CN114958681A - Composite microbial inoculum for decomposing waste animal carcasses, application of composite microbial inoculum and method for decomposing waste animal carcasses through high-temperature aerobic fermentation - Google Patents
Composite microbial inoculum for decomposing waste animal carcasses, application of composite microbial inoculum and method for decomposing waste animal carcasses through high-temperature aerobic fermentation Download PDFInfo
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Abstract
The invention belongs to the field of waste animal carcass treatment, and discloses a composite microbial inoculum for decomposing waste animal carcasses, application of the composite microbial inoculum and a method for decomposing the waste animal carcasses through high-temperature aerobic fermentation. According to the invention, bacillus stearothermophilus, bacillus amyloliquefaciens, bacillus licheniformis, bacillus calpithecus, geobacillus stearothermophilus, Thermotoga maritima, tennis thermophilus and archaebacterium thermophilus are mixed according to a specific proportion to obtain a composite microbial inoculum capable of decomposing waste animal carcasses. The results of the embodiment show that the composite microbial inoculum provided by the invention can form a bacterial biofilm after being inoculated on an adsorption medium, and the bacterial biofilm can be mixed with the waste animal carcasses to rapidly degrade the waste animal carcasses at a high temperature of 70-80 ℃, so that the problems of reduction and recycling of the waste animal carcasses are solved, and the aim of rapidly reducing and treating the animal carcasses is fulfilled.
Description
The application is a divisional application with application date of 2014, 08 and 06, application number of 201410384531.5, and invention name of 'a method for decomposing waste animal carcasses by high-temperature aerobic fermentation'.
Technical Field
The invention belongs to the field of waste animal carcass treatment, and particularly relates to a composite microbial inoculum for decomposing waste animal carcasses, application of the composite microbial inoculum and a method for decomposing the waste animal carcasses through high-temperature aerobic fermentation.
Background
With the rapid development of livestock and poultry breeding, slaughtering and meat processing enterprises, the waste animal carcasses produced have placed a great deal of pressure on environmental protection. According to the stipulations of animal epidemic prevention Law of the people's republic of China: at present, the methods for harmless, quantitative-reduction, resource-recycling and harmless treatment of the waste animal carcasses comprise sanitary landfill, composting and incineration. From the technical process, the methods pursue the same target, but have advantages and disadvantages, and have larger difference of technical difficulty degree:
(1) a burning method; the method can generate secondary pollution and high-toxicity tri-cause (carcinogenic, teratogenic and mutagenic) gas which are harmful to human health.
(2) High temperature and high pressure method; the method is characterized in that high-temperature and high-pressure equipment is used for treating animal carcasses, the equipment belongs to special equipment, a boiler worker is specially equipped, waste water and waste gas generated by the equipment can reach the environmental protection standard only by being reprocessed by the special equipment, and the product of the method needs solid-liquid separation and needs to be added with separation equipment.
(3) Burying in a sanitary way; the method has large floor area and permanently occupies land, bubbles and liquid leakage are caused by the putrefaction gas generation of the dead body during landfill, and great potential safety hazards exist on underground water due to chemicals (quicklime and bleaching powder) used during landfill.
(4) According to the traditional fermentation method, the mixture of rice chaff, sawdust, straw, corncob and the like is made into compost for fermentation, the compost can be fermented generally for 1-3 months, the time period is long, land resources are occupied for a long time, the fermentation process generates odor pollution, the fermentation product is an organic fertilizer, and the economic benefit of the organic fertilizer is poor.
