CN108285882A - A kind of degrading nicotine, decomposition tobacco rod offal, the bacterial strain and method that Chicken dung pig manure is organic fertilizer - Google Patents
A kind of degrading nicotine, decomposition tobacco rod offal, the bacterial strain and method that Chicken dung pig manure is organic fertilizer Download PDFInfo
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Abstract
The invention discloses a kind of degrading nicotine, decomposition tobacco rod offal, the bacterial strains and method that Chicken dung pig manure is organic fertilizer.The bacterial strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and preserving number is CGMCC No.13343, is named as bacillus subtilis Z03 bacterial strains.In terms of nicotine of the present invention in fast degradation tobacco rod offal, Chicken dung pig manure, decomposition tobacco rod wood fibre, there is significant advantage and technological progress.
Description
Technical field
The invention belongs to field of agricultural microbial technology, and in particular to a kind of degrading nicotine, decomposition tobacco rod offal, decomposed chicken
Excrement pig manure is the bacterial strain and method of organic fertilizer.
Background technology
After tobacco seed growing area is gathered in because of tobacco leaf, a large amount of tobacco rods are left in vega, there are about 280-for the annual output of China's tobacco rod
3000000 tons, close to the yield of tobacco leaf.Offal is the waste after Cigarette processing, and China's annual output is up to 600,000 tons, offal heap
Air is directly polluted if there is wind to rise in product such as mountain.Tobacco rod has volume big with offal, and floor space is wide, and value is not high, and nicotine contains
Measure high feature.If the processing method of tobacco rod offal is improper, for example, using burning method, air pollution can be greatly increased, this
It is disagreed with beautiful rural policy route is built.Tobacco rod offal have abundant organic matter, best resource processing method be by
They are degraded to organic fertilizer, realize cycle production.However, the decomposition of tobacco rod offal is faced compared with hang-up.First, tobacco rod quality is tight
Close, wood fibre thickness, crushing difficulty is big, of high cost.Second is that tobacco rod offal nicotine content is high, average content is about 1.5-
2.0%, some flue-cured tobacco cultivars nicotine contents are up to 3.5-4.5%, and because of nicotine content height, organic fertilizer applies crop varieties
Range and land area are severely limited.Nicotine is not only harmful to higher mammal, is also harmful to lower animal, therefore is also agriculture
The main component of industry insecticide.Nicotine insecticide has excellent absorbability and wide, the lasting period length of efficient, low toxicity, insecticidal spectrum etc.
Feature, but inhibiting effect is extended with to the root system of many crops.Third, tobacco rod offal nitrogen is insufficient, the decomposed degradation of single raw material
The requirement of organic fertilizer nitrogen is not achieved.Just because of these problems, there is the risk being incinerated in tobacco rod.High-nicotine organic fertilizer is improper
Using nicotine can cause long-acting adverse effect in field persistent accumulation, to current year plantation and follow-up long-term cropping, gently then subtract
Production, large area is caused when serious, and No kernels or seeds are gathered, as in a year of scarcity, seriously destroys soil, influences soil sustainable development.
About the research of tobacco rod offal, it is still within the starting stage at present, predominantly stays in the sieve of single nicotine degradation bacterium
Optimization of the choosing with condition of culture or the research in terms of single tobacco rod decomposition.Commercially available straw decomposing leavening, lacks to cigarette
Alkaline degradation systematic research lacks the effective ways of comprehensive treatment tobacco rod offal.Some leavening energy degrading straw fibers, but cigarette
Alkali tolerance is inadequate, influences growth breeding and is even suppressed.Some nicotine degradation bacterium are not resistant to 60-80 DEG C of hot fermentations,
It is less desirable to the harmless treatment of the decomposition effect, animal waste of stalk.
Invention content
The present invention is directed to overcome the deficiencies in the prior art, a kind of degrading nicotine of this offer, decomposition tobacco rod offal, Chicken dung
Pig manure is the bacterial strain and method of organic fertilizer.This method can fully decomposition tobacco rod offal, Chicken dung pig manure, degrading nicotine, obtain
Fertilizer efficiency is up to standard, nicotine content is low, and the organic fertilizer non-hazardous to soil, crop, realizes flue-cured tobacco waste, animal waste
Etc. the purpose to turn waste into wealth.Realize farming and animal husbandry pollution sources recycling, innoxious use.
