CN112375706A - Culture method of microbial strain for degrading kitchen waste - Google Patents
Culture method of microbial strain for degrading kitchen waste Download PDFInfo
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- CN112375706A CN112375706A CN202011264099.8A CN202011264099A CN112375706A CN 112375706 A CN112375706 A CN 112375706A CN 202011264099 A CN202011264099 A CN 202011264099A CN 112375706 A CN112375706 A CN 112375706A
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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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/32—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Bacillus (G)
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Abstract
The invention is suitable for the technical field of microbial culture, and provides a culture method of microbial strains for degrading kitchen waste, which comprises the following steps: step 1, sterilizing the nutrient solution for inoculation at high temperature; step 2, cooling the inoculated nutrient solution after high-temperature sterilization; step 3, inoculating strains into the nutrient solution after cooling; select the bacterial colony that the bacterial colony becomes red more promptly and decompose the stronger bacterial colony of grease ability, continue to cultivate through the temperature interval of difference, and observe the bacterial colony at the not growth condition of temperature interval, finally select to grow vigorously, it is strong to decompose the grease ability, and at the bacterial colony of suitable temperature interval, and enlarge the culture range, finally make the fungus preparation, cultivate through the selection to the bacterial colony, the decomposition efficiency of fungus preparation to kitchen garbage has been strengthened greatly, the bacterial species of microbiological treatment technique adoption on the market at present has been avoided, efficiency is not high, and easy secondary pollution's problem.
Description
Technical Field
The invention belongs to the technical field of microbial culture, and particularly relates to a culture method of microbial strains for degrading kitchen waste.
Background
The microorganisms such as bacteria, actinomycetes and fungi widely distributed in the nature controllably promote the biochemical process of converting biodegradable organic matters into stable humus, finally form similar humus, and can be used as a fertilizer or a soil conditioner. The microbial high-temperature degradation technology is an important way for realizing the reclamation and harmlessness of the municipal waste, can kill pathogenic bacteria in the waste and effectively treat organic matters in the waste so as to finally degrade the kitchen waste into carbon dioxide, water and organic matters
The strains adopted by the microbial treatment technology in the current market have low efficiency, are treated at normal temperature, are insufficient in dehydration and exhaust, generally have low garbage reduction rate, and often cause secondary pollution of germs.
Disclosure of Invention
The invention provides a culture method of microbial strains for degrading kitchen waste, and aims to solve the problems that strains adopted by the existing microbial treatment technology in the market are low in efficiency, are treated at normal temperature, are insufficient in dehydration and exhaust, are low in waste reduction rate generally and often cause secondary pollution of germs.
The invention is realized in such a way that a culture method of microbial strains for degrading kitchen waste comprises the following steps:
step 1, sterilizing the nutrient solution for inoculation at high temperature;
step 2, cooling the inoculated nutrient solution after high-temperature sterilization;
step 3, inoculating strains into the nutrient solution after cooling;
step 4, placing the culture dish in a thermostat upside down for culture;
step 5, selecting colonies and further culturing;
step 6, preparing an oil culture medium, inoculating, and transplanting the bacterial colony into the oil culture medium for culture;
step 7, checking whether the strains can decompose the grease or not;
8, selecting bacterial colonies with strong capability of decomposing grease, and enlarging the culture range;
step 9, placing the mixture into a constant-temperature incubator for culture;
and step 10, selecting strains with strong activity to prepare a bacterium preparation.
Preferably, in the step 1, the temperature for sterilizing the inoculated nutrient solution at high temperature is 121 ℃, and the time for sterilizing at high temperature is not less than 20 minutes.
Preferably, an inner container for placing the nutrient solution is required to be arranged during cooling in the step 2, an outer container is sleeved on the inner container, a gap is reserved between the outer surface of the inner container and the inner surface of the outer container, and cooling liquid is introduced into the gap to exchange heat with the nutrient solution.
Preferably, the set temperature for cooling the nutrient solution in the step 3 is 20 to 30 degrees celsius, and preferably 25 degrees celsius.
Preferably, the strains in step 3 comprise bacillus subtilis, bacillus licheniformis and thermoactinomyces, and the content ratio of the bacillus subtilis, the bacillus licheniformis and the thermoactinomyces in the culture medium is 1:1: 1.
Preferably, in the step 3, when the strain is placed into the nutrient solution during inoculation, a circle of combustible medium is annularly arranged at an inlet where the strain is placed, and when the strain is placed, the combustible medium is ignited to form a flame sealing ring.
Preferably, in the step 7, 1.6% neutral red water solution is added to the oil culture medium, the 1.6% neutral red water solution is prepared by adding 60mL 95% ethanol and 40mL water to a neutral red reagent, and observing the culture medium after 48h to see whether the periphery of the colony is red.
