CN102732465B - Thermophilic facultative aerobic bacteria UTM601 and application thereof - Google Patents

Thermophilic facultative aerobic bacteria UTM601 and application thereof Download PDF

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CN102732465B
CN102732465B CN2012102269672A CN201210226967A CN102732465B CN 102732465 B CN102732465 B CN 102732465B CN 2012102269672 A CN2012102269672 A CN 2012102269672A CN 201210226967 A CN201210226967 A CN 201210226967A CN 102732465 B CN102732465 B CN 102732465B
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CN102732465A (en
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刘永跃
周顺桂
许宜北
汪涌
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DADI LVYUAN ENVIRONMENTAL PROTECTION TECHNOLOGY (BEIJING) CO LTD
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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

One strain is thermophilic double oxygen bacterium UTM601 and application thereof
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 to make organism mineral, humify and the innoxious process that becomes the fertilizer that becomes thoroughly decomposed by thermophilic fermentation under microbial process.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 technology 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, different sites for different batch mixing ratios, heap body, the range of influence 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 is just so that be difficult to hold suitable ventilation in actually operating: ventilation is too small, microorganism can't obtain enough oxygen, activity decreased, 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 affects 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 is subject to the inhibition of temperature.
Thermophilic double oxygen bacterium has thermophilic, amphimicrobian, than mesophilic bacteria, have advantages of the active and metabolic rate faster of higher organic matter degradation, can improve the degradation rate of organic substance, shorten the process operation time, utilize at recycling organic waste to have 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 yet there are no the open report that is applied to During High-Temperature Composting about thermophilic double oxygen bacterium as Inoculant.
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 in the green source of derived from ground environmental protection science and technology (Beijing) company limited 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 And Nomenclature is Anoxybacillus pushchinoensis, preserving number CGMCC No.5929.
Bacterial strain UTM601 of the present invention has following characteristic:
UTM601 is at NA substratum (extractum carnis 3-5g, peptone 6-10g, sodium-chlor 3-5g, agar 14-18g, plant gel 7-9g, distilled water 1000ml, pH7.2) upper colony characteristics is: form is irregular, surface wettability, light yellow, erode shape, flat, growth is rapidly cultivated approximately 3mm of lower 36 hours colony diameters for 50-70 ℃.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 database (http://www.eztaxon.org/) and carries out sequence analysis, 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 fast degradation starch for this bacterial strain of discriminating under little oxygen condition.
The acclimating method of above-mentioned thermophilic double oxygen bacterium UTM601 is: the pliotherm period, sludge composting sample 10g joined in the sterilized water of 90ml, shaking table at 200rpm shook 30 minutes, then take out and left standstill 30 minutes, getting the 10ml supernatant liquor receives in the NA nutrient solution of 90ml, place on the shaking bath, under 60 ℃, the speed change domestication is cultivated: take per 6 hours as a time unit, take 50rpm as speed change unit, progressively increase rotating speed, until after reaching 150rpm, in like manner progressively reduce rotating speed, this is an acclimation period; After finishing 3 acclimation periods, get the 10ml domestication liquid and be forwarded in the new NA nutrient solution, continue domestication and cultivate, so go down to posterity 3 times.
The aerobic performance test screen 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, under 60 ℃, be cultured to bacterium colony abundant, rear dropping 2% iodine liquid is observed the size that has or not transparent circle and transparent circle, and measures transparent circle diameter (D) and colony diameter (d) with vernier callipers.The capacity of decomposition size is Up=(D/d by formula) 2 to judge, unit is millimeter (mm), and Up is larger, and the expression capacity of decomposition is larger.Upload 4 generations of culture at the degraded substratum, observe the Up situation of per generation bacterium, eliminate the unstable bacterial strain of degradation capability, obtain the large and stable bacterial strain of degradation capability.The prescription of degraded substratum is the 10g peptone, 3g extractum carnis, 5g NaCl, 2g Zulkovsky starch, 16g agar, 1000ml distilled 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: get the bacterial classification inoculation of thermophilic double oxygen bacteria strain UTM601 in substratum, under 50-60 ℃ of condition, cultivate 1-3d, 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 bacterial classification inoculation of activation;
3) wet feed and siccative are mixed to get 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 repeat, after reaching 50-150cm to mixture bed of material heap, cover the ventilation property coverture, when treating that temperature rises to 55-70 ℃, stir 1 time every 3-6d, until leavening temperature is down to envrionment temperature, stop fermentation, the tunning oven dry is made the compost Inoculant.
