CN102732465A - Thermophilic facultative aerobic bacteria UTM601 and application thereof - Google Patents
Thermophilic facultative aerobic bacteria UTM601 and application thereof Download PDFInfo
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Classifications
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
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Abstract
The invention provides a thermophilic facultative aerobic bacteria (anoxybacillus pushchinoensis) UTM601, whose preservation number is CGMCC No.5929. The bacterial strain UTM601 grows well under micro-aerobic and aerobic environments at 50-70 DEG C, and has the characteristics of being capable of well adapting to compost high-temperature environment with high temperature and inhomogenous oxygen content distribution and degrading organic substances quickly. The invention also provides the method of using the bacterial strain to prepare compost inoculum. The bacterial strain of the invention uses organic waste as carriers to prepare solid-state microorganism compost inoculum, which is low in cost, and the bacterial strain will not influence material moisture when the bacterial strain is applied to compost. With lower multiple horizontal ventilation volume, the solid-state microorganism compost inoculum adopted by the invention for high-temperature compost can realize the high-temperature fermentation of materials and avoid complex ventilation management and control problems during the composting process, and the method has the advantages of simple operation and energy saving.
Description
Technical field
The present invention relates to the environmental microorganism technical field, particularly, relate to a kind of thermophilic double oxygen bacteria strain UTM601 and the application in During High-Temperature Composting thereof.
Background technology
Composting is under microbial process, to make organism mineralization, humify and the innoxious process that becomes the fertilizer that becomes thoroughly decomposed through thermophilic fermentation.During High-Temperature Composting depends on the microbial source of tool some amount and vigor as a kind of microbiological process.Along with industrialization, the commercialization of compost, microorganism inoculum has become an important techniques and has been employed.
At present, the zymotechnique that During High-Temperature Composting adopts is generally aerobic fermentation, and microbial inoculant also mainly is middle temperature aerobic bacteria.Aerobic fermentation must be kept certain oxygen concn.But compost is a complex system, and for the different sites of different batch mixing ratios, heap body, the incidence of air blast process and degree all can change thereupon; Even if under the identical ventilation, the oxygen level distribution of piling different sites in the body is also inconsistent, and this just makes and in actually operating, is difficult to hold suitable ventilation: ventilation is too small; Mikrobe can't obtain enough oxygen; The active reduction, even can be in anaerobic state, the compost reaction process is slow; Ventilation is excessive, and the temperature of heap body is not easy accumulation, is difficult to guarantee innoxious effect, also influences the normal procedure of compost, and increases energy consumption, and nitrogen loses in a large number, reduces the fertilizer benefit of tunning.Especially in 50 ℃-70 ℃ compost pliotherm period environment, the activity of microbial inoculant commonly used also receives the inhibition of temperature.
Thermophilic double oxygen bacterium has thermophilic, amphimicrobian; Than mesophilic bacteria, have higher organic matter degradation active with the advantage of metabolic rate faster, can improve the degradation rate of organic substance; Shorten technology working time, utilize at recycling organic waste to have the potential using value in the field.Thermophilic double oxygen bacterium is applied to compost as Inoculant, will effectively overcomes the above problems, promote the compost process.But also do not see the open report that is applied to During High-Temperature Composting about thermophilic double oxygen bacterium as Inoculant at present.
Summary of the invention
The object of the present invention is to provide a kind of thermophilic double oxygen bacterium (Anoxybacillus pushchinoensis) and application thereof.
Bacterial strain UTM601 of the present invention is from the pliotherm period sludge composting sample that picks up from the green source of the earth environmental protection science and technology (Beijing) ltd compost fermentation workshop; Under 40 ℃; Through little oxygen passage and attenuation; The Anoxybacillus sp. bacterial strain that obtains after separation and purification, the test of aerobism growth performance, the degradation property comparison test screening, called after UTM601.This bacterial strain is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on March 22nd, 2012 and (is called for short CGMCC; Address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City; Institute of Microorganism, Academia Sinica; Postcode 100101), classification called after Anoxybacillus pushchinoensis, preserving number CGMCC No.5929.
