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

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

Compound bacterial agent for decomposing waste animal carcasses and its application, high temperature aerobic fermentation Method for decomposing waste animal carcasses

This application is a divisional application with an application date of August 6, 2014, an application number of 201410384531.5, and the title of the invention "A Method for Decomposing Waste Animal Carcasses by High-temperature Aerobic Fermentation".

technical field

The invention belongs to the field of disposal of waste animal carcasses, in particular to a compound bacterial agent for decomposing waste animal carcasses and applications thereof, and a method for decomposing waste animal carcasses by high-temperature aerobic fermentation.

Background technique

With the rapid development of livestock and poultry breeding industry, slaughtering industry and meat processing enterprises, the waste animal carcasses produced have caused huge pressure on environmental protection. According to the "Animal Epidemic Prevention Law of the People's Republic of China": At present, the methods that can harmless, reduce, recycle, and treat waste animal carcasses include sanitary landfill, composting and incineration. From the perspective of technical process, the goals pursued by each method are the same, but each has its own advantages and disadvantages, and the degree of technical difficulty varies greatly:

(1) Incineration method; this method will produce secondary pollution, and the highly toxic three-induced (carcinogenic, teratogenic, mutagenic) gases are harmful to human health.

(2) High temperature and high pressure method; this method uses high temperature and high pressure equipment to process animal carcasses. This equipment belongs to special equipment and needs to be specially equipped with boiler workers. The waste water and waste gas generated by it need to be reprocessed with professional equipment to meet environmental protection standards, and The product of this method needs solid-liquid separation, and a separation device needs to be added.

(3) Sanitary burial; this method occupies a large area and occupies land permanently. The decay of the body during landfill will cause bubbles to emerge and liquid to leak, and the chemicals used in landfill (quick lime and bleaching powder) are harmful to groundwater. pose a greater security risk.

(4) The traditional fermentation method, which uses a mixture of rice bran, sawdust, straw, corncob and other mixtures to make compost fermentation, usually takes 1 to 3 months to complete, the time period is long, land resources are occupied for a long time, the fermentation process produces odor pollution, fermentation The product is organic fertilizer, and the economic benefit of organic fertilizer is poor.

High temperature bacteria waste animal carcass treatment technology is currently a promising organic waste treatment technology. High temperature bacteria treatment technology of waste animal carcasses refers to the use of bacteria widely distributed in nature, Bacillus, Thermotoga marina, Archaea and other microorganisms or artificial inoculants, under artificial control conditions, to ensure a certain amount of moisture and oxygen supply , temperature, etc., a method for biodegrading waste animal carcasses into meat and bone meal as animal feed raw materials. The main control measures and approaches: (1) Due to the implementation of large-scale factory farming, slaughtering and processing, the consumption of animal carcasses such as livestock is large. This method has fast response speed, high processing efficiency, and less land resources; (2) ) The waste animal carcasses are made into high-quality animal feed raw material meat and bone meal, so that the energy and nutrients in it can be recycled, and the degree of resource utilization is high; (3) The waste animal carcasses are fermented into animal feed raw material meat and bone meal, which can prevent waste animals from impersonating at the source. Fresh animal meat is sold in the market, which ensures our food safety and the resources can be recycled. (4) The high-temperature bacteria waste animal treatment technology not only solves the treatment of waste animals, but also causes pathogenic microorganisms and viruses to be killed after the resulting animal feed raw materials are fermented at high temperature for a long time, reducing the pollution of bacterial metabolites to products and ensuring resources. Use safety of chemical products (see Appendix 2, Salmonella Analysis Report of the Feed Quality Supervision, Inspection and Testing Center (Guangzhou), Ministry of Agriculture) (see Appendix 3, PRRS Analysis and Test Report of the Diagnosis Center of the Institute of Animal Health, Guangdong Academy of Agricultural Sciences) . (5) After high temperature fermentation, waste animals are completely decomposed into amino acids that are easy for animals to digest and absorb, and the total amount of hydrolyzed amino acids exceeds 40% (see Appendix 1, Analysis Report of Hydrolyzed Amino Acids by Guangzhou Analysis and Testing Center, China), and feeds made of this fermentation product It has the characteristics of strong palatability and good taste. The ability of microorganisms to decompose discarded animal carcasses is amazing. It can be said that almost all waste animals that exist in nature can be decomposed by microorganisms. Some species, such as Thermotoga spp., can degrade fat, protein, and cellulose, and can use any of these organisms as the sole carbon source for metabolism. The key to the application of high-temperature bacteria in the reduction, recycling, and harmless treatment of waste animal carcasses lies in several major inventions such as screening, separation, selection, combined culture and biofilm immobilization.

