CN103484396A - New strain of streptomyces thermocarboxydus and application thereof - Google Patents
New strain of streptomyces thermocarboxydus and application thereof Download PDFInfo
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- CN103484396A CN103484396A CN201310356764.XA CN201310356764A CN103484396A CN 103484396 A CN103484396 A CN 103484396A CN 201310356764 A CN201310356764 A CN 201310356764A CN 103484396 A CN103484396 A CN 103484396A
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- carbon monoxide
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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 discloses a new strain CF1 of streptomyces thermocarboxydus, wherein the collection number is CGMCC (China General Microbiological Culture Collection Center) No.6873, the collection date is November 23, 2012, and the collection unit is CGMCC. The invention further discloses a bacterial agent containing the strain. The new strain CF1 of the streptomyces thermocarboxydus, disclosed by the invention, is inoculated into a compost, so that the temperature of the compost can be rapidly raised in low-temperature seasons in the North, the high temperature of above 50 DEG C of the compost can be maintained for 15 days, the fermentation period is shortened, and pathogens and parasites in animal dung can be effectively killed. After the end of fermentation of the compost, the nutrient content in a composted product is high, the pH value is about 8.0, and the composted product can be used as an excellent plant organic fertilizer.
Description
Technical field
The invention belongs to the soil and fertilizer field, be specifically related to a kind of agricultural microorganism preparation and application thereof.
Background technology
Along with socioeconomic high speed development, people's standard of living improves constantly, and huge variation has occurred dietary structure, and wherein animal products in occupation of indispensable vital role, has promoted thus the fast development of livestock and poultry breeding industry in dietary structure.Recent two decades, the plant that a collection of intensive degree is more and more higher sets up in nearly outer suburbs, the city of convenient transportation one after another under the active support of the government, raise quantity many by few change, a large amount of feces of livestock and poultry, because not having timely harmless treatment arbitrarily to be discarded in physical environment, has caused significant wastage and the environmental pollution of resource.Met people to meat, egg, newborn demand in, brought huge pressure to the natural ecological environment of China, cause serious pollution.
Summary of the invention
At first the present invention provides thermophilic carbon monoxide streptomycete (Streptomyces thermocarboxydus) new bacterial strain CF1.
The present invention also provides the microbial inoculum that contains thermophilic carbon monoxide streptomycete CF1.
Strains separation of the present invention and screening process are as follows:
1. adopt plate dilution method to carry out the microorganism separation from fresh cow dung, thermophilic fermentation stage cow dung and in the cow dung becoming thoroughly decomposed respectively.Take each 10g of each sample and put into respectively the triangular flask that the 90mL sterilized water is housed, the 30min that vibrates on shaking table, fully disperse sample, after standing 20min, is 10
-1diluent, then serial dilution makes 10
-2-10
-8the diluent of concentration is standby.Get the diluent of 100 each concentration of μ L, adopt conventional streak method to be coated with on No. 1 solid medium of Gao Shi with spreader, three repetitions of each concentration gradient, be placed in respectively 25 ℃, 30 ℃, 50 ℃ constant incubators and cultivate.The single bacterium colony grown on the picking flat board after 3d, be further purified cultivation, and in 4 ℃ of preservations.
No. 1 substratum of Gao Shi is Zulkovsky starch 20.0g/L, KNO
31.0g/L, K
2hPO
40.5g/L, MgSO
47H
2o 0.5g/L, NaCl 0.5g/L, FeSO
40.01g/L, water 1000ml, pH7.2-7.4, agar 1.5%-2.0%.
2. form the size of the distinct degree of speed, transparent circle, transparent circle and the colony diameter ratio (D/d) of transparent circle on Mierocrystalline cellulose-Congo red flat board according to bacterial strain, filtered out 1 plant height temperature bacterium, this bacterial strain is at the transparent circle clearly that occurs in the 2nd day of cultivating, and in cultivation, the 5th day D/d reaches 11.3.This bacterial strain bacterium colony subcircular, surperficial fine hair shape, loose, the edge mycelia is tightr, gray; Base portion brown of light color.
3. utilize the 16SrDNA-PCR technology to carry out molecular biology identification to this bacterial strain, measured the sequence of 16SrDNA, sequence is as shown in SEQ ID No.1, with the sequence homology of the 16SrDNA of the thermophilic carbon monoxide streptomycete of other announcement be 99%, be accredited as thermophilic carbon monoxide streptomycete (Streptomyces thermocarboxydus), called after CF1, and be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on November 23rd, 2012, be called for short CGMCC, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, deposit number is CGMCC No.6873.
