CN103497915A - Complex microbial agent for composting fermentation and application thereof - Google Patents
Complex microbial agent for composting fermentation and application thereof Download PDFInfo
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- CN103497915A CN103497915A CN201310430957.5A CN201310430957A CN103497915A CN 103497915 A CN103497915 A CN 103497915A CN 201310430957 A CN201310430957 A CN 201310430957A CN 103497915 A CN103497915 A CN 103497915A
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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
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
- Y02A40/20—Fertilizers of biological origin, e.g. guano or fertilizers made from animal corpses
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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
Abstract
The invention discloses a complex microbial agent for composting fermentation and application thereof. The complex microbial agent is a solid inoculant which comprises phanerochaete chrysosporium, streptomyces griseorubens, bacillus subtilis, bacillus methylotrophicus and bacillus amyloliquefaciens. The complex microbial agent is a complex microbial system comprising various microorganisms which have complementary and synergistic functions; the microbial system integrates quick fermentation heat production, biological deodorization and compost material decomposition and degradation and has the effects of a biological fertilizer; the livestock manure compost can be quickly heated to kill off harmful organisms at high temperature; the unpleasant smell is obviously reduced to solve the problem of environment pollution caused by poultry excrement; the microorganisms can be quickly propagated in the poultry excrement fermentation process; the organic matters such as lignocellulose in the excrement can be quickly and effectively decomposed so as to increase the decomposition speed of the compost material and improve the quality of the compost product; the complex microbial agent can be used for producing the biological organic fertilizer so as to reduce the agricultural production cost.
Description
Technical field
The invention belongs to biological fermentation and microorganism field, apply for compost fermentation complex bacterial agent and tool in particular to a kind of.
Background technology
Along with the quick intensivization development of milk cattle cultivating industry, the improper processing pollution on the environment of cow manure is also more and more serious.At present, most raisers impose on the direct as fertilizer sources of cow dung farmland, concentrate on air storage or pour pond, ditch etc., cause the fecal pollution problem day by day to manifest.From the angle of environment, it has caused pollution to atmosphere, soil, waters, and ecotope and resident's living environment is worsened, and is detrimental to health, and also is unfavorable for the healthy growth of milk cow simultaneously.But the angle from resource, it is but good renewable resources, if scientific utilization, it can not only solve shortage of resources and this two hang-up of environmental pollution, can also be for green agriculture, pollution-less agriculture production provide safeguard, for agricultural sustainable development is offered reference.
That During High-Temperature Composting has is with low cost, can effectively kill pathogenic bacteria, improve the advantage such as physical behavior, non-secondary pollution of animal waste, is to be widely used at present and the cow dung processing mode of tool prospect.Yet cow dung is of close texture, water content is high, air permeability is poor, leavening temperature is low, belongs to cold manure; In addition, robust fibre in cow dung accounts for the 30-50% of dry weight, lignocellulose in compost amendment (agricultural crop straw) accounts for the 60-80% of dry weight, wherein Mierocrystalline cellulose and hemicellulose are macromolecular polysaccharides, and the two is can be under microbial process relatively easy resolves into the carbohydrate that can be utilized by organism; And xylogen is poly-phenols polymer aromatics, wherein contain a large amount of benzene ring structures, cause its degraded very difficult.And xylogen, Mierocrystalline cellulose and hemicellulose mutually intert, combine closely on space structure, greatly reduce its biological degradability.Many results of study are verified, and the degraded of lignocellulose is restriction compost maturity process the key factor that affects quality of compost products.
For the cow dung compost slow and lignocellulose composition difficult problem of degrading wherein that heats up, usually adopt to the method that adds functional microorganism in compost and accelerate the maturity of compost and improve the quality of composting production.But current ubiquitous problem is in complex microorganism screening and culturing process, lacks basic theoretical direction, cause the bacterial strain that filters out functional not strong; And, comparatively not familiar to the fermenting process of compound cultivation and the interaction between each bacterial strain thereof and mechanism thereof, cause the product total viable count not high, the ideal effect that compost is not reached use composite fungus agent to promote maturity.