The technology for treating the waste animal carcasses of the thermophilic bacteria is an organic waste treatment technology with good development prospect at present. The technology for treating the waste animal carcasses by using high-temperature bacteria is a method for biologically degrading the waste animal carcasses into animal feed raw materials, namely meat and bone meal by using microorganisms or artificial inoculants such as bacteria, bacillus, Thermotoga maritima, archaea and the like widely distributed in the nature under the condition of artificial control and ensuring certain water content, oxygen supply, temperature and the like. The main treatment measures and ways are as follows: (1) because of the implementation of industrial large-scale cultivation, slaughtering and processing, the consumption of animal carcasses such as livestock and poultry is large, the method has the advantages of high reaction speed, high treatment efficiency and less land resource occupation; (2) the waste animal carcasses are prepared into high-quality animal feed raw material meat and bone powder, so that energy and nutrients in the meat and bone powder are recycled, and the resource level is high; (3) the animal feed raw material meat and bone meal prepared by fermenting the waste animal carcasses can prevent the waste animals from being sold in the market as fresh animal meat from the source, and resources can be recycled while food safety is guaranteed. (4) The high-temperature bacteria waste animal treatment technology not only solves the treatment of waste animals, but also kills pathogenic microorganisms and viruses after the generated animal feed raw materials are continuously fermented at high temperature for a long time, reduces the pollution of bacterial metabolites to products, and ensures the use safety of resource products (see appendix 2, salmonella analysis report of feed quality supervision, inspection and test center (Guangzhou) of the department of agriculture) (see appendix 3, blue ear disease analysis and test report of diagnosis center of animal health research institute of Guangdong province). (5) The waste animals are completely decomposed into amino acids which are easy to digest and absorb by the animals after high-temperature fermentation, the sum of the hydrolyzed amino acids exceeds 40% (see appendix 1, the hydrolyzed amino acid analysis report of the Guangzhou analysis test center in China), and the feed prepared from the fermentation product has the characteristics of strong palatability and good taste. The ability of microorganisms to break down waste animal carcasses is surprising. It can be said that all the waste animals existing in the natural world are almost decomposed by microorganisms. Some bacterial species, such as Thermotoga maritima (Thermotoga spp.) degrade fats, proteins and cellulose and are metabolized using any of these organic substances as the sole carbon source. The application of the high-temperature bacteria in the reduction, recycling and harmless treatment of the waste animal carcasses is mainly the difficult problems of screening, separation, optimization, combined culture, biological membrane immobilization and the like.
Disclosure of Invention
The invention aims to provide a composite microbial inoculum for decomposing waste animal carcasses, application thereof and a method for decomposing the waste animal carcasses by high-temperature aerobic fermentation, so that the purposes of effectively reducing, recycling and harmlessness are achieved, and the waste animal carcasses can be decomposed to produce excellent animal feed raw material meat and bone meal.
The method for decomposing the waste animal carcasses through high-temperature aerobic fermentation is characterized by comprising the following steps of:
inoculating bacillus stearothermophilus, bacillus amyloliquefaciens, bacillus licheniformis, bacillus calpitis, geobacillus stearothermophilus, Thermotoga maritima, Thermococcus thermophilus and archaebacteria thermophilus to an adsorption medium, uniformly stirring, culturing until the adsorption medium is changed from yellow to brown, mixing the adsorption medium with waste animal carcasses, and fermenting to obtain the animal feed raw material meat and bone meal, wherein the adsorption medium is a mixture consisting of perlite particles and rice bran.
The method for decomposing the waste animal carcasses by high-temperature aerobic fermentation of the present invention is preferably: fermenting and culturing bacillus stearothermophilus, bacillus amyloliquefaciens, bacillus licheniformis, bacillus calpitis, geobacillus stearothermophilus, geobacillus marinus, Thermotoga maritima, Anemophilus thermophilus and archaebacterium thermophilus in a stable period by using a fermentation culture medium according to the mass ratio of 1: 1: 1: 1: 1: 2: 2: 2 to form a composite microbial inoculum, and mixing the composite microbial inoculum and an adsorption medium according to a mass ratio of 1: 2, uniformly mixing, culturing under the conditions of 70-80 ℃ and 20% of relative humidity until the adsorption medium is changed from yellow to brown, mixing the adsorption medium containing the composite microbial inoculum after color change with the waste animal carcasses, adding and mixing for multiple times according to the treatment capacity of 100-300 kg of waste animal carcasses per day of the adsorption medium containing the composite microbial inoculum, and fermenting at the temperature of 70-80 ℃ until fermentation is finished, wherein the fermented product is the animal feed raw material meat and bone meal;
the fermentation medium contains 5g of beef extract powder, 10g of peptone, 5g of yeast extract, 5g of sodium chloride and the balance of water per liter, and the pH value is 7.0. Sterilizing and disinfecting for later use.
The adsorption medium is prepared from perlite particles and rice bran according to a mass ratio of 1: 1, mixing the mixture.
Bacillus stearothermophilus (Bacillus stearothermophilus) is milky white, rod-shaped, gram-positive, sporulated and optimal in growth temperature of 70-80 ℃. The strain belongs to the strains in the prior art, and can be purchased from biological companies or depository institutions, such as Bacillus stearothermophilus CICICIM B0553.