Thermophilic carbon monoxide streptomycete CF1 of the present invention can be prepared into microbial inoculum, wherein, and more than the viable count of thermophilic carbon monoxide streptomycete CF1 can be 5,000,000,000/gram, more than being preferably 10,000,000,000/gram.
The present invention also provides the preparation method of described bacteria agent, and it is the thermophilic carbon monoxide streptomycete CF1 of fermentation in liquid fermentation medium, and the fermented liquid obtained is to bacteria agent.
Wherein, preferred fermention medium is No. 1 liquid nutrient medium of Gao Shi.
Preferably, described preparation method can specifically comprise the steps:
By No. 1 culture medium slant of thermophilic carbon monoxide streptomycete CF1 inoculation Zhi Gaoshi, cultivate 48-72h in 50 ℃ of constant temperatures and obtain the inclined-plane seed;
By No. 1 liquid nutrient medium of inclined-plane seed inoculation Zhi Gaoshi, in 50 ℃ of constant temperatures, the 120-200r/min speed oscillation is cultivated 48-72h and is obtained liquid seeds;
By liquid seeds, with 0.5-10%(V/V, every 100ml liquid nutrient medium is inoculated the 0.5-10ml liquid seeds) inoculum size, be inoculated in the productive culture base in liquid fermentation tank, carry out bacterial classification production, in continuing under aeration condition, 50 ℃ of constant temperature culture 3-4 days.After cultivating end, bacterial classification is contained in the container of sterilizing, is placed in shady and cool dry place and preserves.
Preferably, can also the thermophilic carbon monoxide streptomycete CF1 fermented liquid of preparation be carried out to lyophilize according to this area ordinary method, thereby prepare the bacteria agent of lyophilized powder form.
Further, the present invention also provides a kind of composite fungus agent for the animal excrement fermentation, and it comprises subtilis, bacillus megaterium and thermophilic carbon monoxide streptomycete CF1 claimed in claim 1.
Wherein, the viable count of described subtilis is more than 1,000,000,000/gram, and described bacillus megaterium viable count is more than 2,000,000,000/gram, and the viable count of described thermophilic carbon monoxide streptomycete CF1 is more than 2,000,000,000/gram.
Bacillus megaterium of the present invention, subtilis all are commercially available, as purchased from Chinese industrial microbial strains preservation administrative center, bacillus megaterium CICC 20611, subtilis CICC 10090.Certainly the invention is not restricted to this two kinds of bacterial strains.
The new bacterial strain CF1 of inoculation thermophilic carbon monoxide streptomycete of the present invention in the heap body, can improve rapidly heap temperature at northern low temperature season, and can make to pile the high temperature of body maintenance more than 50 ℃ 15 days, shorten fermentation period, can effectively kill pathogen and parasite in animal excrement.After compost fermentation finishes, after compost finishes, nutrient content is high, and the pH8.0 left and right can be used as good plant organic fertilizer and uses.
The composite fungus agent that inoculation comprises the new bacterial strain CF1 of thermophilic carbon monoxide streptomycete of the present invention in the heap body, not only at northern low temperature season, can improve rapidly heap temperature, pliotherm period is long, fermentation period shortens, and compost finishes rear nutrient content loss less, the nutrient content of product significantly improves, and than the product nutrient content of the commercially available EM microbial inoculum of inoculation and Natural compost, significantly increases.
Embodiment
Following examples are used for the present invention is described, but are not used for limiting the scope of the invention.
Unless otherwise noted, " part " described in the present invention refers to " weight part "; Bacillus megaterium bacterial strain CICC 20611, subtilis CICC 10090 bacterial strains are purchased from Chinese industrial microbial strains preservation administrative center.