Summary of the invention
The object of the present invention is to provide a kind of microbiobacterial agent and application thereof that the cow dung compost of making is rapidly heated and stronger ligocellulose degradation ability is arranged that have.In order to realize purpose of the present invention, intend adopting following technical scheme:
Aspect of the present invention relates to a kind of composite fungus agent, it is characterized in that omiting the weight ratio preparation of the solid fungicide of red streptomyces, subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens according to 1-2:1-2:1-2:1-2:1-2 by Phanerochaete chrysosporium, ash.
In another aspect of this invention, also relate to the preparation method of above-mentioned composite fungus agent, it is characterized in that comprising the steps:
(1) slant culture: the original strain that Phanerochaete chrysosporium, ash is omited to red streptomyces, subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens is inoculated in respectively on corresponding solid medium, cultivate 3-5 days by Phanerochaete chrysosporium 28-30 ℃, ash is omited to 28-30 ℃ of red streptomyces and cultivate 5-7 days, by subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens 30-37 ℃ of cultivation 2-3 days, make actication of culture;
(2) first order seed is cultivated: with stroke-physiological saline solution washing activated inclined plane to shaking culture in aseptic triangular flask; Subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens are cultivated 40-60h at 28-32 ℃, 150-200r/min and are obtained primary seed solution; Phanerochaete chrysosporium is cultivated 70-80h at 26-30 ℃, 200-240r/min and obtains primary seed solution; Ash slightly red streptomyces is cultivated 90-120h and is obtained primary seed solution at 26-30 ℃, 200-240r/min;
(3) secondary seed is cultivated: the inoculum size that is 10-20% by the volume ratio of liquid nutrient medium, first order seed is inoculated in respectively in fermentor tank, and cultivate 1-3 days under subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens 30-37 ℃ condition; Phanerochaete chrysosporium 25-30 ℃, stirring velocity are 100-200r/min, air flow is 1:0.5-1, cultivates 4-6 days: ash slightly red streptomyces is 150-220r/min at 28-30 ℃, stirring velocity, and air flow is 1:1-1.5, cultivate 4-6 days, make secondary seed;
(4) the bacterial classification solid fermentation is cultivated: the secondary seed fermented liquid of each bacterial strain and sterilizing wheat bran are dried after the ratio mixed fermentation 24-72h with 1:1, make solid fungicide;
(5) composite fungus agent preparation: the solid fungicide that Phanerochaete chrysosporium, ash is omited to red streptomyces, subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens mixes according to part by weight, makes composite fungus agent.
The present invention also relates to the application of above-mentioned composite fungus agent on the other hand, and described composite fungus agent is for cow dung compost, preferred, for reducing the content of lignin in compost.
In a preferred embodiment of the present invention, described cow dung compost comprises the steps:
(1) composting material is fresh cow dung and rice straw, rice straw is cut up with a hay cutter into to the segment of 3-8cm, with the mass ratio of bright cow dung according to 1:3-4, mixes, and regulates moisture content to 60～65%, piles fermentation heap;
(2) microbial inoculum is added on Jian Dui limit in limit, and the amount of described microbial inoculum is 5-15 * 10
9cFUkg
-1composting material, even hybrid reactor body material after inoculation; Turning when temperature rises to more than 45 ℃, every 3d turning 1 time, after temperature drops to below 35 ℃, every 7d turns over 1 time, until heap temperature drops to envrionment temperature, finishes fermentation.
