CN110923175B - Bacillus and application thereof in reduction and resource production of organic fertilizer by kitchen waste - Google Patents
Bacillus and application thereof in reduction and resource production of organic fertilizer by kitchen waste Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
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- C12R2001/10—Bacillus licheniformis
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- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F5/00—Fertilisers from distillery wastes, molasses, vinasses, sugar plant or similar wastes or residues, e.g. from waste originating from industrial processing of raw material of agricultural origin or derived products thereof
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- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
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Abstract
The invention discloses bacillus and application of bacillus in reduction and recycling of kitchen waste to produce organic fertilizer, and belongs to the technical field of biological bacterial fertilizer. According to the invention, a bacillus with obvious antagonistic action on fusarium is obtained by screening and is used for composting kitchen waste, so that the kitchen waste can be rapidly reduced and recycled. The bacterial manure prepared by the method can improve the physical, chemical and biological characteristics of soil, inhibit plant diseases and have important industrial application prospects.
Description
Technical Field
The invention relates to bacillus and application of bacillus in reduction and recycling of kitchen waste to produce organic fertilizer, and belongs to the technical field of biological bacterial fertilizer.
Background
The kitchen waste is rich and comprehensive in nutrition, high in water content, easy to decay, capable of generating pungent smell, capable of causing bad influence on environmental sanitation, and easy to grow various pathogenic microorganisms and pests such as mosquitoes and the like. Due to the Chinese eating habits. The table is extremely wasteful, and generates a huge amount of kitchen waste every day. Great environmental pollution is caused, and a rapid and efficient reduction treatment method for the kitchen waste is urgently needed.
In agricultural production in China, the organic fertilizer has huge gaps, comprehensive nutrients and high organic matter content, can improve soil organic matters, improve soil granular structure, promote the propagation of soil microorganisms, loosen and ventilate the soil, enhance the fertilizer retention performance and buffer capacity of the soil, enhance the water retention, fertilizer retention and fertilizer supply capacity, and can further germinate seeds, grow roots and absorb nutrients, thereby improving the quality of agricultural products.
Bacillus attracts great attention as a bacterium with a prospect of biocontrol application. The bacillus inhibits the growth of pathogenic bacteria through competition of nutrient and space sites. The bacteriocin produced by the bacillus has certain inhibition effect on mould, yeast and bacteria in pathogenic bacteria. Chitinase secreted by the bacillus has obvious biocontrol effect on decomposing the cell walls of pathogenic bacteria, and antibacterial protein is secreted to inhibit the production and germination of spores of the pathogenic bacteria, so that hyphae are deformed, the cell walls are dissolved, and protoplasm is leaked. The bacillus generates signal protein related to the expression of plant resistance protein synthetic gene, and has the functions of promoting plant growth and inducing plant resistance.
The kitchen waste is rich in nutrition and has the possibility of producing organic fertilizer through fermentation.
According to the invention, a bacillus JSDD3012 capable of efficiently fermenting kitchen waste to produce an organic fertilizer is obtained through screening, a process method for producing the organic fertilizer by JSDD3012 fermenting kitchen waste is developed, the reduction and the resource utilization of the kitchen waste are realized, and the produced organic fertilizer meets the national organic fertilizer standard NY525-2012 and the biological organic fertilizer standard NY 884-2012. And the content of bacillus in per gram of dry matter in the produced organic fertilizer exceeds 109And (4) respectively.
Disclosure of Invention
The invention aims to protect a Bacillus for efficiently fermenting kitchen waste to produce organic fertilizer, which is classified and named as Bacillus licheniformis and is preserved in Guangdong province microorganism strain preservation center in 2019, 7 and 11 days, wherein the preservation number is GDMCC No: 60726, the preservation address is No. 59 building 5 of No. 100 Dazhong Jie-Lu-100 Guangzhou city.
It is a second object of the present invention to provide a microbial preparation containing said bacillus.
In one embodiment, the microbial preparation comprises said bacillus in a cellular or spore form.