Bacillus amyloliquefaciens (Bacillus amyloliquefaciens) is milky, rod-shaped, gram-positive, sporulated and optimal in growth temperature of 70 ℃. The strain belongs to a strain in the prior art, and can be purchased from biological companies or depository institutions, such as Bacillus amyloliquefaciens CICIM B4311 of the industrial microbial data platform of colleges and universities in China.
Bacillus licheniformis (Bacillus licheniformis) is milk white, rod-shaped, gram-positive, spore-forming, and optimal growth temperature is 70 ℃. The strain belongs to the strain in the prior art, can be purchased from biological companies or depository institutions, such as Bacillus licheniformis CICICIM B1455 of industrial microbial data platform of colleges and universities in China.
Bacillus thermoacidophilus (Bacillus thermoacidophilus) is milk white, rod-shaped, gram-positive, has spores and has the optimum growth temperature of 70-75 ℃. The strain belongs to the strain in the prior art, can be purchased from biological companies or depository institutions, such as Bacillus thermoleovarans CICICIM B0614 of the industrial microbial data platform of colleges and universities in China.
Geobacillus stearothermophilus (Geobacillus spp.) is milk white, rod-shaped, gram-positive, has spores and has the optimal growth temperature of 70 ℃. The strain belongs to the strains in the prior art, can be purchased from biological companies or depository institutions, such as Geobacillus stearothermophilus with the preservation number GIM1.543 of Guangdong provincial microorganism culture depository.
Thermotoga maritima (Thermotogasp.), yellow white, round, has a special sheath structure and the optimal growth temperature is 80 ℃. The strain belongs to the strains in the prior art (such as the papers: clone and expression of Thermotoga maritima xynB gene, purification of recombinant xylanase and the enzymological properties thereof, Jiangzheng, Chinese agriculture university, published in 2001), can be purchased from biological companies or depositories, and is also held by the applicant and guaranteed to be provided to the public within 20 years from the application date.
Pyrococcus thermophilus (Dictyoglycomus thermophilum) is milky and filamentous, and the optimum growth temperature is 90 deg.C. The strain belongs to the strains in the prior art, is disclosed in various documents, can be purchased from biological companies or depositories, is also held by the applicant and is guaranteed to be provided to the public within 20 years from the filing date.
Thermophilic archaea (Pyrococcusfuriosus) is milky white, rod-shaped, and has an optimal growth temperature of 80 ℃. The strain belongs to a strain in the prior art (for example, a paper is disclosed in 76-81 in the journal of the parent expression and the enzymatic property analysis of the lactase gene of thermophilic archaea Pyrococcus furiosus, the parent of the Setarian Juanjuan lovely plum Piffir of Seisakura, China agricultural science and technology guide, 2008) and can be purchased from a biological company or a preservation organization, and the strain is also held by the applicant and is guaranteed to be provided to the public within 20 years from the application date.
(1) The genetic stability test shows that the 8 high-temperature aerobic fermentation bacteria can keep stable within 15 generations in genetics.
(2) Antibiotic susceptibility determination the antibiotics to which the 8 high temperature aerobic fermentation bacteria are susceptible are chloramphenicol, tetracycline, penicillin G, cephalothin (cepham I), gentamicin, ofloxacin (ofloxacin), polymyxin B, clindamycin (clindamycin), ketoconazole, miconazole/dackerine and clotrimazole.
(3) For the measurement of the acute skin irritation of the rabbits, the acute skin irritation of the rabbits is slightly irritative by 8 high-temperature aerobic fermentation bacteria.
(4) The determination result of the acute oral toxicity of the mice is more than 2000mg/kg by 8 high-temperature aerobic fermentation bacteria. The above results all indicate that the 8 kinds of high temperature aerobic fermentation bacteria of the present invention are suitable for the degradation treatment of waste animal carcasses, and the end product (fermented animal feed raw material meat and bone meal) obtained by the degradation treatment does not adversely affect the living environment of human and animals.
The invention preferably selects 8 bacteria, the bacteria are mixed according to a certain proportion to form a composite microbial inoculum, the composite microbial inoculum forms a bacterial biofilm on an adsorption medium, and the bacterial biofilm can rapidly degrade the waste animal carcasses at the high temperature of 70-80 ℃ after being mixed with the waste animal carcasses, thereby solving the problems of reduction and resource of the waste animal carcasses and realizing the purpose of rapid reduction treatment of the animal carcasses.
Detailed Description
The following examples are further illustrative of the present invention and are not intended to be limiting thereof.