The preparation of embodiment 1 is thermophilic carbon monoxide drappus microbial inoculum
By No. 1 culture medium slant of thermophilic carbon monoxide streptomycete CF1 inoculation Zhi Gaoshi, cultivate 72h in 50 ℃ of constant temperatures and obtain the inclined-plane seed;
By No. 1 liquid nutrient medium of inclined-plane seed inoculation Zhi Gaoshi, in 50 ℃ of constant temperatures, the 180r/min speed oscillation is cultivated 72h and is obtained liquid seeds;
By liquid seeds with 5%(V/V, i.e. every 100ml liquid nutrient medium inoculation 5ml liquid seeds) inoculum size, be inoculated in the productive culture base in liquid fermentation tank, carry out bacterial classification production, in continuing under aeration condition, 50 ℃ of constant temperature culture 4 days.Fermented liquid after cultivation can be used as microbial inoculum and uses.
No. 1 solid culture based component of described Gao Shi is: in every 1000ml distilled water, add Zulkovsky starch 20.0g, KNO
31.0g, K
2hPO
40.5g, MgSO
47H
2o 0.5g, NaCl 0.5g, FeSO
40.01g, agar 20.0g, adjusting pH is 7.2-7.4.Be distributed in test tube, every pipe 5-10ml, tampon beyond the Great Wall, autoclave sterilization 20min, be put into inclined-plane afterwards.
No. 1 liquid culture based component of described Gao Shi is: in every 1000ml distilled water, add Zulkovsky starch 20.0g, KNO
31.0g, K
2hPO
40.5g, MgSO
47H
2o 0.5g, NaCl 0.5g, FeSO
40.01g regulating pH is 7.0.Be distributed in the 500ml triangular flask every bottle of 150ml, tampon beyond the Great Wall, autoclave sterilization 20min.
Embodiment 2
Fresh cow dung is mixed with the ratio of dry weight 6:1 with crushing straw, or, according to the suitable resize ratio of physico-chemical property of cow dung, sawdust, the C/N that makes mixture is 30, water content is 60%.
The microbial inoculum that embodiment 1 is produced is inoculated in the windrow mixture according to mass ratio 5 ‰, and regularly turning ventilation, carry out the fermented manure fermentation.The heap body that does not access microbial inoculum of the present invention of take is contrast simultaneously.
Inoculating the cow dung of microbiobacterial agent of the present invention compares with the cow dung of Natural compost, heat-up rate is fast, and heap body top temperature is high, and fermentation period is short, its C/N descends rapidly, and these results show to inoculate the degree of becoming thoroughly decomposed that microbiobacterial agent of the present invention can be accelerated cow dung compost in cow dung.
The nutrient content of windrow after the cow dung compost of inoculating the cow dung of microbiobacterial agent of the present invention and Natural compost by comparative analysis finishes, after finding the cow dung compost end of inoculation microbiobacterial agent of the present invention, its full nitrogen, full phosphorus, full potassium and organic content all are significantly higher than the cow dung of Natural compost, and this shows that inoculating microbial inoculum of the present invention in cow dung more is conducive to Nutrient Transformation and preservation in compost.The difference of cow dung on the degree of becoming thoroughly decomposed and each index of nutrient content of inoculating the cow dung of microbiobacterial agent of the present invention and Natural compost is in Table 1 and table 2.
The impact of table 1 microbiobacterial agent of the present invention on the compost maturity degree
Compost initial stage microbial inoculum promotes to decompose free nutritive substance in the heap body, can make heap temperature increase rapidly, and then promote again the breeding of microorganism, temperature rising and microbial reproduction are played to the benign cycle effect, promoting temperature and reach top temperature, when temperature reaches the highest, is that microbial reaction is the fiercest the time, a large amount of organic matters are degraded, and carbon source is converted into CO
2, and cause piling weight, descend.Temperature is the external manifestation of heap body internal-response situation.That temperature rises is faster, high-temperature duration is longer, piles body inside and is fully reacted more fully, more is conducive to the release of the nutrient that is fixed.More abundant at highest temperature elementary reaction, the free nutritive substance and the degradable organic substance that are consumed will be fewer, can not cause fertilizer the seedling phenomenon to occur burning after fertilising.Large amount of organic is after hot stage is degraded, and the after-ripening stage is decomposed in the back to only have a small amount of degradable material.The material that decompose early stage is more much more abundant, just more few more mitigation that after-ripening is decomposed, and temperature will be lower, and outlet temperature can approach room temperature.High-temperature duration is long simultaneously, can kill thoroughly the pathogen in animal excrement.Therefore, the thermophilic carbon monoxide streptomycete CF1 of the present invention's screening is a kind of good high temperature bacterium.