Bacterial strain UTM601 of the present invention has following characteristic:
UTM601 is at NA substratum (Carnis Bovis seu Bubali cream 3-5g, peptone 6-10g, sodium-chlor 3-5g, agar 14-18g; Plant gel 7-9g, zero(ppm) water 1000ml, pH7.2) upward colony characteristics is: form is irregular; Surface wettability, light yellow, erode shape; Flat, growth is cultivated the following 36 hours about 3mm of colony diameter for 50-70 ℃ rapidly.Utilize the primer primer to be 27f:5 '-AGA GTT
TGA TCC TGG CTC AG-3 ' and 1492r:5 '-GGT TAC CTT GTT ACG ACTT-3 ' is through the 16S of this bacterial strain of pcr amplification rRNA gene order; The acquisition sequence is submitted to EzTaxon DB (http://www.eztaxon.org/) and carries out the homology comparison, and the result shows and Anoxybacillus pushchinoensis K-1
TSimilarity is the highest, is 98.80%, and this sequence is to identify the principal character foundation of this bacterial strain.Next is above-mentioned bacterial strains well-grown and the accidental quality of the characteristic with quick degraded starch for this bacterial strain of discriminating under little oxygen condition.
The enrichment acclimation method of above-mentioned thermophilic double oxygen bacterium UTM601 is: the pliotherm period, sludge composting sample 10g joined in the sterilized water of 90ml, on the shaking table of 200rpm, shook 30 minutes, took out then and left standstill 30 minutes; Get the 10ml supernatant and receive in the NA nutrient solution of 90ml, place on the shaking bath, under 60 ℃; The speed change acclimation shaking culture: with per 6 hours was a unit time; With 50rpm is speed change unit, progressively increases rotating speed, after reaching 150rpm; In like manner progressively reduce rotating speed, this is an acclimation period; After accomplishing 3 acclimation periods, get the 10ml domestication liquid and be forwarded in the new NA nutrient solution, continue acclimation shaking culture, so go down to posterity 3 times.
The aerobic performance test screening method of above-mentioned thermophilic double oxygen bacterium UTM601 is: inoculate to the 250ml triangular flask of the sterilized 50ml of being equipped with, 100ml, 150ml, 200ml, 250ml NA nutrient solution by inoculum size 5%; Under 60 ℃, leave standstill and cultivated 3 days, detect the OD of nutrient solution
600
The screening method of the degradation characteristic of above-mentioned thermophilic double oxygen bacterium UTM601 is: the bacterial strain that is taken at activation culture on the NA flat board; Point is connected on the degraded substratum; It is abundant under 60 ℃, to be cultured to bacterium colony; The back drips 2% iodine liquid and observes the size that has or not transparent circle and transparent circle, and measures transparent circle diameter (D) and colony diameter (d) with vernier callipers.Resolving power size by formula Up=(D/d) 2 judges that unit is a millimeter (mm), and Up is big more, and the expression resolving power is big more.Upload to be commissioned to train at the degraded substratum and supported for 4 generations, observe the Up situation of per generation bacterium, eliminate the unstable bacterial strain of degradation capability, obtain the big and stable bacterial strain of degradation capability.The prescription of degraded substratum is the 10g peptone, 3g Carnis Bovis seu Bubali cream, 5g NaCl, 2g Zulkovsky starch, 16g agar, 1000ml zero(ppm) water, pH 7.0-7.2.
The invention provides the microbial inoculum that contains thermophilic double oxygen bacterium UTM601.
The invention provides thermophilic double oxygen bacterium UTM601 or the application of its tunning in sludge composting.
The invention provides the application in preparation compost Inoculant of thermophilic double oxygen bacterium UTM601 or its tunning.
The method for preparing the compost Inoculant provided by the invention may further comprise the steps:
1) actication of culture: the bacterial classification inoculation of getting thermophilic double oxygen bacteria strain UTM601 is cultivated 1-3d in substratum under 50-60 ℃ of condition, make the OD of seed liquor
600Be 0.5-1.2;
2) fermentation seed liquid preparation: in liquid nutrient medium, the preparation seed liquor with the seed liquor enlarged culturing, makes fermentation seed liquid again with the activatory bacterial classification inoculation;
3) with wet feed and siccative mix mixture, be 10-25cm with the mixture thickness that tiles, get step 2) fermentation seed liquid and the mixing solutions of molasses splash on the mixture material; Covering mixture material 10-25cm again by the splash mixing solutions of fermentation seed liquid and molasses of preceding method, so repeats; After reaching 50-150cm to mixture bed of material heap, cover the ventilation property coverture, when treating that temperature rises to 55-70 ℃; Every at a distance from 3-6d stirring 1 time; Reduce to envrionment temperature until leavening temperature, stop fermentation, the tunning oven dry is made the compost Inoculant.