Example 1
Fermenting and culturing bacillus stearothermophilus, bacillus amyloliquefaciens, bacillus licheniformis, bacillus calpitis, bacillus stearothermophilus, geobacillus stearothermophilus, Thermotoga maritima, Neurococcus thermophilus and archaeobacterium by using a fermentation culture medium at the culture temperature of 70-80 ℃ until a stable period;
b, mixing the bacillus stearothermophilus, the bacillus amyloliquefaciens, the bacillus licheniformis, the bacillus calpitus, the geobacillus stearothermophilus, the Thermotoga maritima, the tennis thermophilus and the archaeothermophilus according to a mass ratio of 1: 1: 1: 1: 1: 2: 2: 2 to form a composite microbial inoculum, and mixing the composite microbial inoculum with an adsorption medium according to a mass ratio of 1: 2, culturing for 24 hours under the conditions of 70-80 ℃ and 20% of relative humidity until the adsorption medium is changed from yellow to brown, namely the biological system is started, on the first day, mixing the adsorption medium containing the composite microbial inoculum after color change with the waste animal carcasses, adding and mixing the adsorption medium containing the composite microbial inoculum for three times in the morning, at the noon and at the evening according to the treatment amount of 100kg of waste animal carcasses per day of the adsorption medium containing the composite microbial inoculum, so that the composite microbial inoculum can degrade the animal carcasses in time, one-time centralized adding is avoided, the degradation effect is influenced, performing fermentation treatment at the temperature of 70-80 ℃, reducing the waste animal carcasses by about 90% after 24-hour treatment, increasing the treatment amount to 200kg on the second day, reducing the waste animal carcasses by about 90% by the same method as the first day, increasing the treatment amount to 300kg on the third day, reducing the waste animal carcasses by about 90% by the same method as the first day, after the regulation is stabilized, 300kg of animal carcasses are added into a biological system every day, the biological system is operated at the temperature of 70-80 ℃ and the humidity of 20%, a good biological film is formed in the biological system, the biological film of the composite microbial inoculum has the capability of stably degrading the animal carcasses, so that the reduction treatment of the waste animals is completed, the reduction effect reaches 90%, meanwhile, 10% of terminal products are obtained, and the terminal products are detected to have the protein content of more than or equal to 50%, the crude fat content of less than or equal to 12%, the ash content of less than or equal to 33%, the water content of less than or equal to 10%, the total amount of hydrolyzed amino acid of 40.1%, no toxic or harmful substances are found, and the animal carcasses are high-quality meat and bone meal raw materials. In order to ensure that the system can operate well for a long time, the complex microbial inoculum needs to be supplemented once a month, and 2kg of the complex microbial inoculum needs to be supplemented by taking constant-temperature heating equipment with the carcass treatment capacity of 500kg as an example.
The fermentation medium contains 5g of beef extract powder, 10g of peptone, 5g of yeast extract, 5g of sodium chloride and the balance of water per liter, and the pH value is 7.0. Mixing the above materials, sterilizing, and keeping.
The adsorption medium is prepared from porous white perlite particles and rice bran according to a mass ratio of 1: 1, mixing the mixture.
Comparative example
Culturing bacillus stearothermophilus, bacillus amyloliquefaciens, bacillus licheniformis, bacillus calpitus, geobacillus stearothermophilus, Thermotoga maritima, tennis thermophilus and archaebacteria thermophilus with a fermentation culture medium at 70-80 ℃ to a stationary phase for later use.
Weighing waste animal carcasses in advance, and mixing the following components in percentage by mass: the strain in the stationary phase is 250: the bacterial strain is added according to the proportion of 1, the bacterial strain is placed at 70-80 ℃, the weight of the bacterial strain is weighed after 24 hours of degradation, and the animal carcass reduction efficiency is obtained according to the weight reduction efficiency (weight before treatment-weight after treatment) ÷ weight before treatment multiplied by 100%. The decrement efficiencies of the bacillus stearothermophilus, the bacillus amyloliquefaciens, the bacillus licheniformis, the bacillus calpitus, the geobacillus stearothermophilus, the Thermotoga maritima, the tennis thermophilus and the archaebacteria thermophilus are 78.88%, 81.23%, 85.64%, 85.66%, 81.24%, 82.33%, 88.12% and 82.61% respectively. And the bacterial liquid of the 8 strains in the stationary phase is mixed according to the mass ratio of 1: 1: 1: 1: 1: 2: 2: 2, the treatment effect was 98.95% when the above-mentioned methods were carried out in a weight reduction test, and it was found that the combination produced a synergistic effect.