The third purpose of the invention is to provide a process method for producing an organic fertilizer by fermenting kitchen waste with bacillus
The fourth purpose of the invention is to provide an organic fertilizer containing a large amount of the bacillus.
In one embodiment, the fertilizer is prepared by inoculating the bacillus to kitchen waste serving as a raw material and composting.
The fifth purpose of the invention is to provide a treatment method of kitchen waste, which takes the bacillus as a fermentation microorganism and performs solid state fermentation for 20-72 h at 25-30 ℃.
In one embodiment of the invention, the fermentation is a staged fermentation comprising a first stage and a second stage;
the first stage is to mealMixing kitchen waste and bran at a mass ratio of 2:1, inoculating with the strain concentration of (1-8) x 106Heating 3kg of CFU/g of bacillus fermenting substrate to 80 ℃ within 30 minutes, keeping for 8-10 minutes, after aeration cooling for 20-30 minutes, heating to 80 ℃ again within 30 minutes, keeping for 8-10 minutes, then aeration cooling, keeping for 28-30 ℃, continuing the fermentation process for 70-75 hours, and stirring for 120 turns per hour;
and (3) discharging bacterial manure which accounts for not more than 15% of the total mass of the fermentation system after the first-stage fermentation in the second stage, adding crushed kitchen waste with the same mass as that of the first-stage fermentation, heating to 80 ℃ within 30 minutes, keeping for 10 minutes, cooling by ventilation, repeating heating to 80 ℃ after 30 minutes, keeping for 10 minutes, cooling by ventilation, keeping for 28-30 ℃, continuing the fermentation process for 22 hours, and stirring for 120 turns per hour in the tank.
In one embodiment of the invention, the fermented bacterial manure is air-dried to a moisture content of less than 10%.
In one embodiment of the invention, the fermentation is an inoculation concentration greater than 1X 109CFU/ml seed solution.
In one embodiment of the present invention, the seed liquid is prepared as follows: a single colony of Bacillus was inoculated into LB liquid medium and shake-cultured at 28 ℃ and 140rpm for 2 days.
In one embodiment of the invention, the fermentation is performed by taking kitchen waste and bran in a mass ratio of (2-3): 1 as raw materials and adjusting the water content to 45-50%.
In one embodiment of the present invention, the kitchen waste is pulverized to 20 mesh.
In one embodiment of the invention, the method repeats the second stage fermentation process.
The invention also claims application of the bacterial manure prepared by the method in crop cultivation.
Has the advantages that: the invention has the following advantages:
1. the invention obtains a bacillus for producing organic fertilizer by efficiently fermenting kitchen waste through screening;
2. the strain and the process of the invention enable the dehydration rate of the kitchen waste to reach 85-90%. The high-efficiency reduction of the kitchen waste is realized.
3. The organic fertilizer produced by the method reaches the national organic fertilizer standard NY525-2012 and the biological organic fertilizer standard NY884-2012, and the rapid recycling of the kitchen waste is realized.
4. The fermentation process method is easy to operate, simple in equipment and suitable for industrial application.
5. The content of bacillus in the organic fertilizer prepared by the invention exceeds 109CFU/gram.
Biological material preservation
A strain of Bacillus licheniformis (Bacillus licheniformis) is classified and named as Bacillus licheniformis and has been deposited in Guangdong province microorganism culture collection center in 7-11.2019, and the deposit number is GDMCC No: 60726, the preservation address is No. 59 building 5 of No. 100 Dazhong Jie-Lu-100 Guangzhou city.
Detailed Description
The determination method comprises the following steps:
protein content: kjeldahl method (GB/T5009.5-2003);
fat content: acid hydrolysis methods (GB/T5009.6-2003);
organic matter content: measuring the content of organic matters by adopting a burning method;
water content: measuring by adopting a weight loss method;
the degradation rate is defined as: in the degradable component, the mass of the degraded portion is a percentage of the initial mass of the degradable component.