Described wet feed is a feces of livestock and poultry.
Described siccative is peanut hull meal, dregs of beans and/or wood sawdust.
Step 2 wherein) described culture of seed liquid condition is that 50-60 ℃ of concussion cultivated, and rotating speed is 50-200r/min, treats the OD of seed liquor
600During for 1-2, stop to cultivate; The cultural method of described fermentation seed liquid is: with seed liquor by volume 5%-20% be inoculated in liquid nutrient medium and enlarge fermentation culture; Culture condition is temperature 50-60 ℃; Interval 4-8h ventilation 10-30min; Air flow volume ratio 0.3 ~ 0.8:1,100-200rpm stirs, and cultivates the OD of 2-6 days fermentation seed liquids
600Between 2-3, stop fermentation, get fermentation seed liquid.
Wherein, the C/N ratio of the said mixture of step 3) is 20-30, and water ratio is 35%-45%.
Wherein, The said fermentation seed liquid of step 3) with the method that the mixing solutions of molasses is splashed on mixture is: the molasses of getting mixture weight 0.5%-1% are by mass ratio 1:2 and step 2) fermentation seed liquid that makes mixes; Press mass ratio 1:3 again and mix, then mixing solutions is evenly splashed on mixture with water.
In an embodiment of the present invention, the substratum that uses as the NA substratum (Carnis Bovis seu Bubali cream 3-5g, peptone 6-10g, sodium-chlor 3-5g, agar 14-18g, plant gel 7-9g, zero(ppm) water 1000ml, pH7.2).
The present invention also provides the application of compost Inoculant in compost of the above-mentioned UTM601 of containing bacterial strain.
Particularly, the compost Inoculant by weight 10%-20% and mixing of materials, is added auxiliary material, making the mixture C/N ratio is 20-30, and water ratio 50-60% carries out compost.
The present invention from the sludge composting sample that becomes thoroughly decomposed separation screening to a kind of thermophilic double oxygen bacterial strain; This bacterial strain UTM601 equal well-grown in 50-70 ℃ little oxygen and aerobic environment; Have the compost pliotherm period environment of adaptive temperature height, oxygen level skewness one, the characteristics of degradation of organic substances matter fast preferably.It is carrier with organic waste that the present invention utilizes this bacterial strain, prepares solid-state mikrobe compost Inoculant, and cost is low, can not exert an influence to material moisture when being applied to compost.Utilize solid-state mikrobe compost Inoculant of the present invention to carry out During High-Temperature Composting, can under lower multilevel ventilation, realize the thermophilic fermentation of material, avoided the ventilating management control problem of complicacy in the composting process, have simple to operate, energy-conservation characteristics.
Embodiment
Following examples are used to explain the present invention, but are not used for limiting scope of the present invention.Under the situation that does not deviate from the present invention's spirit and essence, modification or replacement to the inventive method, step or condition are done all belong to scope of the present invention.
If do not specialize the conventional means that used technique means is well known to those skilled in the art among the embodiment.
Separation and the evaluation of embodiment 1 thermophilic double oxygen bacterium UTM601
Take by weighing 10g pliotherm period sludge composting sample (picking up from the green source of the earth environmental protection science and technology (Beijing) ltd compost fermentation workshop) and place the 250ml triangular flask that 10 granulated glass spherees and 90ml sterilized water are housed, on the shaking table of 200rpm, shook 30 minutes, take out then and left standstill 30 minutes; Get the 10ml supernatant and receive the NA nutrient solution of 90ml (Carnis Bovis seu Bubali cream 4g, peptone 8g, sodium-chlor 4g; Agar 16g, plant gel 8g, zero(ppm) water 1000ml; PH7.2) in, place on the shaking bath, under 60 ℃; The speed change acclimation shaking culture: with per 6 hours be a unit time, be speed change unit with 50rpm, progressively increase rotating speed; After reaching 150rpm, in like manner progressively reduce rotating speed, this is an acclimation period; After accomplishing 3 acclimation periods, get the 10ml domestication liquid and be forwarded in the new NA nutrient solution, continue acclimation shaking culture, so go down to posterity 3 times.
