CN114437957A - Degradable microbial composition for environment-friendly toilet and application thereof - Google Patents
Degradable microbial composition for environment-friendly toilet and application thereof Download PDFInfo
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- CN114437957A CN114437957A CN202011223043.8A CN202011223043A CN114437957A CN 114437957 A CN114437957 A CN 114437957A CN 202011223043 A CN202011223043 A CN 202011223043A CN 114437957 A CN114437957 A CN 114437957A
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- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 37
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- 235000013957 Lactobacillus brevis Nutrition 0.000 claims abstract description 37
- 230000000593 degrading effect Effects 0.000 claims abstract description 36
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Classifications
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F11/00—Treatment of sludge; Devices therefor
- C02F11/02—Biological treatment
-
- 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
- C12N1/14—Fungi; Culture media therefor
Abstract
The invention provides a degrading microorganism composition for an environment-friendly toilet and application thereof, wherein the microorganism composition comprises the following components in parts by weight: bacillus subtilis, aspergillus niger, trichotheca rosea, lactobacillus brevis and high-temperature actinomycetes. The constructed specific degrading flora is used for treating toilet excrement and the like, can fully exert the symbiotic and synergistic effects among degrading single bacteria, realizes the rapid degradation of protein, starch, cellulose and the like in the excrement and has the characteristics of high degradation rate, short treatment period and strong adaptability. In the embodiment of the invention, 0.03-0.2% (mass-volume ratio) of the degrading bacteria is added into a feces regulating tank, the reaction time is 6-14 h, the COD reduction rate of the wastewater reaches more than 93.9%, and the deodorization effect is obvious; the method is used for treating the environmental protection toilet of the drilling crew and has good application prospect. In addition, the preparation method of the degrading bacteria is simple, low in cost and high in bacteria activity number, and is suitable for large-scale production and application.
Description
Technical Field
The invention belongs to the technical field of microbial treatment, relates to a degradable microbial composition for an environment-friendly toilet and application thereof, and particularly relates to a degradable microbial composition and a microbial preparation for the environment-friendly toilet of a drilling team as well as a preparation method and application thereof.
Background
Green construction is comprehensively developed for oil exploration, green engineering is built, green enterprises are created, and dry toilet reconstruction is promoted. The simple dry toilet is generally used by field temporary construction teams such as drilling teams, operation teams and the like under the limitation of water sources, drainage conditions and the like, excrement in a toilet excrement storage pool is easy to cause environmental pollution, the environment-friendly discharge requirement cannot be met, and disease infection can be caused.
In order to improve the damaged living environment, a drilling team is matched with a skid-mounted environment-friendly toilet, and the excrement is treated mainly by adopting the microbial technology. The microorganism treatment mode has the advantages of rapidness, high efficiency, no secondary pollution, low cost and the like, and also becomes one of the preferred technologies of the environmental protection toilet. However, most of the single strains sold in the market at present mainly comprise urban sewage treatment strains, are not suitable for special operation requirements of drilling teams, and often have the problems of large odor, long treatment time and the like in the use process.
Disclosure of Invention
In view of the above, the application provides a degradable microbial composition for an environment-friendly toilet and an application thereof, and the microbial composition provided by the invention is used for skid-mounted environment-friendly toilets and the like in drilling teams and has the remarkable advantages of high degradation efficiency, good deodorization effect, short treatment period and the like.
The invention provides a degrading microorganism composition for an environment-friendly toilet, which comprises the following components: bacillus subtilis, aspergillus niger, trichotheca rosea, lactobacillus brevis and high-temperature actinomycetes.
In the invention, the Bacillus subtilis Latin literature is named as Bacillus subtilis, the Aspergillus niger Latin literature is named as Aspergillus niger, the Trichoderma roseum Latin literature is named as Trichothecium roseum, the Lactobacillus brevis literature is named as Lactobacillus brevis, and the Thermoactinomyces daqus is named as Thermoactinomyces lactis.