Described wet feed is feces of livestock and poultry.
Described siccative is peanut hull meal, dregs of beans and/or wood sawdust.
Step 2 wherein) culture condition of described seed liquor 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 ℃, the interval 4-8h 10-30min that ventilates, 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 carbon-nitrogen ratio of the described mixture of step 3) is 20-30, and water ratio is 35%-45%.
Wherein, the described 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 1:2 and step 2 in mass ratio) fermentation seed liquid that makes mixes, 1:3 mixes with water in mass ratio again, then mixing solutions is evenly splashed on mixture.
In an embodiment of the present invention, the substratum that uses is NA substratum (extractum carnis 3-5g, peptone 6-10g, sodium-chlor 3-5g, agar 14-18g, plant gel 7-9g, distilled 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 carbon-nitrogen 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 little oxygen of 50-70 ℃ and aerobic environment, have that adaptive temperature preferably is high, the compost pliotherm period environment of oxygen level distributing inhomogeneity, the characteristics of fast degradation organic substance.The present invention utilizes this bacterial strain take organic waste as carrier, preparation solid-state microorganism compost Inoculant, and cost is low, can not exert an influence to material moisture when being applied to compost.Utilize solid-state microorganism 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 complexity in the composting process, have simple to operate, energy-conservation characteristics.
Embodiment
Following examples are used for explanation the present invention, but are not used for limiting the scope of the invention.Without departing from the spirit and substance of the case in the present invention, 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 (the green source of derived from ground environmental protection science and technology (Beijing) company limited compost fermentation workshop) and place the 250ml triangular flask that 10 granulated glass spherees and 90ml sterilized water are housed, shaking table at 200rpm shook 30 minutes, then take out and left standstill 30 minutes, get the 10ml supernatant liquor and receive NA nutrient solution (the extractum carnis 4g of 90ml, peptone 8g, sodium-chlor 4g, agar 16g, plant gel 8g, distilled water 1000ml, pH7.2) in, place on the shaking bath, under 60 ℃, the speed change domestication is cultivated: take per 6 hours as a time unit, take 50rpm as speed change unit, progressively increase rotating speed, until after reaching 150rpm, in like manner progressively reduce rotating speed, this is an acclimation period; After finishing 3 acclimation periods, get the 10ml domestication liquid and be forwarded in the new NA nutrient solution, continue domestication and cultivate, so go down to posterity 3 times.
Get respectively the bacteria suspension 1mL after the domestication, join in the 9mL sterilized water, make 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, process in contrast.60 ℃ of lower cultivations 2 days, select bacterium colony to disperse preferably dull and stereotyped (flat board under certain extent of dilution), the eugonic single bacterium colony of picking, on the NA substratum, the separation and purification 3 times of repeatedly ruling, again the bacterium colony film-making that form is consistent, simply dye, at oily Microscopic observation thalli morphology, observe the somatic cells form under many visuals field, after the affirmation thalli morphology is consistent, be seeded to the LB slant culture to abundant, get the thermophilus strain of purifying, 4 ℃ of lower preservations, for 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, then 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, leaves standstill to cultivate 3 days under 60 ℃, detects the OD of nutrient solution 600Decompose the OD of each bacterium under different liquid amounts 600, select OD 600Large and be subjected to liquid amount to affect little bacterial strain, as the thermophilic double oxygen bacterial strain of target.