In order to achieve the above object, technical solution provided by the invention is：
The degrading nicotine, decomposition tobacco rod offal, the bacterial strain that Chicken dung pig manure is organic fertilizer are preserved in China Microbiological
Culture presevation administration committee common micro-organisms center, preserving number are CGMCC No.13343, are named as bacillus subtilis
Z03 bacterial strains.
Wherein, the resistance to nicotine of the bacterial strain, degrading nicotine, decomposition tobacco rod offal, Chicken dung pig manure.The bacterial strain is suitble to give birth to
Long temperature is 37 DEG C -45 DEG C, and tolerable temperature is 65 DEG C -78 DEG C.The bacterial strain is 3.0-in nicotine mass percent concentration
Energy normal growth breeding in 4.0% culture medium, per mL zymotic fluid viable counts up to 20,000,000,000 or more；In nicotine mass percent
It can preferably be grown in a concentration of 6.0-8.0% culture medium, per mL zymotic fluid viable counts up to 10-40 hundred million；In nicotine quality
In the culture medium of 10.0% or more percent concentration, growth is suppressed, per mL zymotic fluid viable counts less than 1.0 hundred million.It is described
Bacterial strain is in the fluid nutrient medium that nicotine mass percent concentration is 3.0-4.0%, and shaking speed 200r/m, 37 DEG C ferment 4 days
Nicotine residual quantity is less than 0.1%；In the solid medium that nicotine mass percent concentration is 2.0-2.5%, 37 DEG C of fermentations 3
Its nicotine residual quantity is less than 0.09%.
Included the following steps using the method that above-mentioned bacterial strains prepare liquid fermentation agent：
(1) preparation of shake-flask seed：The strain tube inclined-plane is inoculated with the how bottled container for having seed culture medium respectively
In, seed culture based formulas is：Yeast extract 0.25-0.35%, preferably 0.3%, soyabean expeller 0.25-0.35%, preferably
0.3%, glucose 1.5-2.5%, preferably 2%, pH 7.0,200-300mL of water, preferably 250mL；In 37 DEG C, 200r/
Ferment 13-15h under the conditions of m, obtains seed liquor, and bacterium number is up to 100-200 hundred million/mL；Percentage in the fermentative medium formula
Than being the component shared mass percent in the fermentation medium；
(2) preparation of secondary liquid seed：Seed liquor is seeded in the seed culture medium of fermentation tank, the seed liquor with
The volume ratio of fermentation tank is (2-3):100, preferably 2.5:100, it is planted described in the same step of seed culture medium (1) in fermentation tank
Sub- culture medium, in 37 DEG C, 100r/m ferments 10-12h, obtains secondary seed solution, and bacterium number is up to 100-150 hundred million/mL；.
(3) preparation of liquid starter：Secondary seed solution is seeded in the fermentation medium of fermentation tank, secondary seed solution
Volume ratio with fermentation tank is 1:20, fermentative medium formula is：Yeast extract 0.25-0.35%, preferably 0.3%, soyabean expeller
0.25-0.35%, preferably 0.3%, glucose 1.5-2.5%, GP330 antifoaming agent 0.02-0.04%, pH 7.0, water
1500L；In volume ratio be 1:1 fermentation ventilatory capacity, 40 DEG C, ferment for 24 hours to get liquid fermentation agent, liquid under the conditions of 60r/m
Viable count is 200-300 hundred million/mL in fermenting agent；Percentage in the fermentative medium formula is that the component is being sent out
Shared mass percent in ferment culture medium.
Included the following steps using the method that above-mentioned bacterial strains prepare solid fermentation microbial inoculum：
(1) preparation of shake-flask seed：The strain tube inclined-plane is inoculated with the how bottled container for having seed culture medium respectively
In, seed culture based formulas is：Yeast extract 0.25-0.35%, preferably 0.3%, soyabean expeller 0.25-0.35%, preferably
0.3%, glucose 1.5-2.5%, pH 7.0,200-300mL of water, preferably 250mL；In 37 DEG C, 200r/m conditions issue
13-15h of ferment, obtains seed liquor, and bacterium number is up to 100-200 hundred million/mL；Percentage in the fermentative medium formula is the group
Divide mass percent shared in the fermentation medium；
(2) preparation of two level solid seed：Seed liquor is seeded in solid medium, the seed liquor and solid culture
The mass ratio of base is 1:(45-55), preferably 1:50, the solid culture based formulas is：Wheat bran 40%, soyabean expeller 10%, water
50-60%, pH are natural；In 37 DEG C ferment 36-48h, pneumatic conveying drying to water content be 6-8%, bacterium number be more than 15,000,000,000/
G is to get solid fermentation microbial inoculum.