Sampling in time after compost finishes, drill through 40～50cm place sample under the heap surface layer with little earth boring auger, and air-dry rear grinding is measured organic, full nitrogen, full phosphorus, full potassium content, in Table 2.
The nutrient content of table 2 compost sample
Embodiment 3 is for the composite fungus agent of animal excrement fermentation
The subtilis seed liquor 10mL that gets incubated overnight transfers into the 500ml fermention medium, and 28 ℃ ferment 20 hours, and rotating speed is 180rpm.
Consisting of of fermentation of bacillus subtilis substratum: sucrose 10g/L, peptone 5g/L, extractum carnis 3g/L, yeast soaks powder 1g/L, bitter salt 0.5g/L, manganous sulfate 0.0005g/L, pH7.0.
The bacillus megaterium seed liquor 20mL that gets incubated overnight transfers into the 500ml fermention medium, under 30 ℃ of conditions, cultivates 48h.Rotating speed is 180rpm.
The bacillus megaterium fermention medium consists of: extractum carnis 5g/L, peptone 5g/L, sodium-chlor 5g/L, pH7.0.
The fermented liquid for preparing thermophilic carbon monoxide streptomycete CF1 by the method for embodiment 1.
The fermented liquid of the fermented liquid of the fermentation of bacillus subtilis liquid of preparation, bacillus megaterium and thermophilic carbon monoxide streptomycete CF1 is mixed by weight 1:1:2, obtained the composite fungus agent for the animal excrement fermentation.
Fresh cow dung is mixed with the ratio of dry weight 6:1 with sawdust, or, according to the suitable resize ratio of physico-chemical property of cow dung, sawdust, the C/N that makes mixture is 30, water content is 60%.
Inoculate aforesaid composite fungus agent, inoculum size is 0.8%, and, with the commercially available positive contrast of EM microbial inoculum, usings and do not inoculate microbial inoculum as negative control.The results are shown in Table 3 and table 4.
The impact of table 3 complex micro organism fungicide of the present invention on the compost maturity degree
The nutrient content of table 4 compost sample
As shown in Table 3, the composite fungus agent that inoculation comprises thermophilic carbon monoxide streptomycete CF1 of the present invention, the heap body can complete fermentation with interior at 23 days, shifted to an earlier date 10-12 days than the commercially available EM microbial inoculum of inoculation and Natural compost and completed fermentation, and the high temperature extended period is long, the degree of becoming thoroughly decomposed after the compost end is good.As shown in Table 4, the composite fungus agent that inoculation comprises thermophilic carbon monoxide streptomycete CF1 of the present invention, in the rear windrow of compost end, organic content, full nitrogen, full phosphorus and full potassium content are all higher than the windrow of inoculating commercially available EM microbial inoculum and Natural compost.Its organic content of composting production of the composite fungus agent that wherein inoculation comprises thermophilic carbon monoxide streptomycete CF1 of the present invention and total nutrient content are all higher than the regulation in national standard (organic fertilizer NY-525-2012), and inoculate the composting production of commercially available EM microbial inoculum, just be up to state standards (organic fertilizer NY-525-2012), the product of Natural compost be not up to state standards (organic fertilizer NY-525-2012).
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the technology of the present invention principle; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (7)
1. the new bacterial strain CF1 of a thermophilic carbon monoxide streptomycete (Streptomyces thermocarboxydus), its preserving number is CGMCC No.6873.
2. the bacteria agent that contains the described bacterial strain of claim 1.
3. bacteria agent according to claim 2, wherein the viable count of thermophilic carbon monoxide streptomycete CF1 is more than 5,000,000,000/gram.
4. the method for preparing claim 2 or 3 described bacteria agents, it is the thermophilic carbon monoxide streptomycete CF1 of fermentation in liquid fermentation medium, and the fermented liquid obtained is to bacteria agent.
5. method as claimed in claim 4, it also comprises thermophilic carbon monoxide streptomycete CF1 fermented liquid is carried out to lyophilize, the lyophilized powder be prepared into is bacteria agent.
6. the composite fungus agent for animal excrement fermentations, it comprises subtilis, bacillus megaterium and thermophilic carbon monoxide streptomycete CF1 claimed in claim 1.