Composite fungus agent of the present invention is the living multiple-microorganism composite microbial system of association that has complementary functions, the heat production of fungus strain collection quick fermentation, biological deodorizing, compost material becomes thoroughly decomposed and degrades in one, and have the effect of bio-feritlizer concurrently, can make livestock excrement composting be rapidly heated, at high temperature kill harmful organism, obviously reduce unhappy smell, the environmental pollution caused to solve feces of livestock and poultry, can in the livestock manure fermented process, breed rapidly by microorganism, decompose fast effectively in ight soil the organic substances such as lignocellulose, thereby accelerate the speed of becoming thoroughly decomposed of compost material, promote quality of compost products, can be used for producing biological organic fertilizer, thereby reduction agriculture production cost.
The accompanying drawing explanation
The variation of temperature in Fig. 1: embodiment 1 Composting Process;
The variation of temperature in Fig. 2: embodiment 2 Composting Process;
The variation of temperature in Fig. 3: embodiment 3 Composting Process.
Embodiment
In order further to set forth related material and the technique of technical scheme of the present invention, following examples have been provided.The scope that these embodiment do not limit the present invention in any way.
Embodiment 1:
1.1 microbiobacterial agent preparation
1.1.1 slant culture: by the original strain of Phanerochaete chrysosporium, grey slightly red streptomyces, subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens, (above-mentioned bacterial classification is the common bacterial classification in this area, and can be commercially available) be inoculated in respectively on corresponding solid medium, cultivate 3-5 days by Phanerochaete chrysosporium 28-30 ℃, ash is omited to 28-30 ℃ of red streptomyces and cultivate 5-7 days, by subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens 30-37 ℃ of cultivation 2-3 days, make actication of culture.
1.1.2 first order seed is cultivated: with stroke-physiological saline solution washing activated inclined plane to shaking culture in aseptic triangular flask.Subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens are cultivated 48h at 30 ℃, 180r/mmin and are obtained primary seed solution;
Phanerochaete chrysosporium is cultivated 72h at 28 ℃, 220r/min and obtains primary seed solution; Ash slightly red streptomyces is cultivated 96h and is obtained primary seed solution at 28 ℃, 220r/mmin.
1.1.3 secondary seed is cultivated: the inoculum size that is 10-20% by the volume ratio of liquid nutrient medium, first order seed is inoculated in respectively in fermentor tank, cultivate 1-3 days under subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens 30-37 ℃ condition; Phanerochaete chrysosporium 25-30 ℃, stirring velocity are 100-200r/min, and air flow is 1:0.5-1, cultivate 4-6 days; Ash slightly red streptomyces is 150-220r/min at 28-30 ℃, stirring velocity, and air flow is 1:1-1.5, cultivates 4-6 days, makes secondary seed.
1.1.4 the bacterial classification solid fermentation is cultivated: the secondary seed fermented liquid of each bacterial strain and sterilizing wheat bran are dried after the ratio mixed fermentation 24-72h with 1:1, make solid fungicide.
1.1.5 composite fungus agent preparation: the solid fungicide that Phanerochaete chrysosporium, ash is omited to red streptomyces, subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens mixes according to the ratio of 1:1:1:1:1:1, makes composite fungus agent standby.
1.2 composting material
Composting material is fresh cow dung and rice straw (amendment).Rice straw is cut up with a hay cutter into to the segment of 5cm left and right, mix with the mass ratio of bright cow dung according to 1:3.5, regulate moisture content to 60～65%, pile the fermentation heap of 1m * 1m * 1.2m.This windrow organic content is about 59.5%, C/N and is about 30:1, and the wood fibre cellulose content is 55.59%, and wherein xylogen, Mierocrystalline cellulose, hemicellulose ratio are respectively 6.84%, 24.37% and 24.38%.
1.3 compost and the method for sampling
Windrow divides A, B group, and the A group, for control sample, is carried out Natural compost; The B group is added this composite fungus agent.Adopt solid-state inoculum, concentration is 1 * 10
9cFUg
-1, inoculum size is 10gkg
-1, microbial inoculum is added on Jian Dui limit, limit, even hybrid reactor body material after inoculation.Start turnings when temperature rises to 45 ℃, every 3d turning 1 time, temperature after dropping to 35 ℃ every 7d turn over 1 time, finish fermentation until heap temperature drops to envrionment temperature, obtain sufficient oxygen to guarantee windrow.Compost continues 46d, according to heap temperature, changes sampling analysis.Take the multi-point sampling method, evenly, after biased sample, measure xylogen, Mierocrystalline cellulose and hemicellulose level, all analytical samplings all gather 3 Duplicate Samples.