Above-mentioned bacterial strains can be used for preparing organic fertilizer or soil conditioner.
The degrading nicotine, decomposition tobacco rod offal, the method that Chicken dung pig manure is organic fertilizer are by the cigarette of preliminary crushing
Bar, offal, chicken manure, pig manure mixing, and it is 50-60% to adjust water content, then sprays aforesaid liquid fermenting agent, it is mixed per 1t
It closes object and sprays 1-2kg liquid fermentation agents；Or the above-mentioned solid fermentation microbial inoculum of inoculation, it is inoculated with 1-2kg solids per 1t mixtures
Fermenting agent；Compost treatment is carried out, control compost fermentation temperature, fermentation time, turning number make compost water content be less than
30%, obtain color and luster nigrescence, odorless organic fertilizer；In organic fertilizer：The content of organic matter >=45%, total nutrient >=5%, water content≤
30%, pH5.5-8.5, the worm's ovum death rate >=98%, colibacillus of excrement group number≤50/g；In every kg dry matters, arsenic As≤
30, mercury Hg≤30, lead Pb≤100, cadmium Cd≤3, chromium Cr≤300；Nicotine content is less than 0.1%.
The invention will be further described below：
The present invention provides the bacterium of a kind of resistance to nicotine, degrading nicotine, farming and animal husbandry straw decomposition, excrement high temperature Rapid Fermentation
Strain.
Garden tobacco planting garden is weeded from Agricultural University Of Hunan, is detached in the tobacco rod offal of artificial high temperature pile fermentation, chicken manure pig manure heap
Acquired more than 120 plants of single bacterium colonies, then obtained through hot fermentation secondary screening, high-nicotine degradation secondary screening, bacterial strain Z03 through Morphological Identification and
Molecule sequencing is accredited as bacillus subtilis.Bacillus subtilis Z03 is preserved in China Microbiological on November 25th, 2016
Culture presevation administration committee common micro-organisms center, preserving number：CGMCC No.13343.
In the present invention, the tobacco rod of preliminary crushing, offal, chicken manure, pig manure are mixed by different ratio, adjusts water content to 50
- 60%, while spraying inoculation bacillus subtilis Z03 leavenings (liquid or solid leavening, per kg leavening viable bacterias
Number is more than 10,000,000,000), inoculum concentration is that 1t mixed materials are inoculated with Z03 liquid starters 1.5kg；Or combined inoculation withered grass gemma
Bacillus Z03 solid fermentation agents, inoculum concentration are that 1t mixed materials are inoculated with Z03 solid fermentation agent 1.5kg, and microbial inoculum Z03 is mixed with material
It closes uniformly, organic fertilizer fermentation base-material is made, base-material heap is set to ridge type, mainly using few in heat-resistant bacillus Z03 and base-material
Measure original microorganism raw material fermentation production organic fertilizer.The base-material ridge heap that ferments is 2.0-2.5 meters wide, 0.6-0.8 meter high, length
More than 15 meters.2-3 days before compost, covering with ground sheeting is fermented base-material, and heat and moisture preserving, accelerated warming waits for pile core temperature up to 43 DEG C or more
When, remove mulch；Start turning (forklift or artificial turning), turning in 3 days 1 time, heap temperature when pile core temperature is up to 52 DEG C or more
60 DEG C or more maintain 8 days or more.Summer only needs 14-17 days quick compostings for completing fermentation base-material；Winter only needs 20-25 days
The quick composting of fermentation base-material is completed, nicotine substantially reduces, while the abundant decomposition of nicotine offal, chicken manure pig manure are fully decomposed, hair
The ferment period greatly shortens than Natural compost fermentation process.Compost quality control is as follows：Control compost fermentation temperature, fermentation time,
Turning number makes compost water content be less than 30%, and color and luster blacks, odorless；Its essential characteristic, which is removed, includes organic fertilizer sense organ and physics and chemistry
Index meets outside organic fertilizer new standard NY525-2011, that is, the content of organic matter >=45%, total nutrient >=5%, water content
≤ 30%, pH5.5~8.5, the worm's ovum death rate >=98%, colibacillus of excrement group number≤50/g；In per gram of dry matter, arsenic As
≤ 30, mercury Hg≤30, lead Pb≤100, cadmium Cd≤3, chromium Cr≤300；The present invention also specifically limits nicotine in organic fertilizer and contains
Amount, nicotine content per kilogram are less than 0.1%.