Preferably, the temperature in the constant temperature incubator in the step 9 is 50 ℃ to 80 ℃, and the temperature is divided into a plurality of temperature intervals to perform control experiments respectively.
Compared with the prior art, the invention has the beneficial effects that: the invention relates to a method for culturing microbial strains for degrading kitchen waste, which comprises the steps of inoculating strains into a culture medium for culturing, selecting strains with vigorous growth, transplanting bacterial colonies into an oil culture medium for culturing for two days, adding 1.6% neutral red water solution into the oil culture medium, observing the culture medium after two days, observing whether the periphery of the bacterial colonies turns red, selecting bacterial colonies with more red bacterial colonies, namely stronger oil decomposition capacity, continuing culturing in different temperature intervals, observing the growth conditions of the bacterial colonies in different temperature intervals, finally selecting bacterial colonies with vigorous growth, strong oil decomposition capacity and proper temperature intervals, enlarging the culture range, finally preparing a bacterial preparation, greatly enhancing the decomposition efficiency of the bacterial preparation on the kitchen waste by selecting and culturing the bacterial colonies, avoiding the strains adopted by a microbial treatment technology in the current market, low efficiency and easy secondary pollution.
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FIG. 1 is a flow chart of a method of the present invention;
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Referring to fig. 1, the present invention provides a technical solution: a culture method of microbial strains for degrading kitchen waste comprises the following steps:
step 1, sterilizing the nutrient solution for inoculation at high temperature;
step 2, cooling the inoculated nutrient solution after high-temperature sterilization;
step 3, inoculating strains into the nutrient solution after cooling;
step 4, placing the culture dish in a thermostat upside down for culture;
step 5, selecting colonies and further culturing;
step 6, preparing an oil culture medium, inoculating, and transplanting the bacterial colony into the oil culture medium for culture;
step 7, checking whether the strains can decompose the grease or not;
8, selecting bacterial colonies with strong capability of decomposing grease, and enlarging the culture range;
step 9, placing the mixture into a constant-temperature incubator for culture;
and step 10, selecting strains with strong activity to prepare a bacterium preparation.
In the embodiment, firstly, the strain is inoculated into a culture medium for culture, then the strain with vigorous growth is selected, and the bacterial colony is transplanted into an oil culture medium for culture for two days, then 1.6% neutral red water solution is added into the oil culture medium, the 1.6% neutral red water solution is prepared by mixing 60mL 95% ethanol and 40mL water with a neutral red reagent, the culture medium is observed after 48 hours, whether the periphery of the bacterial colony turns red or not is observed, the bacterial colony which turns red more, namely has strong grease decomposition capability is selected, the bacterial colony is continuously transplanted into the culture medium, the growth conditions of the bacterial colony in different temperature intervals are observed, finally, the bacterial colony which grows vigorously and has strong grease decomposition capability and is in a proper temperature interval is selected, the culture range is expanded, and the bacterial preparation is finally prepared, so that the decomposition efficiency of the bacterial preparation on the kitchen waste is greatly enhanced, the problems that strains adopted by the microbial treatment technology in the current market are low in efficiency, the strains are treated at normal temperature, dehydration and exhaust are insufficient, the waste reduction rate is not high generally, and secondary pollution of germs is often caused are solved.
In the embodiment, firstly, the temperature for sterilizing the inoculated nutrient solution at high temperature is 121 ℃, the time for sterilizing at high temperature is not less than 20 minutes, other bacteria in the nutrient solution can be effectively killed, the pollution of the other bacteria and fungi to strains is avoided, an inner container for placing the nutrient solution is arranged after sterilizing at high temperature, an outer container is sleeved on the inner container, a gap is reserved between the outer surface of the inner container and the inner surface of the outer container, then cooling liquid is introduced into the gap to exchange heat with the nutrient solution, the cooling of the nutrient solution is accelerated, after the nutrient solution is cooled to 20 ℃ to 30 ℃, bacillus subtilis, bacillus licheniformis and high-temperature actinomycetes with the content ratio of 1:1:1 are inoculated into a culture medium, when the strains are placed into the nutrient solution, a circle of combustible medium is arranged at an inlet for placing the strains, when the strains are placed, igniting a combustible medium to form a flame seal ring, preventing external bacteria from entering a culture medium during inoculation and polluting strains inside the culture medium, after the inoculation is finished, placing a culture dish inside a 30-DEG C constant temperature box for culturing for two days, selecting colonies with vigorous growth, further culturing, manufacturing an oil culture medium after the culture is finished, transplanting the colonies after the further culture into the oil culture medium, placing the oil culture medium inside the 30-DEG C constant temperature box for culturing for two days, adding 1.6% of neutral red water solution into the oil culture medium, adding 60mL 95% of ethanol and 40mL of water into the 1.6% of neutral red water solution, observing the culture medium after two days, observing whether the culture medium turns red around the colonies, selecting the colonies with more red color, namely further culturing the colonies with stronger grease decomposition capacity, and culturing the colonies at a temperature of 50-80 ℃ according to an interval of every five DEG C, and selecting the temperature interval which is most vigorous in growth and most sufficient in grease decomposition, expanding the culture range, finally preparing a bacterial preparation, and through selective cultivation of bacterial colonies, greatly enhancing the decomposition efficiency of the bacterial preparation on the kitchen waste, avoiding the problems that the bacterial strain adopted by the microbial treatment technology in the current market is low in efficiency, is treated at normal temperature, is insufficient in dehydration and exhaust, is not high in waste reduction rate generally and often causes secondary pollution of pathogenic bacteria.