The formula of the culture medium is as follows:
LB solid medium: 5.0g of peptone, 3.0g of beef extract, 5.0g of sodium chloride, 1000mL of distilled water and 20g of agar.
LB liquid medium: 5.0g of peptone, 3.0g of beef extract, 5.0g of sodium chloride and 1000mL of distilled water.
Example 1 screening and identification of strains
(1) 10g of collected soil sample near a canteen of an enterprise is put into a triangular flask with 90ml of sterile water containing glass beads, is subjected to shaking culture at the temperature of 30 ℃ for 30min at the speed of 200r/min, is subjected to water bath at the temperature of 80 ℃ for 10min, is diluted step by step according to gradient, and is coated on an LB flat plate. Culturing the coated LB plate at 28 deg.C for 3 days, picking different single colonies, repeating streaking separation and purification on LB plate for 3 times, and performing microscopic examination to select strains with spores.
(2) Respectively inoculating the bacillus separated in the step (1) into LB culture media, and culturing for 16h to obtain seed solutions;
(3) and (3) inoculating 10% of the seed liquid obtained in the step (2) by taking the kitchen waste collected in the canteen of the enterprise as a culture medium, fermenting for 48h at 55 ℃, detecting the fat content change of a fermentation system, and screening 16 bacillus strains with relatively excellent degradation effect in total, wherein the bacillus strains are shown in table 1:
TABLE 1 degradation rate of fat by Bacillus
(3) And selecting strains with the kitchen waste degradation rate of more than 70% for 16S rDNA identification, wherein the serial numbers 3 and 8 are Bacillus licheniformis, the serial number 4 is Bacillus amyloliquefaciens, and the serial number 12 is Bacillus pumilus.
(4) Inoculating Bacillus with biological control function into 100mL LB liquid medium, shake culturing at 37 deg.C and 140rpm for 2 days, subpackaging for two days, placing in 25% glycerol, and preserving at-80 deg.C for a long time.
Example 2
(1) Salmonella typhimurium SL1344 is subjected to activation culture in LB culture medium and then is subjected to shake culture at 37 ℃ and 225rpm for 12 hours.
(2) After the activation and subculture of 4 strains of Bacillus selected in example 1, the cells were cultured at 37 ℃ in LB medium for 14 hours and centrifuged, and the supernatant of the culture was collected and used as a control group in LB medium without inoculation. The culture supernatant and the control group were filtered through a sterile filter with a pore size of 0.22 μm.
(3) To 5mL of each of the culture supernatants of the above 4 strains of Bacillus, 50. mu.L of a concentrate of 1X 10 cells was added9CFU/mL Salmonella SL1344 bacterial solution, standing and culturing at 37 ℃ for 24h, and determining OD of co-culture600。
TABLE 2 growth inhibitory Effect of Bacillus on Salmonella
The result shows that the bacillus licheniformis 3 has good inhibiting effect on salmonella, and the bacillus licheniformis is preserved by patented strains.
Example 3 application of biocontrol strains to kitchen waste composting
(1) The bacillus licheniformis with biological control function obtained by screening in the example 2 is inoculated into 1L LB liquid medium and cultured for 2 days in a shaking table at the temperature of 28 ℃ and the rpm of 140 to obtain the bacterial concentration of 109CFU/mL of the broth was used as the seed broth for the next fermentation.
(2) 300 kg of kitchen waste is taken, crushed, sieved by a 20-mesh sieve and mixed with about 150 kg of bran, and the water content of the mixture is 45-50%.
(3) And (3) putting the mixture of the kitchen waste and the bran obtained in the step (2) into a solid-state fermentation tank developed by Jiangsu Daodao Bioenvironmental technology Limited company, and inoculating 1 kg of the bacillus licheniformis liquid obtained in the step (1) into one hundred kg of the kitchen waste.