Compared with prior art, nicotine, Chicken dung pig manure, decomposition tobacco rod of the present invention in fast degradation tobacco rod offal
In terms of wood fibre, there is significant advantage and technological progress：
First, efficiently degrading nicotine, leavening component is simplified, ferment effect is quick, is convenient for practical operation.11 table 1 of embodiment
Experimental result can be seen that the present invention Z03 be single fermentation agent, fermentation total time it is most short (22 days), nicotine content is minimum
(being less than 0.1%), reduce time cost.Other leavenings, because the nicotine content of tobacco rod offal is excessively high, it is suppressed that leavening
Growth and breeding to extend fermentation time (extending 5-7 days than expected), thus increases production time cost.
Second is that fermented product not only up to organic fertilizer requirement, and the resource of particularly suitable flue-cured tobacco tobacco rod, offal the like waste
Change, innoxious use, hence it is evident that be better than commercially available stover ferment agent.The experimental result of 12 table 2 of embodiment can be seen that production of the present invention
Organic fertilizer product be not only applicable in vega, but also nicotine fully degraded can also use germination, the plant development of other crops,
The conditioner product use prepared is improving soil texture, and prevention soil block degeneration, enhancing soil water-retaining, ventilation promote
Significant effect in terms of soil microbial activity.
Specific implementation mode
Embodiment 1, the acquisition of bacillus subtilis Z03
From several compost of tobacco rod, offal, chicken manure, pig manure that the artificial high temperature pile fermentation in garden flue-cured tobacco base is weeded by Agricultural University Of Hunan
Separating obtained high temperature resistant, resistance to nicotine bacillus in sampling obtain more than 120 plants of single bacterium colony, wherein bacterial strain Z03, through Morphological Identification
It is sequenced and identifies with molecule, bacterial strain Z03 is bacillus subtilis (Bacillus subtilis).Bacillus subtilis Z03 in
On November 25th, 2016 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number： CGMCC
No.13343。
Embodiment 2, bacillus subtilis Z03 growths and production gemma characteristic measurement
It is several (250mL triangular flasks) to prepare fluid nutrient medium 100mL, culture medium prescription is yeast extract 0.5%, glucose
Sterilize 25min under the conditions of 7.0,121 DEG C of 2.0%, pH.By the test tube slants bacterial strain Z03, hair not Jie Zhong in fluid nutrient medium,
30 DEG C, 35 DEG C, 37 DEG C, 39 DEG C, 42 DEG C, 45 DEG C, 48 DEG C, 52 DEG C, 55 DEG C, 60 DEG C, 65 DEG C, 70 DEG C, 75 DEG C of shaking tables or concussion water
It is cultivated in bath, measures viable count and gemma number.Z03 bacterial strains are suitble to growth temperature to be 37 DEG C -45 DEG C, and every milliliter of viable count reaches
20000000000 or more, every milliliter up to 2,000,000,000 or more of gemma number；Tolerable temperature is 65 DEG C -78 DEG C.
Embodiment 3, the preparation of bacillus subtilis Z03 liquid shaking bottle seeds
The preparation of shake-flask seed, by the test tube slants bacterial strain Z03 every time one ring of inoculation respectively in 10 bottles of 500mL triangular flasks,
Culture medium prescription is yeast extract 0.3%, soyabean expeller 0.3%, glucose 2.0%, pH 7.0, tap water 250mL, 121 DEG C of sterilizings
25min；37 DEG C, 200r/m ferments 13-15 hours, and shake-flask seed is made up to 100-200 hundred million/mL in bacterium number.