Further, in the step 1, the temperature for sterilizing the inoculated nutrient solution at high temperature is 121 ℃, and the time for sterilizing at high temperature is not less than 20 minutes.
In the embodiment, the temperature for sterilizing the inoculated nutrient solution at high temperature is 121 ℃, and the time for sterilizing at high temperature is not less than 20 minutes, so that other bacteria in the nutrient solution can be effectively killed, and the pollution of other bacteria and fungi to strains is avoided.
Furthermore, an inner container for placing nutrient solution is required to be arranged during cooling in the step 2, an outer container is sleeved on the inner container, a gap is reserved between the outer surface of the inner container and the inner surface of the outer container, and cooling liquid is introduced into the gap to exchange heat with the nutrient solution.
In the present embodiment, the nutrient solution is placed inside the inner container, the outer container is fitted around the inner container, and a gap is provided between the inner container and the outer container to fill a proper amount of the coolant into the gap, thereby accelerating the cooling process of the nutrient solution inside the inner container.
Further, the set temperature for cooling the nutrient solution in step 3 is 20 to 30 degrees celsius, preferably 25 degrees celsius.
In the embodiment, the nutrient solution is cooled to 20-30 ℃ and attached to the natural temperature, so that the strain can be conveniently inoculated, and the selection of 25 ℃ is most beneficial to the growth and reproduction of the strain.
Further, the strains in the step 3 comprise bacillus subtilis, bacillus licheniformis and high-temperature actinomyces, and the content ratio of the bacillus subtilis, the bacillus licheniformis and the high-temperature actinomyces in the culture medium is 1:1: 1.
In the embodiment, the bacillus subtilis, the bacillus licheniformis and the high-temperature actinomyces can synergistically promote the effect, and can effectively degrade organic substances in household garbage, particularly kitchen garbage, so that the odor of the garbage can be obviously reduced, and the living environment can be improved.
Further, in step 3, when the strain is placed into the nutrient solution during inoculation, a circle of combustible medium is annularly arranged at the position of the strain placing inlet, and when the strain is placed, the combustible medium is ignited to form a flame sealing ring.
In this embodiment, when inoculating, when putting into the bacterial strain in the nutrient solution, set up the round combustible medium at the entrance ring of putting into the bacterial strain, when putting into the bacterial strain, light combustible medium, form the flame seal circle for outside bacterium or fungi get into the nutrient solution inside when preventing to inoculate, cause the bacterial colony by the problem of other bacterial contamination.
Further, in the test in step 7, 1.6% neutral red water solution is added to the oil culture medium, the 1.6% neutral red water solution is prepared by adding 60mL 95% ethanol and 40mL water to neutral red reagent, and the culture medium is observed after 48 hours to see whether the periphery of the colony is red.
In this embodiment, a 1.6% neutral red aqueous solution is added to the oil or fat medium. The 1.6% neutral red water solution is prepared by mixing 60mL 95% ethanol and 40mL water with neutral red reagent, wherein the 1.6% neutral red water solution can react with fatty acid after oil and fat decomposition to generate red color, if red colony is generated, the colony can decompose oil and fat, and if no red colony is generated, the colony can not decompose oil and fat.
Further, the temperature in the constant temperature incubator in the step 9 is 50 ℃ to 80 ℃, and the temperature is divided into a plurality of temperature ranges to perform control experiments respectively.