Get the bacteria suspension 1mL after the domestication respectively, join in the 9mL sterilized water, process 10 with the gradient dilution method
-1, 10
-2, 10
-3, 10
-4, 10
-5, 10
-6, 10
-7Extent of dilution, get the dull and stereotyped coating of 0.1mL, each extent of dilution is coated on 3 NA culture medium flat plates.Get the sterilized water of 1mL and do above-mentioned same operation, as control treatment.Cultivated 2 days down at 60 ℃, select bacterium colony to disperse dull and stereotyped (flat board under certain extent of dilution) preferably, the eugonic single bacterium colony of picking is on the NA substratum; The separation and purification 3 times of ruling repeatedly, again that form is consistent bacterium colony film-making is simply dyeed, observation thalli morphology under oily mirror; The somatic cells form is observed in many visuals field down, after the affirmation thalli morphology unanimity, is seeded to the LB slant culture to abundant; Get the thermophilus strain of purifying, preserve down at 4 ℃, subsequent use.
Get the thermophilus strain of above-mentioned purifying, activation culture is to OD in the NA liquid nutrient medium
600Be 0.8-1.0,5% inoculum size is inoculated to the 250ml triangular flask of the sterilized 50ml of being equipped with, 100ml, 150ml, 200ml, 250ml NA nutrient solution by volume then, under 60 ℃, leaves standstill to cultivate 3 days, detects the OD of nutrient solution
600Decompose the OD of each bacterium under different liquid amounts
600, select OD
600Big and receive liquid amount to influence little bacterial strain, as the thermophilic double oxygen bacterial strain of target.
Get above-mentioned thermophilic double oxygen bacterial strain through after 36 hours, putting and be connected to degraded substratum (10g peptone, 3g Carnis Bovis seu Bubali cream in activation culture on the NA flat board; 5g NaCl, 2g Zulkovsky starch, 16g agar; 1000ml zero(ppm) water, pH7.2) on, it is abundant under 60 ℃, to be cultured to bacterium colony; The back drips 2% iodine liquid and observes the size that has or not transparent circle and transparent circle, and measures transparent circle diameter (D) and colony diameter (d) with vernier callipers.Resolving power size by formula Up=(D/d) 2 judges that unit is a millimeter (mm), and Up is big more, and the expression resolving power is big more.Upload foster 4 generations of being commissioned to train at the degraded substratum; The Up situation of calculating and analysis bacterium of per generation is eliminated the unstable bacterial strain of degradation capability, obtains the big and stable bacterial strain of degradation capability; Therefrom select and get the maximum bacterial strain of Up value, and be numbered UTM601 as the thermophilic little oxygen degraded strain of target.
UTM601 is at NA substratum (Carnis Bovis seu Bubali cream 3-5g, peptone 6-10g, sodium-chlor 3-5g, agar 14-18g; Plant gel 7-9g, zero(ppm) water 1000ml, pH7.2) upward colony characteristics is: form is irregular; Surface wettability, light yellow, erode shape; Flat, growth is cultivated the following 36 hours about 3mm of colony diameter for 50-70 ℃ rapidly.
DNA with UTM601 is a template, its 16S rRNA sequence of pcr amplification, primer is (27f): 5 '-AGA GTT TGA TCC TGG CTC AG-3 ' with (1492r): 5 '-GGTTAC CTT GTTACGACT T-3 '.The PCR response procedures is: 95 ℃ of preparatory sex change 5min, and 95 ℃ of sex change 30s, 53 ℃ of annealing 45s, 72 ℃ are extended 90s, 30 circulations, 72 ℃ are extended 10min.Amplified production checks order after its purity of electrophoresis detection, and sequencing result is seen shown in the sequence table SEQ ID No.1.The 16S rRNA gene order that obtains is compared through EzTaxon DB (http://www.eztaxon.org/), and the result shows itself and Anoxybacillus pushchinoensis K-1
TSimilarity is the highest; Be 98.80%,, and be deposited in (address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, China Committee for Culture Collection of Microorganisms common micro-organisms center on March 22nd, 2012 this bacterial strain called after UTM601; Institute of Microorganism, Academia Sinica; Postcode 100101) preservation, classification called after Anoxybacillus pushchinoensis, preserving number is CGMCC No.5929.