Embodiment 4, the preparation of bacillus subtilis Z03 secondary seeds
10 bottles of total 2500mL of bacterial strain Z03 shaking flask grades seed are seeded to 100L fermentation tanks by the preparation of secondary seed, culture
Based formulas is with shake-flask seed formula, and 37 DEG C, 100r/m ferments 10-12 hours, and bacterium number is made up to 100-150 hundred million/mL
Secondary seed.
Embodiment 5, the preparation of bacillus subtilis Z03 production seeds
The preparation of bacillus subtilis Z03 (production seed) leavening, by secondary seed by volume 1:The nothing of 20 (v/v)
Bacterium operates in pipeline culture transferring to 2000L fermentation tanks, and culture medium prescription is yeast extract 0.3%, soyabean expeller 0.3%, glucose
2.0%, GP330 antifoaming agent 0.03%, pH 7.0, tap water 1500L, 121 DEG C of sterilizing 30min；Ventilatory capacity (1:1, v/v),
40 DEG C, 60r/m ferments 24 hours, and viable count is 200-300 hundred million/mL, every milliliter up to 20-50 hundred million or more of gemma number.
It can be used as tobacco rod, offal, chicken manure, pig manure fermented liquid fermenting agent.
Embodiment 6, the preparation of bacillus subtilis Z03 solid fermentation microbial inoculums
The preparation of two level solid seed, by bacterial strain Z03 shake-flask seeds, in mass ratio 1:50 (w/w) sterile workings are seeded to
In 200 disk 40cm × 60cm × 5cm stainless steel trays, every disk culture medium 4kg (wet feed)；Culture medium prescription is wheat bran 40%, yellow
Dregs of beans 10%, water 50%, pH is naturally, 124 DEG C of sterilizing 40min；It 37 DEG C, ferments 36-48 hours, pneumatic conveying drying is to aqueous
Amount 6% -8%, viable count are more than 15,000,000,000/g (dry weight), you can as tobacco rod offal-chicken manure pig manure solid fermentation bacterium
Agent.
Embodiment 7, the heat-resistance test of bacillus subtilis Z03 solid fermentation microbial inoculums
By the bacillus subtilis Z03 solid fermentation microbial inoculum 100g that viable count is 15,000,000,000/g, at 80 DEG C of insulating box
Reason 24 hours measures viable count before and after the processing.The result shows that Z03 solid fermentations microbial inoculum is handled 24 hours in 80 DEG C of insulating box
Afterwards, viable count still reaches to 115-120 hundred million/g, and survival rate illustrates fermenting agent high temperature resistant up to 75% or more, is conducive to carry out
During High-Temperature Composting is fermented.
Embodiment 8, the resistance to nicotine performance tests of bacillus subtilis Z03
It is several to prepare 100mL culture mediums, culture medium prescription is yeast extract 1.0%, glucose 4.0%, nicotine 0-10.0%
It differs, pH is naturally, 121 sterilizing 20min, and inoculation bacillus subtilis Z03,37 DEG C, 200r/m ferments 24 hours, measures viable bacteria
Number.When nicotinic density is 0, every milliliter up to 200-300 hundred million of viable count；When nicotinic density 3.0% -4.0%, every milliliter of fermentation
Liquid viable count can be up to 150-200 hundred million or more, and growth is barely affected；The culture medium of nicotinic density 6.0% -8.0%
In can also preferably grow, every milliliter of zymotic fluid viable count can be up to 10-40 hundred million；The culture medium of 10.0% or more nicotinic density
In, strain growth is suppressed, and less than 1.0 hundred million, growth is suppressed every milliliter of zymotic fluid viable count；Illustrate that bacterial strain Z03 is resistance to
Nicotine performance is fine.