In some embodiments, the bacillus subtilis collection number is CGMCC 1.15792; the preservation number of Aspergillus niger is CGMCC 3.17612; the preservation number of the trichotheca rosea is CGMCC 3.3745; lactobacillus brevis collection number CICC 6239; thermoactinomyces deposit number CICC 10681.
The analysis of the causes of the prior art problems includes: (1) the strain screening is blind, mainly takes the strain for treating the urban sewage as the main strain, and has poor pertinence; (2) the screened strains have weak environmental adaptability and short life cycle, the differences of geographic environments (inner Mongolia, Sichuan, northeast, Xinjiang and the like) and the differences of dietary structures (peppery, salt, sweet and the like) are not comprehensively considered, the performance requirements on the strains are different, and a single strain cannot adapt to the complex environment of a field well team; (3) the number of people in the well team is relatively centralized, the excrement needs to be quickly treated, the single strain treatment speed is low, and the field requirement cannot be met.
Based on above-mentioned background, to open-air well team construction operation characteristics, the cooperation is used with environmental protection lavatory to the well drilling team, realizes the quick degradation of excrement dirt, improves worker quality of life, and the microbial degradation fungus that the treatment effeciency is high, the deodorization is effectual, strong adaptability need be researched and selected out urgently.
Aiming at the characteristics of field environment at a well team, relatively centralized personnel and the like, the 5 degrading single bacteria are compounded and combined so as to rapidly and efficiently treat the excrement; the microbial degradation bacteria is a degradation microbial composition for an environment-friendly toilet and is prepared by compounding different single bacteria such as bacillus subtilis, aspergillus niger, trichotheca roseum, lactobacillus brevis and high-temperature actinomycetes. The bacillus subtilis can efficiently decompose protein, various sugars and starch in excrement, is easy to survive in severe environments such as acid and alkali, has the characteristics of strong adaptability and high degradation speed, can accelerate the proliferation of the whole compound microbial flora, can effectively inhibit the growth of pathogenic bacteria, and improves the life cycle of the flora; aspergillus niger has the characteristics of cracking macromolecular organic matters and indissolvable inorganic matters, and has good deodorization effect on feces; the trichotheca roseum can degrade the lignin, and substances generated after the decomposition and degradation of the trichotheca roseum are convenient for other microorganisms to absorb and utilize; the lactobacillus brevis mainly degrades cellulose and protein and has the characteristics of high temperature resistance and salt tolerance; the high-temperature actinomycetes mainly decompose starch and protein and generate heat, and can continuously decompose grease and cellulose substances after entering a high-temperature period. The invention utilizes the symbiotic synergistic effect of the 5 bactericides, can effectively adjust the internal flora structure to play different roles, improves the fecal sewage treatment efficiency, and further can avoid the problems of poor adaptability, short life cycle and the like of a single-component bactericide in use. In the invention, a biological degradation chain can be formed among the five degradation single bacteria, and if the chain is incomplete, the efficiency is low.
The invention provides a microbial preparation, which comprises the degrading microbial composition for the environment-friendly toilet; the microbial preparation is a solid preparation or a liquid preparation.
The embodiment of the invention provides a preparation method of a degradation microorganism solid preparation for an environment-friendly toilet, which comprises the following steps:
(1) respectively inoculating bacillus subtilis, aspergillus niger, trichotheca rosea, lactobacillus brevis and high-temperature actinomyces into respective fermentation culture media, culturing on a shaking table, and obtaining 5 kinds of microbial single-strain powder after centrifugation and freeze drying;
(2) respectively weighing 5 kinds of microbial single-bacterium powder obtained in the step (1), and mixing according to a certain mass ratio to obtain a microbial preparation of degrading bacteria for the environmental-friendly toilet in the well team.