The preparation (1) of embodiment 2 compost Inoculants
(1) preparation activation solution: 4 ℃ of 1 on inclined-planes of preservation getting mikrobe UTM601 of the present invention are seeded to 5 sterilized NA liquid medium test tubes that 10ml is housed, and in 55 ℃ water bath with thermostatic control, cultivate and get slant activation bacterium liquid in 1.5 days.The OD of slant activation bacterium liquid
600Be 0.8.
(2) preparation seed liquor: 1 in above-mentioned slant activation bacterium liquid test tube is seeded in the aseptic NA liquid medium of 1L, and concussion is cultivated on shaking bath, 55 ℃ of culture temperature, and oscillation frequency 50rpm cultivates and gets seed liquor after 3 days.The OD of seed liquor
600Be 1.2.
(3) preparation of fermentation seed liquid: above-mentioned seed liquor is seeded in the sterilized fermentor tank by the inoculum size of 5% (v/v), enlarges fermentation culture.The condition that enlarges fermentation culture is: 55 ℃ of temperature, and ventilation in 4 hours is 15 minutes at interval, air flow volume ratio 0.5:1,200rpm stirs, and cultivates to get fermentation seed liquid in 4 days.The OD of fermentation seed liquid
600Be 2.5.
(4) just be wet feed with chicken manure, peanut hull meal is a siccative, with wet feed and siccative mix mixture; Making C/N ratio is 28; Water ratio is 40%, and it is 10cm that mixture is tiled in cement flooring thickness, and the molasses of getting mixture weight 0.5% are by mass ratio 1:2 and step 2) fermentation seed liquid that makes mixes; Press mass ratio 1:3 again and mix, then mixing solutions is evenly splashed on mixture with water.Covering mixture material 10cm again by the splash mixing solutions of fermentation seed liquid and molasses of preceding method, so repeats; After reaching 50cm to mixture bed of material heap, cover straw screen or mat, when treating that temperature rises to 55 ℃; Every at a distance from 6d stirring 1 time; Reduce to envrionment temperature until leavening temperature, stop fermentation, the tunning drying is made solid-state compost Inoculant.
The preparation (2) of embodiment 3 compost Inoculants
(1) preparation activation solution, culture condition is: 4 ℃ of 1 on inclined-planes of preservation getting mikrobe UTM601 of the present invention are seeded to 5 sterilized NA liquid nutrient medium test tubes that 10ml is housed, and cultivate 2 days the OD of slant activation bacterium liquid in 50 ℃ the water bath with thermostatic control
600Be 1.2.
(2) preparation seed liquor, 50 ℃ of culture temperature, oscillation frequency 160rpm cultivates and gets seed liquor, the OD of seed liquor after 4 days
600Be 1.5.
(3) in the preparation of fermentation seed liquid, the inoculum size of above-mentioned seed liquor is 15% (v/v), and the condition that enlarges fermentation culture is: 50 ℃ of temperature, and ventilation in 5 hours is 20 minutes at interval, air flow volume ratio 0.3:1,160rpm stirs, and cultivates to get fermentation seed liquid in 2 days.The OD of fermentation seed liquid
600Be 2.
(4) with the swine excrement be wet feed, dregs of beans is a siccative, with wet feed and siccative mix mixture; Making C/N ratio is 20; Water ratio is 35%, and it is 18cm that mixture is tiled in cement flooring thickness, and the molasses of getting mixture weight 1% are by mass ratio 1:2 and step 2) fermentation seed liquid that makes mixes; Press mass ratio 1:3 again and mix, then mixing solutions is evenly splashed on mixture with water.Covering mixture material 18cm again by the splash mixing solutions of fermentation seed liquid and molasses of preceding method, so repeats; After reaching 90cm to mixture bed of material heap, cover straw mat, when treating that temperature rises to 62 ℃; Every at a distance from 5d stirring 1 time; Reduce to envrionment temperature until leavening temperature, stop fermentation, the tunning drying is made solid-state compost Inoculant.
The preparation (3) of embodiment 4 compost Inoculants
(1) preparation activation solution, culture condition is: 4 ℃ of 1 on inclined-planes of preservation getting mikrobe UTM601 of the present invention are seeded to 5 sterilized NA liquid medium test tubes that 10ml is housed, and cultivate 1 day the OD of slant activation bacterium liquid in 60 ℃ the water bath with thermostatic control
600Be 0.5.
(2) preparation seed liquor, 60 ℃ of culture temperature, oscillation frequency 200rpm cultivates and gets seed liquor, the OD of seed liquor after 2 days
600Be 1.