1.4 compost temperature and lignocellulose are measured
Use LNI-T UT325 digital electronic thermometer to measure envrionment temperature and pile body 30cmm place temperature every day; Utilize ANKOM
220fiber type analysis-e/or determining wood fibre cellulose content.
1.5 experimental result
1.5.1 the variation of temperature in composting process
The compost of inoculation microbial inoculum just enters the pyrolytic decomposition stage (>45 ℃) (seeing Fig. 1) in the 2nd day after composting, and lasts till the 18th day, and temperature descends gradually afterwards, in the time of 22 days, is down to below 35 ℃.In whole composting process, the pliotherm period, (2～18d) maintained 16 days, and accumulated temperature is 706.1 ℃, and compost temperature has reached the basic demand of compost hazard-free
[16].And although the heap temperature of Natural compost fails to enter the pliotherm period within whole experimental period higher than room temperature.The temperature variation of compost has intuitively been reacted the differentiation of heat in the composting system, is also the important indicator of microbial metabolism activity, and visible inoculation is processed and improved in the Composting Process the especially microorganism active in early stage.
1.5.2 the content of lignocellulose in composting process
As shown in Table 1, when compost finishes, in the compost of interpolation microbial inoculum, the degradation rate of Mierocrystalline cellulose, hemicellulose and xylogen has improved 31.31%, 19.57% and 14.33% than contrast respectively, illustrate that in microbial inoculum, lignocellulose degrading bacteria all has promotion in various degree to ligocellulose degradation in compost, thereby accelerate the compost maturity process.
Table 1 composting process ligocellulose degradation leads
Embodiment 2:
Blending ratio when difference from Example 1 is prepared by composite fungus agent is: Phanerochaete chrysosporium, the ash slightly solid fungicide of red streptomyces, subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens mix according to the ratio of 2:2:1:1:1.
Composting material and method, sampling are all identical with embodiment 1 with measuring method.
2.1 experimental result
2.1.1 the variation of temperature in composting process
The compost of inoculation microbial inoculum is equally also just to enter the pyrolytic decomposition stage (>45 ℃) (seeing Fig. 2) in the 2nd day after composting, and the pliotherm period has continued 18 days, than embodiment 1, has extended 2 days.
2.1.2 the content of lignocellulose in composting process
As shown in Table 2, when compost finishes, in the compost of interpolation microbial inoculum, the degradation rate of Mierocrystalline cellulose, hemicellulose and xylogen has improved 32.5%, 22.48% and 16.17% than contrast respectively, illustrate that in microbial inoculum, lignocellulose degrading bacteria all has promotion in various degree to ligocellulose degradation in compost, thereby accelerate the compost maturity process.
Table 2 composting process ligocellulose degradation leads
Embodiment 3:
Blending ratio when difference from Example 1 is prepared by composite fungus agent is: Phanerochaete chrysosporium, the ash slightly solid fungicide of red streptomyces, subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens mix according to the ratio of 2:1:2:1:2.
Composting material and method, sampling are all identical with embodiment 1 with measuring method.
2.1 experimental result
2.1.1 the variation of temperature in composting process
The compost of inoculation microbial inoculum is equally also just to enter the pyrolytic decomposition stage (>45 ℃) (seeing Fig. 3) in the 2nd day after composting, and the pliotherm period has continued 17 days, than embodiment 1, has extended 1 day.