Claims (9)
1. A composite microbial inoculum for decomposing waste animal carcasses is characterized by consisting of Bacillus stearothermophilus, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus caldotena, Geobacillus stearothermophilus, Thermotoga maritima, Thermococcus thermophilus and Geobacillus thermophilus;
the mass ratio of the bacillus stearothermophilus, the bacillus amyloliquefaciens, the bacillus licheniformis, the bacillus calpitus, the geobacillus stearothermophilus, the thermotoga maritima, the tennis thermophilus and the archaebacterium thermophilus is 1: 1: 1: 1: 1: 2: 2: 2.
2. use of the composite bacterial agent of claim 1 for decomposing waste animal carcasses.
3. A method for decomposing a waste animal carcass by high-temperature aerobic fermentation, comprising the steps of:
inoculating the composite microbial inoculum of claim 1 on an adsorption medium, uniformly stirring, culturing until the adsorption medium is changed from yellow to brown, mixing the adsorption medium with waste animal carcasses, and fermenting to obtain a product, namely animal feed raw material meat and bone meal; the adsorption medium is a mixture consisting of perlite particles and rice bran.
4. The method according to claim 3, wherein the mass ratio of the composite microbial inoculum to the adsorption medium is 1: 2.
5. the method according to claim 3, wherein the temperature of the cultivation is 70 to 80 ℃ and the relative humidity is 20%.
6. The method according to claim 3, wherein the mass ratio of the waste animal carcasses to the adsorption medium containing the composite microbial inoculum after discoloration is (100-300): 1;
and adding and mixing the waste animal carcasses and the adsorption media containing the composite microbial inoculum for multiple times after the color of the waste animal carcasses is changed.
7. The method according to claim 3, wherein the temperature of the fermentation is 70 ℃ to 80 ℃.
8. The method according to claim 3, wherein the fermentation medium contains 5g of beef extract powder, 10g of peptone, 5g of yeast extract, 5g of sodium chloride and the balance water per liter, and has a pH of 7.0.
9. The method as claimed in claim 3, wherein the adsorption medium is prepared from perlite particles and rice bran according to a mass ratio of 1: 1, mixing the mixture.
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| KR20170122835A (en) * | 2015-03-16 | 2017-11-06 | 에꼴 뽈리떼끄닉 뻬데랄 드 로잔느 (으뻬에프엘) | Archaea bacteria in a feed of a bioactive animal, a method of producing the composition and a method of using the composition |
| CN104894007B (en) * | 2015-05-12 | 2019-07-05 | 安徽全民环保科技有限公司 | Compound microbial preparation, preparation method and application in treatment of animal carcasses |
| CN105218175A (en) * | 2015-08-18 | 2016-01-06 | 季维峰 | Special animalcule innocent treatment equipment and the treatment process of dying of illness of a kind of plant |
| CN105199980A (en) * | 2015-09-10 | 2015-12-30 | 山东华牧天元农牧股份有限公司 | Compounded microbial preparation and method for innocent treatment of animal carcasses |
| CN105344693B (en) * | 2015-09-24 | 2018-11-30 | 山东百德生物科技有限公司 | A kind of double acting state enzymatic hydrolysis and fermentation technique of animal derived organic matter solid phase |
| CN106365788A (en) * | 2016-08-25 | 2017-02-01 | 南京国龙生物科技有限公司 | Harmless microorganism treatment method for cadavers of livestock and poultry |
| CN107200608A (en) * | 2017-04-19 | 2017-09-26 | 南阳师范学院 | A kind of dead livestock and poultry is innoxious, recycling processing method |
| CN107912609A (en) * | 2017-11-15 | 2018-04-17 | 湖南耶啰耶兔业发展有限公司 | A kind of biology aquaculture fertilizer and preparation method thereof |
| CN111943741B (en) * | 2020-08-21 | 2022-03-11 | 四川润格生物科技有限公司 | Technical method for treating animals died of illness by enzyme method |
| CN113862245A (en) * | 2021-11-23 | 2021-12-31 | 山东百德生物科技有限公司 | Animal protein biological glycolysis agent |
| CN114478084A (en) * | 2022-03-15 | 2022-05-13 | 林其灿 | Pollution-free treatment method for biomedical wastes |
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