SUMMARY OF THE INVENTION

The object of the present invention is to provide a compound bacterial agent for decomposing waste animal carcasses and its application, and a method for decomposing waste animal carcasses by high-temperature aerobic fermentation, so as to achieve effective reduction, resource utilization, and harmlessness, while also decomposing waste products. Animal carcasses to produce meat and bone meal, an excellent raw material for animal feed.

The method for decomposing waste animal carcasses by high-temperature aerobic fermentation of the present invention is characterized in that, comprising the following steps:

Inoculate Bacillus stearothermophilus, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus thermophilus, Geobacillus stearothermophilus, Thermotoga marina, Tennis thermophilus, and thermophilic archaea on the adsorption medium, and stir After homogenization, cultivate until the adsorption medium changes from yellow to brown, and then mix it with discarded animal carcasses for fermentation. The fermented product is meat and bone meal, which is a raw material for animal feed. The adsorption medium is composed of perlite particles and rice bran. mixture.

The method for decomposing waste animal carcasses by high-temperature aerobic fermentation of the present invention is preferably as follows: Bacillus stearothermophilus, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus thermophilus, Bacillus thermophilus, Bacillus stearothermophilus, Bacillus stearothermophilus, Bacillus licheniformis, Bacillus thermophilus, Bacillus thermophilus, and Bacillus stearothermophilus, which are fermented and cultured in a fermentation medium to a stable stage, are preferably used in the present invention. Geobacillus, Thermotoga marina, Tennis thermophilus, and thermophilic archaea are mixed uniformly in a mass ratio of 1:1:1:1:1:2:2:2 to form a compound bacterial agent, and then compound the After the inoculum and the adsorption medium are mixed evenly according to the mass ratio of 1:2, they are incubated at 70°C to 80°C and the relative humidity is 20% until the adsorption medium changes from yellow to brown. The adsorption medium is mixed with discarded animal carcasses, and the amount of discarded animal carcasses is 100-300 kg per kilogram of the adsorption medium containing the compound bacterial agent per day. Add and mix in several times, and ferment at a temperature of 70°C to 80°C until the fermentation is completed. , the fermented product is the meat and bone meal of animal feed raw materials;

The fermentation medium contains 5 g of beef extract powder, 10 g of peptone, 5 g of yeast extract, 5 g of sodium chloride, and the balance of water, and the pH is 7.0 per liter. Sterilize and sterilize.

The adsorption medium is a product obtained by mixing perlite particles and rice bran in a mass ratio of 1:1.

Bacillus stearothermophilus (Bacillus stearothermophilus), milky white, rod-shaped, Gram-positive, with spores, the optimum growth temperature is 70 ℃ ~ 80 ℃. The strain belongs to the strain in the prior art and can be purchased from biological companies or preservation institutions, such as Bacillus stearothermophilus CICIM B0553.

Bacillus amyloliquefaciens (Bacillus amyloliquefaciens), milky white, rod-shaped, Gram-positive, with spores, the optimum growth temperature is 70 ℃. The strain belongs to the strain in the prior art and can be purchased from biological companies or preservation institutions, such as Bacillus amyloliquefaciens CICIMB4311 of the Industrial Microbial Data Platform of Chinese Universities.

Bacillus licheniformis, milky white, rod-shaped, Gram-positive, with spores, the optimum growth temperature is 70 °C. The strain belongs to the strain in the prior art and can be purchased from a biological company or a preservation institution, such as Bacillus licheniformis CICIM B1455 of the Industrial Microbial Data Platform of Chinese Universities.

Bacillus thermoleovorans is milky white, rod-shaped, Gram-positive, with spores, and the optimum growth temperature is 70℃～75℃. The strain belongs to the strain in the prior art and can be purchased from biological companies or preservation institutions, such as Bacillus thermoleovorans CICIM B0614 of the Industrial Microorganism Data Platform of Chinese Universities.

Geobacillus stearothermophilus (Geobacillus spp.) is milky white, rod-shaped, Gram-positive, with spores, and the optimum growth temperature is 70°C. The strain belongs to the strain in the prior art and can be purchased from a biological company or a preservation institution, such as Geobacillus stearothermophilus with the preservation number of GIM1.543 of the Guangdong Provincial Microbial Culture Collection Center.