(3) in the preparation of fermentation seed liquid, the inoculum size of above-mentioned seed liquor is 20% (v/v), and the condition that enlarges fermentation culture is: 60 ℃ of temperature, and ventilation in 8 hours is 30 minutes at interval, air flow volume ratio 0.8:1,100rpm stirs, and cultivates to get fermentation seed liquid in 6 days.The OD of fermentation seed liquid
600Be 3.
(4) just be wet feed with cow dung, the wood sawdust powder is a siccative, with wet feed and siccative mix mixture; Making C/N ratio is 30; Water ratio is 45%, and it is 25cm that mixture is tiled in cement flooring thickness, and the molasses of getting mixture weight 0.8% are by mass ratio 1:2 and step 2) fermentation seed liquid that makes mixes; Press mass ratio 1:3 again and mix, then mixing solutions is evenly splashed on mixture with water.Covering mixture material 25cm again by the splash mixing solutions of fermentation seed liquid and molasses of preceding method, so repeats; After reaching 150cm to mixture bed of material heap, cover the ventilation property coverture, when treating that temperature rises to 70 ℃; Every at a distance from 3d stirring 1 time; Reduce to envrionment temperature until leavening temperature, stop fermentation, the tunning drying is made solid-state compost Inoculant.
The application of embodiment 5 compost Inoculants
Get compost Inoculant and 60 tons of raw sludges, 20 tons of laggard windrows of peanut hull meal thorough mixing of 10 tons of embodiment, 2 preparations and put fermentation.Be contrast not add compost Inoculant of the present invention simultaneously, experimentize.The compost time is 30 days, during every day detect heap temperature, at interval turning in 5 days once, turning when temperature to be piled reaches 65 ℃; After the principle sampling and abundant mixing by 5 point sampling methods, detected the compost maturity index in being positioned at heap body surface layer 15 ㎝ place in per 5 days, sample also detected physical and chemical indexs such as its nutrient when compost was finished.Interpretation of result is following:
The compost Inoculant of table 1 embodiment 2 is to the impact analysis of compost temperature
The compost Inoculant Inoculant of table 2 embodiment 2 is to the influence of compost maturity
Table 1 and table 2 show, handles with inoculation group not and compare, and the compost Inoculant of inoculation embodiment 2 not only can obviously improve the compost prior fermentation and heat up; Improve compost temperature, and high-temperature duration is long, the compost reaction rate accelerates; Reduce the phytotoxicity of material; Improve the innoxious degree of material, shorten the compost fermentation cycle, promote the compost quick composting.
Nutrient index behind table 3 compost
Table 3 shows, inoculation is compared with not inoculating, and nutrient content increases, and moisture reduces, and is beneficial to the generation of oven dry granulation of composting production later stage and high-quality and efficient fertilizer.
According to above-mentioned same method, adopt the solid dung Inoculant of embodiment 3 and 4 preparations to carry out the compost application, also obtained above-mentioned similar test effect.Explain that the compost Inoculant that thermophilic double oxygen bacterium UTM601 provided by the invention makes can make the heap body play temperature fast, gets into hot stage rapidly, and keeps long-time high temperature; Help the organic quick degraded and the process of becoming thoroughly decomposed; Reduce the phytotoxicity of material, improve the innoxious degree of material, shorten the compost fermentation cycle; Promote the compost quick composting, improved the quality and the fermentation efficiency of compost.
Claims (10)
1. thermophilic double oxygen bacterium (Anoxybacillus pushchinoensis) bacterial strain UTM601, its deposit number is CGMCC No.5929.
2. described thermophilic double oxygen bacterium UTM601 of claim 1 or the application of its tunning in compost.
3. the described thermophilic double oxygen bacterium UTM601 of claim 1 or its tunning application in preparation compost Inoculant.
4. the compost inoculation microbial inoculum that contains the said thermophilic double oxygen bacterium UTM601 of claim 1.