Preferably, the microbial preparation comprises, by mass, 1 part of bacillus subtilis powder, 1-2 parts of aspergillus niger powder, 2-3 parts of trichotheca rosea powder, 1-2 parts of lactobacillus brevis powder and 2-4 parts of high-temperature actinomyces powder.
Preferably, the bacillus subtilis powder has the bacterial activity number of 108～109Per gram;
the bacterial activity of the Aspergillus niger powder reaches 109～1010Per gram;
the trichotheca rosea powder has the bacterial activity of 1011～1012Per gram;
the bacterial activity number of the lactobacillus brevis powder reaches 108～109Per gram;
the bacterial activity of the high-temperature actinomycete powder reaches 106～107Per gram.
Preferably, the trichotheca rosea powder is obtained by adopting a thiamine-containing culture medium for fermentation culture; the high-temperature actinomycete powder is obtained by fermenting and culturing a culture medium containing a malt extract.
Preferably, the fermentation culture temperature of the trichotheca pinicola and the high-temperature actinomycete is 26-38 ℃ independently; air is introduced into the culture medium in the fermentation process, and the gas-liquid volume ratio of the introduced air per minute is preferably 1-1.5.
Further, the fermentation medium of the bacillus subtilis comprises the following components: 10g of tryptone, 30g of beef extract, 4g of sodium chloride, 10g of agar and 1000mL of distilled water;
the Aspergillus niger fermentation medium comprises the following components: chopping 230g of potato, adding 1000mL of water, boiling for 30min, filtering, supplementing the filtrate to 1000mL, adding 30g of glucose and 15g of agar, dissolving and subpackaging;
the trichotheca rosea fermentation medium comprises the following components: 200g of potato extract, 20g of glucose, 3g of monopotassium phosphate, 1.5g of magnesium sulfate, 0.1g of thiamine and 15g of agar, and adjusting the pH value to 6;
the fermentation medium of the lactobacillus brevis comprises the following components: 10g of peptone, 10g of beef extract, 5g of yeast extract, 5g of glucose, 5g of sodium acetate, 2g of diamine citrate, 801 g of tween-801, 2g of dipotassium phosphate, 0.2g of magnesium sulfate heptahydrate, 0.05g of manganese sulfate heptahydrate, 20g of calcium carbonate and 1000mL of distilled water, and adjusting the pH to 6.8;
the fermentation medium of the high-temperature actinomycetes comprises the following components: glucose 4g, yeast extract 4g, malt extract 10g, CaCO32g of agar, 12g of agar and 1000mL of distilled water, and the pH is adjusted to 7.2.
Preferably, the fermentation conditions in step (1): the fermentation temperature is 26-38 ℃, the inoculation amount is 8-16% v/v, the stirring speed is 100-200 rpm, the fermentation time is 24-48 h, air is introduced into the culture medium in the fermentation process, and the gas-liquid volume ratio of the introduced air per minute is 1-1.5.
Preferably, the microbial preparation has an overall bacterial activity of 9.5 × 109More than one per gram.
The invention provides the application of the degrading microorganism composition for the environmental-friendly toilet or the microorganism preparation in fecal sewage treatment.
Preferably, the degradable microbial composition or microbial preparation for the environmental-friendly toilet is added into a regulating tank of field skid-mounted toilet feces of a drilling team according to the mass volume ratio of 0.03-0.2% for treatment; the treatment temperature is preferably 10-25 ℃, and the treatment time is 6-14 h.
Compared with the prior art, the constructed specific degradation flora is used for treating toilet excrement and the like, can fully exert the symbiotic and synergistic effects among single degradation bacteria, realizes the rapid degradation of protein, starch, cellulose and the like in the excrement, and has the characteristics of high degradation rate, short treatment period and strong adaptability. In the embodiment of the invention, 0.03-0.2% (mass-volume ratio) of the degrading bacteria is added into a feces regulating pond, the reaction time is 6-14 h, the COD reduction rate of the wastewater reaches more than 93.9%, and the deodorization effect is obvious; the method is used for treating the environmental-friendly toilet of the drilling crew and has good application prospect.