Embodiment 9, bacillus subtilis Z03 compost tobacco rod, offal, chicken manure carry out organic fertilizer production experiment
By dry biomass ratio, tobacco rod, offal, chicken manure ratio be 1:1:1 formula hybrid composting material, water content
50% -60%, compost material mixture per ton is inoculated with 1.5 kilograms of bacillus subtilis Z03 solid fermentation agents, and fermenting agent is lived
Bacterium number is more than 15,000,000,000/g；Then base-material ridge formula is stacked again, fermentation base-material is 0.8 meter high per 2.5 meters of row spacing, 20 meters long, base
Material heap covering with ground sheeting starts compost fermentation.Period is more than 1 meter of inserted-link type thermometer with inserted link length and measures pile core temperature, often
It is measured once, and 2 days heap temperature of fermentation take down mulch, forklift turning is used in combination up to 43 DEG C；4 days heap temperature of fermenting are up to 52 DEG C；It holds later
Continuous to be warming up to 60 DEG C or more, up to 70 DEG C, the duration, moisture constantly volatilized during fermentation and turning up to 8 days or more.
Temperature is begun to decline after fermentation 15 days, and by multiple turning, at 20 days, moisture is less than 30%, and fermentation base-material blacks, and foul smell is notable
It reduces, without apparent flue-cured tobacco taste, fermentation is basically completed, and reaches organic fertilizer standard.When fermentation ends, per kilogram organic fertilizer nicotine content is
0.85 gram, 16 grams of the content of per kilogram material nicotine well below fermentation initial stage.
Embodiment 10, bacillus subtilis Z03 compost tobacco rod, offal, mushroom residue, swine excrement carry out organic fertilizer production experiment
By dry biomass ratio, tobacco rod, offal, mushroom residue, pig manure ratio be 1:1:1:1 formula hybrid composting material, water content
50% -60%, compost material mixture per ton is inoculated with 1.5 kilograms of bacillus subtilis Z03 liquid starters, and fermenting agent is lived
Bacterium number is more than 20,000,000,000/g；Then base-material ridge formula is stacked again, fermentation base-material is 0.9 meter high per 2.5 meters of row spacing, 25 meters long, base
Material heap covering with ground sheeting starts compost fermentation.Period is more than 1 meter of inserted-link type thermometer with inserted link length and measures pile core temperature, often
It is measured once, and 2 days heap temperature of fermentation take down mulch, forklift turning is used in combination up to 45 DEG C；4 days heap temperature of fermenting are up to 55 DEG C；It holds later
Continuous to be warming up to 60 DEG C or more, up to 68 DEG C, the duration, moisture constantly volatilized during fermentation and turning up to 8 days or more.
Temperature is begun to decline after fermentation 14 days, and by multiple turning, at 20 days, moisture is less than 30%, and fermentation base-material blacks, and foul smell is notable
It reduces, fermentation is basically completed, and reaches organic fertilizer standard.When fermentation ends, per kilogram organic fertilizer nicotine content is 0.67 gram, per kilogram
12 grams of the content of material nicotine well below fermentation initial stage.
Embodiment 11, bacillus subtilis Z03 test (being shown in Table 1) with commercially available compost fermentation agent comparison production
By dry biomass ratio, tobacco rod, offal, chicken manure ratio be 1:1:1 formula hybrid composting material, water content
50% -60% several ridges of compost, different fermentations agent is inoculated with per ridge respectively, and compost material mixture inoculum concentration per ton is 1.5
Kilogram.Base-material ferment per 2.5 meters of row spacing, 0.8 meter high, 2.5 meters long, base-material heap covering with ground sheeting starts compost fermentation.Period,
It is more than 1 meter of inserted-link type thermometer with inserted link length and measures pile core temperature, measure daily once, records Fermentation Engineering data, fermentation
At the end of, detect nicotine content in organic fertilizer product.It the results are shown in Table 1.As it can be seen from table 1 in addition to control group, experimental group is in reality
It tests fermentation tobacco rod, offal, Chicken Manure Compost in the period and reaches organic fertilizer standard.The palliating degradation degree of nicotine, with bacillus subtilis Z03
It is optimal, degradation rate 94.4%；Secondly, the nicotine degradation rate of spontaneous fermentation is only 48.8% to commercially available leavening B.From fermentation high temperature
From the point of view of duration and abundant decomposed time, optimal with bacillus subtilis Z03, other fermenting agents are due to nicotine shadow in compost
Growth breeding is rung, than estimated longer fermentation times 5-8 days.It can be seen that bacillus subtilis Z03 is suitble to tobacco rod offal special
Leavening.