5. a method for preparing the said compost Inoculant of claim 4 is characterized in that, may further comprise the steps:
1) actication of culture: the weighting profit requires the bacterial classification inoculation of 1 said thermophilic double oxygen bacteria strain UTM601 in substratum, under 50-60 ℃ of condition, cultivates 1-3d, makes the OD of seed liquor
600Be 0.5-1.2;
2) fermentation seed liquid preparation: in liquid nutrient medium, the preparation seed liquor with the seed liquor enlarged culturing, makes fermentation seed liquid again with the activatory bacterial classification inoculation;
3) with wet feed and siccative mix mixture, be 10-25cm with the mixture thickness that tiles, get step 2) fermentation seed liquid and the mixing solutions of molasses splash on the mixture material; Covering mixture material 10-25cm again by the splash mixing solutions of fermentation seed liquid and molasses of preceding method, so repeats; After reaching 50-150cm to mixture bed of material heap, cover the ventilation property coverture, when treating that temperature rises to 55-70 ℃; Every at a distance from 3-6d stirring 1 time; Reduce to envrionment temperature until leavening temperature, stop fermentation, the tunning oven dry is made the compost Inoculant.
6. method as claimed in claim 5 is characterized in that step 2) described culture of seed liquid condition is that 50-60 ℃ of concussion cultivated, rotating speed is 50-200r/min, treats the OD of seed liquor
600During for 1-2, stop to cultivate; The cultural method of described fermentation seed liquid is: with seed liquor by volume 5%-20% be inoculated in liquid nutrient medium and enlarge fermentation culture; Culture condition is temperature 50-60 ℃; Interval 4-8h ventilation 10-30min; Air flow volume ratio 0.3 ~ 0.8:1,100-200rpm stirs, and cultivates the OD of 2-6 days fermentation seed liquids
600Between 2-3, stop fermentation, get fermentation seed liquid.
7. method as claimed in claim 5 is characterized in that, the C/N ratio of the said mixture of step 3) is 20-30, and water ratio is 35%-45%.
8. method as claimed in claim 5; It is characterized in that; The said fermentation seed liquid of step 3) with the method that the mixing solutions of molasses is splashed on mixture is: the molasses of getting mixture weight 0.5%-1% are by mass ratio 1:2 and step 2) fermentation seed liquid that makes mixes; Press mass ratio 1:3 again and mix, then mixing solutions is evenly splashed on mixture with water.
9. the application of the described compost Inoculant of claim 4 in compost.
10. the described application of claim 9 is characterized in that, the compost Inoculant by weight 10%-20% and mixing of materials, is added auxiliary material, and making the mixture C/N ratio is 20-30, and water ratio 50-60% carries out compost.
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| CN103725627A (en) * | 2013-11-28 | 2014-04-16 | 大地绿源环保科技(北京)有限公司 | Method of innocent treatment for dead and eliminative chickens by adopting aerobic composting fermentation technology |
| CN106434456A (en) * | 2016-09-29 | 2017-02-22 | 山西宁邦生物肥业有限公司 | Thermophile bacterium A.rupiensis ZQ2 and application thereof |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103695338A (en) * | 2012-12-26 | 2014-04-02 | 大地绿源环保科技(北京)有限公司 | Compound microbial activator and preparation method and application thereof |
| CN103694010A (en) * | 2012-12-26 | 2014-04-02 | 大地绿源环保科技(北京)有限公司 | Ultrahigh-temperature aerobic fermentation method for sludge and application thereof |
| CN103694010B (en) * | 2012-12-26 | 2016-01-20 | 大地绿源环保科技(北京)有限公司 | A kind of Ultrahigh-temperaturaerobic aerobic fermentation method for sludge and application thereof |
| CN103695338B (en) * | 2012-12-26 | 2016-07-06 | 大地绿源环保科技(北京)有限公司 | A kind of Compound microbial activator and preparation method thereof and application |
| CN103725627A (en) * | 2013-11-28 | 2014-04-16 | 大地绿源环保科技(北京)有限公司 | Method of innocent treatment for dead and eliminative chickens by adopting aerobic composting fermentation technology |
| CN103695337A (en) * | 2013-12-06 | 2014-04-02 | 大地绿源环保科技(北京)有限公司 | Method for treating antibiotic dregs by adopting thermophilic microbial agent |
| CN103695337B (en) * | 2013-12-06 | 2015-12-30 | 北京绿源科创环境技术有限公司 | Adopt the method for thermophilic microorganism microbial inoculum process antibiotic bacterium slag |
| CN106434456A (en) * | 2016-09-29 | 2017-02-22 | 山西宁邦生物肥业有限公司 | Thermophile bacterium A.rupiensis ZQ2 and application thereof |
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