7. microbial inoculum as claimed in claim 6, it is characterized in that, the viable count of described subtilis is more than 1,000,000,000/gram, and described bacillus megaterium viable count is more than 2,000,000,000/gram, and the viable count of described thermophilic carbon monoxide streptomycete CF1 is more than 2,000,000,000/gram.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104531557A (en) * | 2014-12-04 | 2015-04-22 | 江苏省血吸虫病防治研究所 | Streptomyces subrutilus capable of generating molluscicide active substance and application thereof |
| CN104988095A (en) * | 2015-07-03 | 2015-10-21 | 北京农学院 | Thermophilic carbon monoxide streptomycete low-temperature subspecies Dstr3-3 and application thereof |
| CN105567607A (en) * | 2016-03-03 | 2016-05-11 | 北京林业大学 | High-temperature-resistant garden waste decomposing bacteria ST3 and application thereof |
| CN105567612A (en) * | 2016-03-05 | 2016-05-11 | 北京林业大学 | Preparation and application of garden waste compound degrading microbial agent |
| CN106007847A (en) * | 2016-05-17 | 2016-10-12 | 许祚员 | Efficient bio-organic fertilizer for pomegranate trees and preparation method of fertilizer |
| CN109136156A (en) * | 2018-10-10 | 2019-01-04 | 皖西学院 | Solid composite bacteria agent and preparation method thereof and the application in officinal dendrobium stem plantation |
| CN109517754A (en) * | 2018-11-20 | 2019-03-26 | 上海交通大学 | A method of high temperature bacterial strain is isolated and purified using common biochemical equipment |
| CN111378604A (en) * | 2020-03-17 | 2020-07-07 | 山东中农普宁药业有限公司 | Thermoactinomyces hopeiensis and application thereof |
| CN114196583A (en) * | 2021-12-15 | 2022-03-18 | 天津科技大学 | Thermophilic carbon monoxide chain mould, synergistic biological agent containing same and application |
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| CN104531557A (en) * | 2014-12-04 | 2015-04-22 | 江苏省血吸虫病防治研究所 | Streptomyces subrutilus capable of generating molluscicide active substance and application thereof |
| CN104988095B (en) * | 2015-07-03 | 2019-01-22 | 北京农学院 | One plant of thermophilic carbon monoxide streptomycete low temperature subspecies Dstr3-3 and its application |
| CN104988095A (en) * | 2015-07-03 | 2015-10-21 | 北京农学院 | Thermophilic carbon monoxide streptomycete low-temperature subspecies Dstr3-3 and application thereof |
| CN105567607B (en) * | 2016-03-03 | 2019-11-01 | 北京林业大学 | One plant of high temperature resistant garden waste decomposer ST3 and its application |
| CN105567607A (en) * | 2016-03-03 | 2016-05-11 | 北京林业大学 | High-temperature-resistant garden waste decomposing bacteria ST3 and application thereof |
| CN105567612A (en) * | 2016-03-05 | 2016-05-11 | 北京林业大学 | Preparation and application of garden waste compound degrading microbial agent |
| CN105567612B (en) * | 2016-03-05 | 2019-08-02 | 北京林业大学 | A kind of degradation composite bacteria agent preparation of garden waste and application |
| CN106007847A (en) * | 2016-05-17 | 2016-10-12 | 许祚员 | Efficient bio-organic fertilizer for pomegranate trees and preparation method of fertilizer |
| CN109136156A (en) * | 2018-10-10 | 2019-01-04 | 皖西学院 | Solid composite bacteria agent and preparation method thereof and the application in officinal dendrobium stem plantation |
| CN109517754A (en) * | 2018-11-20 | 2019-03-26 | 上海交通大学 | A method of high temperature bacterial strain is isolated and purified using common biochemical equipment |
| CN111378604A (en) * | 2020-03-17 | 2020-07-07 | 山东中农普宁药业有限公司 | Thermoactinomyces hopeiensis and application thereof |
| CN111378604B (en) * | 2020-03-17 | 2022-06-10 | 山东中农普宁药业有限公司 | Thermoactinomyces hopeiensis and application thereof |
| CN114196583A (en) * | 2021-12-15 | 2022-03-18 | 天津科技大学 | Thermophilic carbon monoxide chain mould, synergistic biological agent containing same and application |
| CN114196583B (en) * | 2021-12-15 | 2023-11-17 | 天津科技大学 | Streptomyces thermophilus, synergistic biological agent containing same and application thereof |
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