(4) Heating the mixture of the kitchen waste, the bran and the strains to 80 ℃ in a fermentation tank, keeping for 10 minutes, ventilating and cooling, repeatedly heating to 80 ℃ after half an hour, keeping for 10 minutes, then ventilating and cooling, keeping for 28-30 ℃, stirring for 120 times per hour in the tank, and continuing the whole fermentation process for 72 hours.
(5) And after fermentation culture is carried out for 72h, ventilation drying is carried out in the fermentation tank for 2h, so that the water content of the bacillus bacterial manure is lower than 10%, and the organic fertilizer is obtained.
(6) Discharging organic fertilizer accounting for 10-15% of the total amount of the kitchen waste, mixing the remaining organic fertilizer rich in bacillus serving as a strain with the crushed kitchen waste, heating to 80 ℃ in a fermentation tank, keeping for 10 minutes, cooling by ventilation, heating to 80 ℃ again after 30 minutes, keeping for 10 minutes, cooling by ventilation, keeping for 28-30 ℃, fermenting for 22 hours, and stirring for 120 times per hour in the tank. And after fermentation is finished, the tank is ventilated and dried for 2 hours, so that the water content of the organic fertilizer is lower than 10%.
(7) And (5) repeating the steps (5) to (6), realizing daily feeding of the kitchen waste and daily discharge of the organic fertilizer, and realizing continuous treatment of the kitchen waste and continuous output of the organic fertilizer.
Particularly, the first culture of the bacillus needs 72 hours, the rest needs only 22 hours, a large amount of mixed bran is needed for the first time, the rest does not need to be mixed with a large amount of bran, and the proper amount of supplement is carried out according to the condition of the kitchen waste only when the water content can not reach the requirement (40-50%).
The organic fertilizer prepared in the embodiment is detected, the detection method refers to NY525-2012 standards and NY884-2012 standards, and the result shows that the content of bacillus in each g of organic fertilizer exceeds 1 × 109A CFU; after fermentation for 22 hours, the protein degradation rate is 62.3 percent, the fat degradation rate is 65.37 percent, organic matters (protein, fat and total sugar) are 67.5 percent, total nutrients are 12.8 percent, water is 7.3 percent, the number of faecal coliform bacteria in the fertilizer is 3 per gram, and the death rate of ascarid eggs is 99.99 percent. The organic fertilizer meets the national organic fertilizer standard NY525-2012 and the biological organic fertilizer standard NY 884-2012.
Comparative example 1:
composting was carried out as in example 2, with the difference that the water content was adjusted to 30%, and the results showed that the Bacillus content per g of organic fertilizer was 1X 106A CFU; the protein degradation rate is 22.8%, the fat degradation rate is 16.59%, the organic matter content is 32.45%, the total nutrient content is 5.3%, and the water content is 9.2%, which does not meet the national bio-organic fertilizer NY884-2012x standard.
Comparative example 2
Composting was carried out as in example 2, with the difference that the fermentation time in the first stage was reduced to 24h, and the results showed a Bacillus content of 1X 10 per g of organic fertilizer7A CFU; the protein degradation rate is 31.5%, the fat degradation rate is 25.07%, the organic matter content is 37.45%, the total nutrient content is 7.1%, the water content is 8.3%, and the standard of the national bio-organic fertilizer NY884-2012x is not met。
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (7)
1. A treatment method of kitchen waste is characterized in that bacillus is used as fermentation microorganism and inoculated to a substrate containing the kitchen waste for solid state fermentation;
the Bacillus is classified and named as Bacillus licheniformis, is preserved in Guangdong province microorganism strain preservation center in 2019, 7 and 11 days, and the preservation number is GDMCC No: 60726, the preservation address is No. 59 building 5 of the Fuzhou first Liehuo No. 100 yard;
the fermentation is a staged fermentation comprising a first stage and a second stage;
in the first stage, the kitchen waste and the bran are mixed by the following steps of (2-3): 1, the inoculation concentration is (1-8) multiplied by 106Heating the bacillus of the CFU/g fermentation substrate to 70-80 ℃, keeping for 8-10 minutes, cooling for 20-30 minutes in an aeration manner, then repeatedly heating to 70-80 ℃, keeping for 8-10 minutes, cooling in an aeration manner, and fermenting for 70-75 hours at 28-30 ℃;
and (3) discharging bacterial manure which accounts for not more than 15% of the total mass of the fermentation system after the first-stage fermentation in the second stage, adding the crushed kitchen waste with the same mass as that of the first-stage fermentation, heating to 70-80 ℃, keeping for 5-15 minutes, cooling by ventilation, repeatedly heating to 70-80 ℃, keeping for 5-15 minutes, cooling by ventilation, and continuously fermenting for 22 hours at 28-30 ℃.