Get above-mentioned thermophilic double oxygen bacterial strain through in activation culture on the NA flat board after 36 hours, point is connected to degraded substratum (10g peptone, the 3g extractum carnis, 5g NaCl, 2g Zulkovsky starch, 16g agar, 1000ml distilled water, pH7.2) on, under 60 ℃, be cultured to bacterium colony abundant, rear dropping 2% iodine liquid is observed the size that has or not transparent circle and transparent circle, and measures transparent circle diameter (D) and colony diameter (d) with vernier callipers.The capacity of decomposition size is Up=(D/d by formula) 2 to judge, unit is millimeter (mm), and Up is larger, and the expression capacity of decomposition is larger.Upload 4 generations of culture 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 large and stable bacterial strain of degradation capability, therefrom select and get the maximum bacterial strain of Up value as the thermophilic little oxygen degraded strain of target, and be numbered UTM601.
UTM601 is at NA substratum (extractum carnis 3-5g, peptone 6-10g, sodium-chlor 3-5g, agar 14-18g, plant gel 7-9g, distilled water 1000ml, pH7.2) upper colony characteristics is: form is irregular, surface wettability, light yellow, erode shape, flat, growth is rapidly cultivated approximately 3mm of lower 36 hours colony diameters for 50-70 ℃.
Take the DNA of UTM601 as template, its 16S rRNA sequence of pcr amplification, primer is (27f): 5 '-AGA GTT TGA TCC TGG CTC AG-3 ' and (1492r): 5 '-GGTTAC CTT GTTACGACT T-3 '.The PCR response procedures is: 95 ℃ of denaturation 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 by EzTaxon database (http://www.eztaxon.org/), the result shows that itself and Anoxybacillus pushchinoensis K-1T similarity are the highest, be 98.80%, this bacterial strain called after UTM601, 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, Institute of Microorganism, Academia Sinica, postcode 100101) preservation, Classification And Nomenclature is Anoxybacillus pushchinoensis, and 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 microorganism UTM601 of the present invention are seeded to 5 sterilized NA liquid medium test tubes that 10ml is housed, and cultivate 1.5 days to get slant activation bacterium liquid in 55 ℃ water bath with thermostatic control.The OD600 of slant activation bacterium liquid is 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, cultivates in the shaking bath concussion, and 55 ℃ of culture temperature, oscillation frequency 50rpm cultivates and gets seed liquor after 3 days.The OD600 of seed liquor is 1.2.
(3) preparation of fermentation seed liquid: above-mentioned seed liquor is by 5%(v/v) inoculum size be seeded in the sterilized fermentor tank, enlarge fermentation culture.The condition that enlarges fermentation culture is: 55 ℃ of temperature, and interval ventilation in 4 hours 15 minutes, air flow volume ratio 0.5:1,200rpm stirs, and cultivates 4 days to get fermentation seed liquid.The OD600 of fermentation seed liquid is 2.5.
(4) take chicken manure just as wet feed, peanut hull meal is siccative, wet feed and siccative are mixed to get mixture, making carbon-nitrogen 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 1:2 and step 2 in mass ratio) fermentation seed liquid that makes mixes, 1:3 mixes with water in mass ratio again, then mixing solutions is evenly splashed on mixture.Covering mixture material 10cm again, by the splash mixing solutions of fermentation seed liquid and molasses of preceding method, so repeat, after reaching 50cm to mixture bed of material heap, cover straw screen or mat, when treating that temperature rises to 55 ℃, stir 1 time every 6d, until leavening temperature is down to envrionment 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 microorganism 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), the condition that enlarges fermentation culture is: temperature 50 C, interval ventilation in 5 hours 20 minutes, air flow volume ratio 0.3:1,160rpm stirs, and cultivates 2 days to get fermentation seed liquid.The OD of fermentation seed liquid 600Be 2.
(4) take swine excrement as wet feed, dregs of beans is siccative, wet feed and siccative are mixed to get mixture, making carbon-nitrogen 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 1:2 and step 2 in mass ratio) fermentation seed liquid that makes mixes, 1:3 mixes with water in mass ratio again, then mixing solutions is evenly splashed on mixture.Covering mixture material 18cm again, by the splash mixing solutions of fermentation seed liquid and molasses of preceding method, so repeat, after reaching 90cm to mixture bed of material heap, cover straw mat, when treating that temperature rises to 62 ℃, stir 1 time every 5d, until leavening temperature is down to envrionment 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 microorganism 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), the condition that enlarges fermentation culture is: temperature 60 C, interval ventilation in 8 hours 30 minutes, air flow volume ratio 0.8:1,100rpm stirs, and cultivates 6 days to get fermentation seed liquid.The OD of fermentation seed liquid 600Be 3.