In addition, the preparation method of the degrading bacteria is simple, low in cost and high in bacteria activity number, and is suitable for large-scale production and application.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other examples, which may be modified or appreciated by those of ordinary skill in the art based on the description of the embodiments, are within the scope of the present invention.
The invention provides a degrading microorganism composition for an environment-friendly toilet, which comprises the following components: bacillus subtilis, aspergillus niger, trichotheca rosea, lactobacillus brevis and high-temperature actinomycetes.
Aiming at the technical defects of weak environmental adaptability, poor deodorization effect and short life cycle of the existing strains, the invention provides the degrading bacteria for the environmental-friendly toilet in the well team, which has strong adaptability, good deodorization effect, short treatment period and long life cycle.
In order to achieve the purpose, the degradation bacteria for the environment-friendly toilet provided by the embodiment of the invention comprise 5 different degradation single bacteria, namely bacillus subtilis, aspergillus niger, trichotheca rosea, lactobacillus brevis and high-temperature actinomyces.
The Bacillus subtilis Latin is named as Bacillus subtilis, can efficiently decompose protein, various sugars and starch in excrement, is easy to survive in severe environments such as acid and alkali, has the characteristics of strong adaptability and high degradation speed, can accelerate the proliferation of the whole compound microbial flora, can effectively inhibit the growth of pathogenic bacteria, and improves the life cycle of the flora.
The Aspergillus niger has the Latin literature name of Aspergillus niger, has the characteristic of cracking macromolecular organic matters and indissolvable inorganic matters, and has better deodorization effect on feces.
The trichotheca roseum latin literature is named as Trichotheciumureum, can degrade lignin, and substances generated after the decomposition and degradation of the lignin can be absorbed and utilized by other microorganisms conveniently.
The Lactobacillus brevis is named Lactobacillus brevis in literature, mainly degrades cellulose and protein, and has the characteristics of high temperature resistance and salt tolerance.
The high temperature actinomycete Latin literature is named as Thermoactinomyces daqus, which mainly decomposes starch and protein and generates heat, and can continuously decompose grease and cellulose substances after entering a high temperature period.
In the embodiment of the invention, the 5 degradation single bacteria are all commercially available bacteria and are preserved by China general microbiological culture preservation management center and China industrial microbiological culture preservation management center. Specifically, the preservation number of the bacillus subtilis is CGMCC 1.15792; the preservation number of Aspergillus niger is CGMCC 3.17612; the preservation number of the trichotheca rosea is CGMCC 3.3745; lactobacillus brevis deposit number CICC 6239; thermoactinomyces collection number CICC 10681.
In the embodiment of the invention, the total viable count of the bacillus subtilis, the aspergillus niger, the trichotheca roseum, the lactobacillus brevis and the high-temperature actinomyces can reach 109～1012Per gram.
The degrading bacteria provided by the invention are used for a skid-mounted environmental-friendly toilet in a well team, and have the remarkable advantages of high degrading efficiency, good deodorizing effect, short treatment period, strong adaptability and good prospect.
The embodiment of the invention provides a microbial preparation, which comprises the degradable microbial composition for the environment-friendly toilet; the microbial preparation is a solid preparation or a liquid preparation.
In the embodiment of the invention, the 5 kinds of single bacteria are respectively fermented and cultured, and are mixed according to a certain proportion to obtain the preparation of the degrading bacteria, namely the solid or liquid preparation of the degrading microorganism composition for the environment-friendly toilet.
In the specific embodiment of the present invention, bacillus subtilis, aspergillus niger, trichotheca roseum, lactobacillus brevis and thermoactinomyces are respectively inoculated in a fermentation medium, and cultured on a shaking table, preferably centrifuged and freeze-dried to obtain 5 kinds of microbial single-bacterium powders (agents) of bacillus subtilis powder, aspergillus niger powder, trichotheca rosea powder, lactobacillus brevis powder and thermoactinomyces powder.