2.1.2 the content of lignocellulose in composting process
As shown in Table 3, when compost finishes, in the compost of interpolation microbial inoculum, the degradation rate of Mierocrystalline cellulose, hemicellulose and xylogen has improved 30.91%, 21.7% and 14.66% than contrast respectively, illustrate that in microbial inoculum, lignocellulose degrading bacteria all has promotion in various degree to ligocellulose degradation in compost, thereby accelerate the compost maturity process.
Table 3 composting process ligocellulose degradation leads
The above, be only the specific embodiment of the present invention, but protection scope of the present invention is not limited to this, and any variation of expecting without creative work or replacement, within all should being encompassed in protection scope of the present invention.Therefore, protection scope of the present invention should be as the criterion with the protection domain that claims were limited.
Claims (5)
1. a composite fungus agent, is characterized in that comprising the slightly solid fungicide of red streptomyces, subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens of Phanerochaete chrysosporium, ash.
2. a composite fungus agent, is characterized in that omiting the weight ratio preparation of the solid fungicide of red streptomyces, subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens according to 1-2:1-2:1-2:1-2:1-2 by Phanerochaete chrysosporium, ash.
3. the preparation method of claim 1 or 2 described composite fungus agent, is characterized in that comprising the steps:
(1) slant culture: the original strain that Phanerochaete chrysosporium, ash is omited to red streptomyces, subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens is inoculated in respectively on corresponding solid medium, cultivate 3-5 days by Phanerochaete chrysosporium 28-30 ℃, ash is omited to 28-30 ℃ of red streptomyces and cultivate 5-7 days, by subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens 30-37 ℃ of cultivation 2-3 days, make actication of culture;
(2) first order seed is cultivated: with stroke-physiological saline solution washing activated inclined plane to shaking culture in aseptic triangular flask; Subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens are cultivated 40-60h at 28-32 ℃, 150-200r/min and are obtained primary seed solution; Phanerochaete chrysosporium is cultivated 70-80h at 26-30 ℃, 200-240r/min and obtains primary seed solution; Ash slightly red streptomyces is cultivated 90-120h and is obtained primary seed solution at 26-30 ℃, 200-240r/mmin;
(3) secondary seed is cultivated: the inoculum size that is 10-20% by the volume ratio of liquid nutrient medium, first order seed is inoculated in respectively in fermentor tank, and cultivate 1-3 days under subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens 30-37 ℃ condition; Phanerochaete chrysosporium 25-30 ℃, stirring velocity are 100-200r/min, and air flow is 1:0.5-1, cultivate 4-6 days; Ash slightly red streptomyces is 150-220r/min at 28-30 ℃, stirring velocity, and air flow is 1:1-1.5, cultivates 4-6 days, makes secondary seed;
(4) the bacterial classification solid fermentation is cultivated: the secondary seed fermented liquid of each bacterial strain and sterilizing wheat bran are dried after the ratio mixed fermentation 24-72h with 1:1, make solid fungicide;
(5) composite fungus agent preparation: the solid fungicide that Phanerochaete chrysosporium, ash is omited to red streptomyces, subtilis, methylotrophy type genus bacillus and bacillus amyloliquefaciens mixes according to part by weight, makes composite fungus agent.
4. the application of claim 1 or 2 described composite fungus agent, described composite fungus agent is for cow dung compost, preferred, for reducing the content of lignin in compost.
5. application according to claim 4, described cow dung compost comprises the steps:
(1) composting material is fresh cow dung and rice straw, rice straw is cut up with a hay cutter into to the segment of 3-8cm, with the mass ratio of bright cow dung according to 1:3-4, mixes, and regulates moisture content to 60～65%, piles fermentation heap;
(2) microbial inoculum is added on Jian Dui limit in limit, and the amount of described microbial inoculum is 5-15 * 10
9cFUkg
-1composting material, even hybrid reactor body material after inoculation; Turning when temperature rises to more than 45 ℃, every 3d turning 1 time, after temperature drops to below 35 ℃, every 7d turns over 1 time, until heap temperature drops to envrionment temperature, finishes fermentation.
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