2. The method for treating kitchen waste according to claim 1, characterized in that the second stage is repeated several times.
3. The method for treating the kitchen waste according to claim 1, wherein the water content of the mixture of the kitchen waste and the bran is adjusted to 45-50% before inoculation.
4. The method for treating the kitchen waste according to claim 1, characterized in that fermented bacterial manure is dried by ventilation until the moisture content is lower than 10%.
5. A microbial preparation comprising the Bacillus of claim 1.
6. The use of the bacillus of claim 1, or the microbial preparation of claim 5, in the fermentation of kitchen waste to produce organic fertilizer.
7. An organic fertilizer containing a large amount of the bacillus of claim 1, which is prepared by the method of any one of claims 1 to 4.
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Citations (7)
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| CN103820350A (en) * | 2012-11-16 | 2014-05-28 | 中国科学院生态环境研究中心 | Method for producing bacillus amyloliquefaciens microbial fertilizer through food waste recycling |
| CN105732143A (en) * | 2016-03-09 | 2016-07-06 | 广东海洋大学 | Method for producing organic fertilizer without pile turning |
| CN106190900A (en) * | 2016-07-15 | 2016-12-07 | 标优美生态工程股份有限公司 | A kind of Bacillus licheniformis and the application in changing food waste thereof |
| CN108004169A (en) * | 2017-12-18 | 2018-05-08 | 广州王道生物科技有限公司 | Bacillus licheniformis ZL-1 and its application |
| CN109182215A (en) * | 2018-10-17 | 2019-01-11 | 湖南省林业科学院 | One bacillus licheniformis bacterial strain and its application in inhibition bacterial pathogen |
| CN110218680A (en) * | 2019-06-24 | 2019-09-10 | 北京瑞斯壮环保设备有限公司 | Composite bacteria agent and its method for handling kitchen garbage |
| CN110343631A (en) * | 2019-05-06 | 2019-10-18 | 南开大学 | A kind of kitchen garbage bio-fermentation agents and application |
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Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103820350A (en) * | 2012-11-16 | 2014-05-28 | 中国科学院生态环境研究中心 | Method for producing bacillus amyloliquefaciens microbial fertilizer through food waste recycling |
| CN105732143A (en) * | 2016-03-09 | 2016-07-06 | 广东海洋大学 | Method for producing organic fertilizer without pile turning |
| CN106190900A (en) * | 2016-07-15 | 2016-12-07 | 标优美生态工程股份有限公司 | A kind of Bacillus licheniformis and the application in changing food waste thereof |
| CN108004169A (en) * | 2017-12-18 | 2018-05-08 | 广州王道生物科技有限公司 | Bacillus licheniformis ZL-1 and its application |
| CN109182215A (en) * | 2018-10-17 | 2019-01-11 | 湖南省林业科学院 | One bacillus licheniformis bacterial strain and its application in inhibition bacterial pathogen |
| CN110343631A (en) * | 2019-05-06 | 2019-10-18 | 南开大学 | A kind of kitchen garbage bio-fermentation agents and application |
| CN110218680A (en) * | 2019-06-24 | 2019-09-10 | 北京瑞斯壮环保设备有限公司 | Composite bacteria agent and its method for handling kitchen garbage |
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