Thermotoga (Thermotogaspp.), yellow-white, round, with a special sheath-like structure, the optimum growth temperature is 80 ℃. The strain belongs to the strains in the prior art (such as the paper: Cloning and expression of the xynB gene of Thermotoga mari, purification of recombinant xylanase and its enzymatic properties, Jiang Zhengqiang, China Agricultural University, published in 2001), It can be purchased from a biological company or a depository institution. The applicant also holds the strain, which is guaranteed to be available to the public within 20 years from the date of application.

Dictyoglomus thermophilum is milky white and filamentous, and its optimum growth temperature is 90°C. The strain belongs to the strain in the prior art, which is disclosed in various documents and can be purchased from biological companies or depository institutions.

Thermophilic archaea (Pyrococcus furiosus) milky white, rod-shaped, the optimum growth temperature is 80 ℃. This strain belongs to the strains in the prior art (such as the paper: Prokaryotic expression and enzymatic property analysis of lactase gene derived from thermophilic archaea Pyrococcus furiosus, Duan Wenjuan, Yang Jiaoyan, Li Zhuofu, Zhang Wei, Zhang Zhifang, China Agricultural Science and Technology Herald, 2008 (2 ): 76-81 have been published), which can be purchased from biological companies or preservation institutions. The applicant also holds this strain, and it is guaranteed that it will be available to the public within 20 years from the date of application.

(1) Determination of genetic stability, the above-mentioned 8 high-temperature aerobic fermentation bacteria can be genetically stable within 15 generations.

(2) Antibiotic Sensitivity Determination 8 kinds of antibiotics sensitive to high temperature aerobic fermentation bacteria include chloramphenicol, tetracycline, penicillin G, cefotaxime (Cef I), gentamicin, ofloxacin (ofloxacin), Polymyxin B, Clindamycin (Clindamycin), Ketoconazole, Miconazole/Dacronine and Clotrimazole.

(3) The acute skin irritation of rabbits was measured, and the acute skin irritation of 8 kinds of high-temperature aerobic fermentation bacteria to rabbits was mild.

(4) Determination of acute oral toxicity in mice, the measurement results of acute oral toxicity in mice of 8 kinds of high-temperature aerobic fermentation bacteria were >2000 mg/kg. The above results all show that the 8 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 will not be harmful to humans and animals. adverse effects on the living environment.

The present invention selects 8 kinds of bacteria, which are mixed according to a certain proportion to form a composite bacterial agent, and the composite bacterial agent forms a bacterial biofilm on the adsorption medium. The rapid degradation of waste animal carcasses under high temperature solves the problem of reducing and recycling waste animal carcasses, and realizes the purpose of rapidly reducing and disposing of animal carcasses.

Detailed ways

The following examples are further illustrations of the present invention, rather than limitations of the present invention.

Example 1

Use fermentation medium to ferment and culture Bacillus stearothermophilus, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus thermophilus, Geobacillus stearothermophilus, Thermotoga marina, Tennis thermophilus, and thermophilic archaea. The temperature is 70 ~ 80 ℃, to the stable period;

The stable phase of Bacillus stearothermophilus, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus thermophilus, Geobacillus stearothermophilus, Thermotoga marina, Tennis thermophilus, and thermophilic archaea were in a mass ratio of 1 : 1: 1: 1: 1: 2: 2: 2 is mixed to form a composite bacterial agent, and then the composite bacterial agent and the adsorption medium are mixed evenly according to the mass ratio of 1: 2, at 70 ℃ ~ 80 ℃, relatively Incubate for 24 hours under the condition of humidity of 20%, until the adsorption medium changes from yellow to brown, that is, the biological system is started. The adsorption medium of the agent is 100kg of waste animal carcasses per day, and it is added and mixed three times in the morning, middle and evening, so that the compound bacteria agent can degrade the animal carcasses in time, avoiding a centralized dose and affecting the degradation effect. The temperature is 70 ℃ ~ 80 ℃. Fermentation treatment, after 24 hours of treatment, the amount of discarded animal carcasses was reduced by about 90%, and the treatment amount was increased to 200kg on the second day. The treatment capacity was increased to 300kg. In the same way as the first day, the amount of discarded animal carcasses was reduced by about 90%. After the debugging reached stability, 300kg of animal carcasses were added to the biological system every day. % conditions, due to the formation of a good biofilm in the biological system, the compound biofilm has the ability to stably degrade animal carcasses, so as to complete the reduction of waste animals, the reduction effect reaches 90%, and at the same time, 10% of the biofilm is obtained. The end product, the end product has been tested, and the protein content is ≥ 50%, the crude fat content is ≤ 12%, the ash content is ≤ 33%, the moisture content is ≤ 10%, the total amount of hydrolyzed amino acids is 40.1%, no toxic and harmful substances are found, and it is a high-quality animal feed Raw meat and bone meal. In order to make the system run well for a long time, the compound bacterial agent needs to be supplemented once a month. Taking the constant temperature heating equipment with an animal carcass processing capacity of 500kg as an example, the compound bacterial agent needs to be supplemented with 2kg.

