oxide, 1-3 parts of dipotassium hydrogen phosphate, magnesium sulfate 0.5- 1.2 parts, according to solid material total amount and water weight ratio：5-6:Water, 121 DEG C of sterilizing 45min are added into solid material by 4-5.
4. a kind of stalk as claimed in claim 1 or 2 and cow dung superhigh temperature decomposing agent, it is characterised in that：The lichens gemma The preparation method of bacillus microbial inoculum, includes the following steps：Bacillus licheniformis is inoculated in NA seed solid mediums C, 30-40 DEG C stationary culture 1-2d, thalline is eluted, and obtains elution bacterium solution, and adjustment bacterial concentration is 3-5 × 1010Cfu/mL, according to Inoculum concentration 50-120mL/kg culture mediums, are inoculated in solid medium D, are stirred, 30-40 DEG C of stationary culture 1-2d, through dry After dry, crushing, bacillus licheniformis agent, microbial inoculum content >=4 × 10 are obtained10cfu/g；
The group of the NA seeds solid medium C becomes：Peptone 10-12g/L, powdered beef 3-4g/L, sodium chloride 10-12g/L, Agar 18-20g/L, surplus are water, and adjustment pH value is 7.0-7.2,121 DEG C of sterilizing 30min；
The group of the solid medium D becomes：250-275 parts of rice husk, 500-750 parts of wheat bran, 375-625 parts of corn flour, soya-bean cake 200-275 parts of powder, 225-250 parts of starch, 250-275 parts of laterite, 50-62 parts of glucose, 12-15 parts of calcium sulfate, calcium oxide 15- 20 parts, 5-10 parts of dipotassium hydrogen phosphate, 5-6 parts of magnesium sulfate is 7-15 according to solid material total amount and water weight ratio:2-5 is to solid material Middle addition water mixing, 121 DEG C of sterilizing 45min；
The bacillus licheniformis is bacillus licheniformis CICC 10037, bacillus licheniformis CICC 10084, lichens gemma Any one or a few in bacillus CICC 10085, bacillus licheniformis CICC 10092.
5. a kind of stalk as claimed in claim 1 or 2 and cow dung superhigh temperature decomposing agent, it is characterised in that：
The preparation method of the aspergillus oryzae microbial inoculum, includes the following steps：Aspergillus oryzae strain is inoculated in PDA solid mediums, 25-30 DEG C of stationary culture 5-6d, thalline is eluted, and obtains elution bacterium solution, and adjustment bacterial concentration is 2-4 × 1011Cfu/mL, It according to inoculum concentration 50-120mL/kg culture mediums, is inoculated in solid medium B, 25-30 DEG C of stationary culture 5-6d, through drying, powder After broken, aspergillus oryzae viable bacteria microbial inoculum, microbial inoculum content >=8 × 10 are obtained10cfu/g；
The aspergillus oryzae is aspergillus oryzae CICC 2001, aspergillus oryzae CICC 2011, aspergillus oryzae CICC 2016, aspergillus oryzae CICC Any one or a few in 2022.
6. a kind of stalk as claimed in claim 1 or 2 and cow dung superhigh temperature decomposing agent, it is characterised in that：
The preparation method of the lactobacillus plantarum microbial inoculum, includes the following steps：Lactobacillus plantarum is inoculated in seed solid culture In base C, 30-40 DEG C of stationary culture 2-3d elutes thalline, obtain elution bacterium solution, adjustment bacterial concentration for 4-5 × 1010Cfu/mL is inoculated in by bacterium solution according to inoculum concentration 50-120mL/kg culture mediums in solid medium F, and 30-40 DEG C stands training 2-3d is supported, lactobacillus plantarum microbial inoculum, microbial inoculum content >=8 × 10 are obtained after drying and crushing10Cfu/g, the lactobacillus plantarum tool Body is lactobacillus plantarum CGMCC1.2437；
The preparation method of the solid medium F is：300-500 parts of beancake powder, 300-500 parts of rice husk, 100-200 parts of cotton dregs, 2-5 parts of peptone, 1-5 parts of glucose, 0.1-0.5 parts of magnesium sulfate, according to solid material and water quality ratio 7-10:4-6 is to solid material In plus water, uniformly mix, 121 DEG C sterilizing 45min.
7. a kind of stalk as claimed in claim 1 or 2 and cow dung superhigh temperature decomposing agent, it is characterised in that：
The preparation method of the thermoactinomyces sacchari microbial inoculum, includes the following steps：Thermoactinomyces sacchari is inoculated in NA seeds to consolidate In body culture medium C, 50-70 DEG C of stationary culture 1-2d elutes thalline, obtains elution bacterium solution, and adjustment bacterial concentration is 5- 5.5×1011Cfu/mL is inoculated in solid medium A, 50-70 DEG C by bacterium solution according to inoculum concentration 50-120mL/kg culture mediums Stationary culture 4-5d obtains thermoactinomyces sacchari viable bacteria microbial inoculum, microbial inoculum content >=7 × 10 after drying, crushing11cfu/g；It is described Thermoactinomyces sacchari is specially thermoactinomyces sacchari CGMCC NO.13928.
8. a kind of stalk as claimed in claim 1 or 2 and cow dung superhigh temperature decomposing agent, it is characterised in that：
The preparation method of the brevibacterium frigoritolerans microbial inoculum, includes the following steps：Brevibacterium frigoritolerans are inoculated in the training of NA seed solids Support in base C, 8-10 DEG C of stationary culture 2-3d elutes thalline, obtains elution bacterium solution, adjustment bacterial concentration for 8-9 × 1010Cfu/mL is inoculated in by bacterium solution according to inoculum concentration 50-110mL/kg culture mediums in solid medium G, and 8-10 DEG C stands training It supports 3-4d and obtains brevibacterium frigoritolerans microbial inoculum, microbial inoculum content >=3 × 10 after drying, crushing11cfu/g；The brevibacterium frigoritolerans Deposit number is No.15722；
The group of the solid medium G becomes：Wheat bran 600-620g/L, rice husk 180-200g/L, groundnut meal 80-100g/L, Dregs of beans 100-110g/L, magnesium sulfate 0.5-0.8g/L, ammonium sulfate 3-5g/L, surplus are water, 121 DEG C of sterilizing 45min.
9. the preparation method of stalk and cow dung superhigh temperature decomposing agent as described in claim 1-8 is any, steps are as follows：
(1) microbial inoculum mixes：According to above-mentioned parts by weight by thermophilic bacteria agent, bacillus licheniformis agent, aspergillus oryzae microbial inoculum, plant Lactobacillus microbial inoculum, thermoactinomyces sacchari microbial inoculum, brevibacterium frigoritolerans microbial inoculum, are mixed, and decomposed composite bacteria agent is obtained；
(2) by fresh straw and cow dung according to mass ratio 10-20：1-5 is mixed, and obtains hybrid reactor body, the hybrid reactor body contains Water 55%-65%；
(3) inoculum concentration of the decomposed composite bacteria agent 1-2 ‰ in mass ratio is inoculated into hybrid reactor body, is stirred, carried out Decomposed processing, compost 10-15d obtain stalk and cow dung superhigh temperature decomposing agent.
10. the purposes of stalk and cow dung superhigh temperature decomposing agent in fruits and vegetables field of planting as described in claim 1-8 is any.
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