(4) take cow dung just as wet feed, the wood sawdust powder is siccative, wet feed and siccative are mixed to get mixture, making carbon-nitrogen 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 1:2 and step 2 in mass ratio) fermentation seed liquid that makes mixes, 1:3 mixes with water in mass ratio again, then mixing solutions is evenly splashed on mixture.Covering mixture material 25cm again, by the splash mixing solutions of fermentation seed liquid and molasses of preceding method, so repeat, after reaching 150cm to mixture bed of material heap, cover the ventilation property coverture, when treating that temperature rises to 70 ℃, stir 1 time every 3d, until leavening temperature is down to envrionment temperature, stop fermentation, the tunning drying is made solid-state compost Inoculant.
The application of embodiment 5 compost Inoculants
The compost Inoculant of getting 10 tons of embodiment, 2 preparations fully mixes laggard windrow and puts fermentation with 60 tons of raw sludges, 20 tons of peanut hull meals.Not add compost Inoculant of the present invention as contrast, test simultaneously.The compost time is 30 days, during every day detect heap temperature, 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 surface layer 15 ㎝ places in per 5 days, sample also detected the physical and chemical indexs such as its nutrient when compost was finished.Interpretation of result is as follows:
The compost Inoculant of table 1 embodiment 2 is to the impact analysis of compost temperature
Figure BDA00001834461200091
The compost Inoculant Inoculant of table 2 embodiment 2 is on the impact of compost maturity
Figure BDA00001834461200101
Table 1 and table 2 show, process with inoculation group not and to compare, 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 behind table 3 compost
Figure BDA00001834461200102
Table 3 shows, inoculation is compared with not inoculating, and nutrient content increases, and moisture reduces, and is beneficial to the generation of the 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.Illustrate that the compost Inoculant that thermophilic double oxygen bacterium UTM601 provided by the invention makes can make the heap body play fast temperature, enter rapidly hot stage, and keep long-time high temperature, be conducive to organic fast degradation and maturity, reduce the phytotoxicity of material, improve the innoxious degree of material, shorten the compost fermentation cycle, promote the compost quick composting, improved quality and the fermentation efficiency of compost.
Figure IDA00001834462100011
Figure IDA00001834462100021

Claims (10)

1. Pu Xinuo anaerobic spore-bearing bacilli (Anoxybacillus pushchinoensis) bacterial strain UTM601, its deposit number is CGMCC No.5929.
2. Pu Xinuo anaerobic spore-bearing bacilli UTM601 claimed in claim 1 or the application of its tunning in compost.
3. Pu Xinuo anaerobic spore-bearing bacilli UTM601 claimed in claim 1 or its tunning application in preparation compost Inoculant.
4. the compost Inoculant that contains the described Pu Xinuo anaerobic spore-bearing bacilli of claim 1 UTM601.
5. a method for preparing the described 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 described Pu Xinuo anaerobic spore-bearing bacilli bacterial strain UTM601 in substratum, cultivates 1-3d under 50-60 ℃ of condition, 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 bacterial classification inoculation of activation;
3) wet feed and siccative are mixed to get 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 repeat, after reaching 50-150cm to mixture bed of material heap, cover the ventilation property coverture, when treating that temperature rises to 55-70 ℃, stir 1 time every 3-6d, until leavening temperature is down to envrionment 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) culture condition of described seed liquor 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 ℃, the interval 4-8h 10-30min that ventilates, 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 carbon-nitrogen ratio of the described 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 described 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 1:2 and step 2 in mass ratio) fermentation seed liquid that makes mixes, 1:3 mixes with water in mass ratio again, then mixing solutions is evenly splashed on mixture.
9. the application of compost Inoculant claimed in claim 4 in compost.
10. application claimed in 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 carbon-nitrogen ratio is 20-30, and water ratio 50-60% carries out compost.
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