In this embodiment, set up a plurality of control temperature intervals of 50 degrees centigrade to 80 degrees centigrade for observe the active degree of bacterial colony in every temperature interval and to the speed of grease decomposition, thereby find out suitable temperature interval and carry out effective quick degradation to kitchen garbage.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (8)
1. A culture method of microbial strains for degrading kitchen waste is characterized by comprising the following steps: the method comprises the following steps:
step 1, sterilizing the nutrient solution for inoculation at high temperature;
step 2, cooling the inoculated nutrient solution after high-temperature sterilization;
step 3, inoculating strains into the nutrient solution after cooling;
step 4, placing the culture dish in a thermostat upside down for culture;
step 5, selecting colonies and further culturing;
step 6, preparing an oil culture medium, inoculating, and transplanting the bacterial colony into the oil culture medium for culture;
step 7, checking whether the strains can decompose the grease or not;
8, selecting bacterial colonies with strong capability of decomposing grease, and enlarging the culture range;
step 9, placing the mixture into a constant-temperature incubator for culture;
and step 10, selecting strains with strong activity to prepare a bacterium preparation.
2. The method for culturing the microbial strains for degrading the kitchen waste, according to claim 1, is characterized in that: in the step 1, the temperature for sterilizing the inoculated nutrient solution at high temperature is 121 ℃, and the time for sterilizing at high temperature is not less than 20 minutes.
3. The method for culturing the microbial strains for degrading the kitchen waste, according to claim 1, is characterized in that: and 2, an inner container for placing nutrient solution is required to be arranged during cooling in the step 2, an outer container is sleeved on the inner container, a gap is reserved between the outer surface of the inner container and the inner surface of the outer container, and cooling liquid is introduced into the gap to exchange heat with the nutrient solution.
4. The method for culturing the microbial strains for degrading the kitchen waste, according to claim 1, is characterized in that: and 3, cooling the nutrient solution in the step 3 at a set temperature of 20-30 ℃, preferably 25 ℃.
5. The method for culturing the microbial strains for degrading the kitchen waste, according to claim 1, is characterized in that: the strains in the step 3 comprise bacillus subtilis, bacillus licheniformis and high-temperature actinomycetes, and the content ratio of the bacillus subtilis, the bacillus licheniformis and the high-temperature actinomycetes in the culture medium is 1:1: 1.
6. The method for culturing the microbial strains for degrading the kitchen waste, according to claim 1, is characterized in that: in the step 3, when the strain is put into the nutrient solution during inoculation, a circle of combustible medium is annularly arranged at an inlet where the strain is put, and when the strain is put, the combustible medium is ignited to form a flame sealing ring.
7. The method for culturing the microbial strains for degrading the kitchen waste, according to claim 1, is characterized in that: when the test in the step 7 is carried out, 1.6% neutral red water solution is added into the oil culture medium, the 1.6% neutral red water solution is prepared by adding 60mL 95% ethanol and 40mL water into a neutral red reagent, and the culture medium is observed after 48 hours to see whether the periphery of the bacterial colony turns red.
8. The method for culturing the microbial strains for degrading the kitchen waste, according to claim 1, is characterized in that: and 9, controlling the temperature in the constant-temperature incubator in the step 50-80 ℃ in a plurality of temperature intervals to perform control experiments respectively.
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| CN202011264099.8A CN112375706A (en) | 2020-11-12 | 2020-11-12 | Culture method of microbial strain for degrading kitchen waste |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107164231A (en) * | 2017-07-12 | 2017-09-15 | 浙江桃花源环保科技有限公司 | The screening technique of high-efficiency grease degradation bacteria |
| WO2018133411A1 (en) * | 2017-01-17 | 2018-07-26 | 广州市广深环保科技有限公司 | Multifunctional and efficient sewage microorganism activated bacterial agent and use thereof |
-
2020
- 2020-11-12 CN CN202011264099.8A patent/CN112375706A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018133411A1 (en) * | 2017-01-17 | 2018-07-26 | 广州市广深环保科技有限公司 | Multifunctional and efficient sewage microorganism activated bacterial agent and use thereof |
| CN107164231A (en) * | 2017-07-12 | 2017-09-15 | 浙江桃花源环保科技有限公司 | The screening technique of high-efficiency grease degradation bacteria |
Non-Patent Citations (2)
| Title |
|---|
| 戚薇等: "高效降解泔脚垃圾微生物制剂的研制", 《天津科技大学学报》 * |
| 王继华等: "低温高效油脂降解菌的分离筛选驯化", 《微生物学通报》 * |
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Effective date of registration: 20211122 Address after: No.50, xinganquan East Road, Yangzhou City, Jiangsu Province, 225000 Applicant after: YANGZHOU JINWEI ENVIRONMENTAL PROTECTION TECHNOLOGY Co.,Ltd. Address before: 225000 Building 1, No.50, East xinganquan Road, Hanjiang District, Suzhou City, Jiangsu Province Applicant before: Yangzhou Xinwei environmental protection Biotechnology Co.,Ltd. |
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Application publication date: 20210219 |