The influence that 1. different fermentations agent of table ferments to tobacco rod, offal, Chicken Manure Compost
Embodiment 12, influence (be shown in Table 2) of the windrow to Chinese cabbage germination index in composting process
It is control with distilled water, handles offal 1.0% and bacterial strain Z03 the material (cigarette of compost different time points respectively
Bar, offal, chicken manure dry biomass are than 1:1:1) tracing detection of progress Shandong Chinese cabbage seeds germination index, GI (%)=
(percentage of seedgermination × seed of compost extraction is with length)/(percentage of seedgermination × seed of distilled water is with length) × 100, as a result see
Table 2.Untreated offal significantly inhibits effect to Chinese cabbage germination percentage, influences root growth.Compost fermentation initial stage, compost are rotten
It is ripe be it is abundant, nicotine degradation is not thorough, and seriously affects germination.As it can be seen that having using tobacco rod offal as organic matter fermenting and producing
The height of machine fertilizer, nicotine content is the important indicator for evaluating the organic fertilizer, although not having in organic fertilizer standard NY525-2011
This index is limited, but in produce reality application, it is necessary to take in, otherwise can influence crop yield, destroy soil physical chemistry
Characteristic.
The influence of 2. compost material of table and its nicotine to Chinese cabbage germination index
| Germination index (GI)a | 5 days | 10 days | 15 days | 20 days |
| Distilled water (control) | 100% | 100% | 100% | 100% |
| Offal 1.0% | 18.1% | 18.5% | 20.6% | 23.8% |
| Compost material 1.0% | 21.2% | 75.6% | 94.4% | 98.5% |
aNote：If GI>50%, then it is believed that it is substantially decomposed, when GI reaches 80% -85%, this compost
Think completely decomposed, there is no toxicity to plant.
Claims (9)
1. a kind of degrading nicotine, decomposition tobacco rod offal, Chicken dung pig manure are the bacterial strain of organic fertilizer, which is characterized in that the bacterium
Strain is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and preserving number is CGMCC No.13343, life
Entitled bacillus subtilis Z03 bacterial strains.
2. bacterial strain as described in claim 1, which is characterized in that the resistance to nicotine of the bacterial strain, degrading nicotine, decomposition tobacco rod offal,
Chicken dung pig manure.
3. bacterial strain as described in claim 1, which is characterized in that the bacterial strain is suitble to growth temperature to be 37 DEG C -45 DEG C, tolerance
Temperature is 65 DEG C -78 DEG C.
4. bacterial strain as described in claim 1, which is characterized in that the bacterial strain is 3.0-in nicotine mass percent concentration
Energy normal growth breeding in 4.0% culture medium, per mL zymotic fluid viable counts up to 20,000,000,000 or more；In nicotine mass percent
It can preferably be grown in a concentration of 6.0-8.0% culture medium, per mL zymotic fluid viable counts up to 10-40 hundred million；In nicotine quality
In the culture medium of 10.0% or more percent concentration, growth is suppressed, per mL zymotic fluid viable counts less than 1.0 hundred million.
5. bacterial strain as described in claim 1, which is characterized in that the bacterial strain is 3.0-in nicotine mass percent concentration
In 4.0% fluid nutrient medium, shaking speed 200r/m, 37 DEG C of fermentations, 4 days nicotine residual quantities are less than 0.1%；In nicotine quality
In the solid medium that percent concentration is 2.0-2.5%, 37 DEG C of fermentations, 3 days nicotine residual quantities are less than 0.09%.
6. the method for preparing liquid fermentation agent using the bacterial strain as described in any one of claim 1 to 5, which is characterized in that described
Method includes the following steps：
(1) preparation of shake-flask seed：The strain tube inclined-plane is inoculated with respectively in the how bottled container for having seed culture medium, is planted
Sub- culture medium prescription is：Yeast extract 0.25-0.35%, soyabean expeller 0.25-0.35%, glucose 1.5-2.5%, pH 7.0,
200-300mL of water；In 37 DEG C, ferment 13-15h under the conditions of 200r/m, obtains seed liquor, and bacterium number is up to 100-200 hundred million/mL；Institute
It is the component shared mass percent in the fermentation medium to state the percentage in fermentative medium formula；
(2) preparation of secondary liquid seed：Seed liquor is seeded in the seed culture medium of fermentation tank, the seed liquor and fermentation
The volume ratio of tank is (2-3):100, seed culture medium described in the same step of seed culture medium (1) in fermentation tank, in 37 DEG C,
100r/m ferments 10-12h, obtains secondary seed solution, and bacterium number is up to 100-150 hundred million/mL；.