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, and the pH is 7.0 per liter. Mix the above substances evenly and sterilize them for later use.

The adsorption medium is a product obtained by mixing porous white perlite particles and rice bran in a mass ratio of 1:1.

Comparative ratio

Fermentation medium of Bacillus stearothermophilus, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus thermophilus, Geobacillus stearothermophilus, Thermotoga marina, Tennis thermophilus, and thermophilic archaea at 70～ Cultivated at 80 degrees Celsius to the stable phase and set aside.

The discarded animal carcasses were weighed in advance and added to the strains according to the mass ratio of animal carcasses: bacterial species in the above stable period = 250:1, placed at 70 to 80 ° C, and weighed after degrading for 24 hours. Dose efficiency=(weight before treatment-weight after treatment)÷weight before treatment×100%, that is, the reduction efficiency of animal carcasses is obtained. The reduction efficiency of Bacillus stearothermophilus, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus thermophilus, Geobacillus stearothermophilus, Thermotoga marina, Tennis thermophilus, and thermophilic archaea reached 78.88%, respectively , 81.23%, 85.64%, 85.66%, 81.24%, 82.33%, 88.12%, 82.61%. The bacterial liquids of these 8 strains in the stable stage were combined according to the mass ratio of 1:1:1:1:1:2:2:2, and the weight reduction test was carried out according to the above method, and the treatment effect was 98.95%. It can be seen that there is a synergistic effect after the combination.

Claims (9)

1. a compound microbial inoculant for decomposing abandoned animal carcasses, is characterized in that, by Bacillus stearothermophilus, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus thermophilus, Geobacillus stearothermophilus, Thermus mari Composed of Toga, Tennis Thermophilus and Thermophilic Archaea; The mass ratio of the Bacillus stearothermophilus, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus thermophilus, Geobacillus stearothermophilus, Thermotoga marina, Tennis thermophilus and thermophilic archaea is 1: 1:1:1:1:2:2:2. 2. the application of the compound bacterial agent of claim 1 in decomposing waste animal carcasses. 3. a method for decomposing waste animal carcasses by high temperature aerobic fermentation, is characterized in that, comprises the following steps: The compound bacterial agent of claim 1 is inoculated on the adsorption medium, and after stirring, culture is carried out until the adsorption medium is changed from yellow to brown, then it is mixed with discarded animal carcasses and fermented, and the fermented product is the animal feed raw material. Meat and bone meal; the adsorption medium is a mixture of perlite particles and rice bran. 4. The method according to claim 3, wherein the mass ratio of the composite bacterial agent to the adsorption medium is 1:2. 5 . The method according to claim 3 , wherein the culture temperature is 70° C.˜80° C. and the relative humidity is 20%. 6 . 6 . The method according to claim 3 , wherein the mass ratio of the discarded animal carcass to the adsorption medium containing the composite bacterial agent after discoloration is (100-300): 1; 6 . The discarded animal carcasses and the discolored adsorption medium containing the compound bacterial agent are added and mixed in multiple times. 7 . The method according to claim 3 , wherein the fermentation temperature is 70°C to 80°C. 8 . 8. method according to claim 3, is characterized in that, the substratum that every liter of described fermentation contains beef extract powder 5g, peptone 10g, yeast extract 5g, sodium chloride 5g and surplus is water, and pH is 7.0 . 9 . The method according to claim 3 , wherein the adsorption medium is a product obtained by mixing perlite particles and rice bran in a mass ratio of 1:1. 10 .