(3) preparation of liquid starter：Secondary seed solution is seeded in the fermentation medium of fermentation tank, secondary seed solution and hair
The volume ratio of fermentation tank is 1:20, fermentative medium formula is：Yeast extract 0.25-35%, soyabean expeller 0.25-35%, glucose
1.5-2.5%, GP330 antifoaming agent 0.02-0.04%, pH 7.0, water 1500L；In volume ratio be 1:1 fermentation ventilatory capacity,
40 DEG C, ferment under the conditions of 60r/m for 24 hours to get liquid fermentation agent, in liquid fermentation agent viable count be 200-300 hundred million/
mL；Percentage in the fermentative medium formula is the component shared mass percent in the fermentation medium..
7. the method for preparing solid fermentation microbial inoculum using the bacterial strain as described in any one of claim 1 to 5, which is characterized in that described
Method includes the following steps：
(1) preparation of shake-flask seed：The strain tube inclined-plane is inoculated with respectively in the how bottled container for having seed culture medium, is planted
Sub- culture medium prescription is：Yeast extract 0.25-0.35%, soyabean expeller 0.25-0.35%, glucose 1.5-2.5%, pH 7.0,
200-300mL of water；In 37 DEG C, ferment 13-15h under the conditions of 200r/m, obtains seed liquor, and bacterium number is up to 100-200 hundred million/mL；Institute
It is the component shared mass percent in the fermentation medium to state the percentage in fermentative medium formula；
(2) preparation of two level solid seed：Seed liquor is seeded in solid medium, the seed liquor and solid medium
Mass ratio is 1:(45-55), the solid culture based formulas are：Wheat bran 40%, soyabean expeller 10%, water 50-60%, pH is certainly
So；Ferment 36-48h in 37 DEG C, and pneumatic conveying drying to water content is 6-8%, and bacterium number is more than 15,000,000,000/g to get solid fermentation
Microbial inoculum.
8. bacterial strain is applied in preparing organic fertilizer or soil conditioner as described in any one of claim 1 to 5.
9. a kind of degrading nicotine, decomposition tobacco rod offal, Chicken dung pig manure are the method for organic fertilizer, which is characterized in that the side
Method is to mix the tobacco rod of preliminary crushing, offal, chicken manure, pig manure, and it is 50-60% to adjust water content, then sprays right and wants
The liquid fermentation agent described in 6 is sought, sprays 1-2kg liquid fermentation agents per 1t mixtures；Or described in inoculation claim 7
Solid fermentation microbial inoculum, 1-2kg solid fermentation microbial inoculums are inoculated with per 1t mixtures；Compost treatment is carried out, compost fermentation temperature is controlled
Degree, fermentation time, turning number make compost water content be less than 30%, obtain color and luster nigrescence, odorless organic fertilizer；In organic fertilizer：Have
Machine matter content >=45%, total nutrient >=5%, water content≤30%, pH5.5-8.5, the worm's ovum death rate >=98%, excrement large intestine bar
Flora number≤50/g；In every kg dry matters, arsenic As≤30, mercury Hg≤30, lead Pb≤100, cadmium Cd≤3, chromium Cr≤300；Cigarette
Alkali content is less than 0.1%.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103351187A (en) * | 2013-07-25 | 2013-10-16 | 梅县福田肥业有限公司 | Method for manufacturing fertilizer and pesticide double-effect organic fertilizer with waste tobacco materials |
| CN106754541A (en) * | 2017-01-09 | 2017-05-31 | 杨丁元 | A kind of tobacco waste fermenting agent and its application method and application |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103351187A (en) * | 2013-07-25 | 2013-10-16 | 梅县福田肥业有限公司 | Method for manufacturing fertilizer and pesticide double-effect organic fertilizer with waste tobacco materials |
| CN106754541A (en) * | 2017-01-09 | 2017-05-31 | 杨丁元 | A kind of tobacco waste fermenting agent and its application method and application |
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