In the method for preparing the compound microorganism bacteria disclosed by the embodiment of the invention, in order to exert the synergistic effect of mutual beneficial symbiosis among different microorganisms to the maximum extent so as to realize the rapid and efficient degradation of feces by the compound bacteria, the microorganism culture fermentation conditions and the industrial culture medium are optimized in the experimental process.
Preferably, the trichotheca pini powder is obtained by fermenting and culturing the trichotheca pini powder in a thiamine-containing culture medium. Preferably, the Thermoactinomyces powder is obtained by fermentation culture using a culture medium containing a malt extract. In a specific embodiment of the present invention, the fermentation medium of bacillus subtilis comprises: 10g of tryptone, 30g of beef extract, 4g of sodium chloride, 10g of agar and 1000mL of distilled water. The Aspergillus niger fermentation medium comprises the following components: 230g of potato is cut up, 1000mL of water is added to be boiled for 30min, the mixture is filtered, the filtrate is made up to 1000mL, 30g of glucose and 15g of agar are added, and the mixture is packaged after being dissolved.
Specifically, the trichotheca rosea fermentation medium comprises the following components: 200g of potato extract, 20g of glucose, 3g of monopotassium phosphate, 1.5g of magnesium sulfate, 0.1g of thiamine and 15g of agar, and the pH is adjusted to 6. Especially, thiamine in the culture medium has important effects on glucose metabolism, maintenance of in vivo oxidative metabolism balance and other aspects, and the activity and the degradation performance of trichotheca rosea are greatly improved.
Further, the fermentation medium of lactobacillus brevis comprises the following components: 10g of peptone, 10g of beef extract, 5g of yeast extract, 5g of glucose, 5g of sodium acetate, 2g of citric acid diamine, 801 g of tween-801, 2g of dipotassium phosphate, 0.2g of magnesium sulfate heptahydrate, 0.05g of manganese sulfate heptahydrate, 20g of calcium carbonate and 1000mL of distilled water, and adjusting the pH to 6.8.
The preferable composition of the fermentation medium of the high-temperature actinomycetes is as follows: glucose 4g, yeast extract 4g, malt extract 10g, CaCO32g of agar, 12g of agar and 1000mL of distilled water, and the pH is adjusted to 7.2. Wherein the culture medium is added with malt extract, contains abundant maltose, fructose, glucose, protein, small molecular peptide, essential amino acids, vitamins, minerals, beta-dextran and tocopherol with physiological activity function, provides comprehensive nutrition components for Thermoactinomyces, and promotes thallusThe growth is healthier, the activity of the bacteria is improved, and the comprehensive cost is reduced.
The above-mentioned conditions for culturing and fermenting microorganisms preferably include: the fermentation temperature is between 26 and 38 ℃; the inoculation amount is 8-16% v/v, the stirring speed is 100-200 rpm, and the fermentation time is 24-48 h. And in the fermentation process of each microorganism, air is introduced into the culture medium, and the gas-liquid volume ratio of the introduced air per minute is preferably 1-1.5. After fermentation culture, 5 kinds of microbial single-strain powder are obtained by conventional centrifugation and freeze drying in the embodiment of the invention.
Further, the bacillus subtilis powder has the bacterial activity number of 108～109Per gram.
Further, the bacterial activity of the Aspergillus niger powder reaches 109～1010Per gram.
Further, the trichotheca rosea powder has the bacterial activity number of 1011～1012Per gram.
Further, the bacterial activity number of the lactobacillus brevis powder reaches 108～109Per gram.
Further, the bacterial activity number of the high-temperature actinomycete powder reaches 106～107Per gram.
According to the embodiment of the invention, the 5 kinds of microbial single-bacterium powder are respectively weighed, and 1 part of bacillus subtilis powder, 1-2 parts of aspergillus niger powder, 2-3 parts of trichotheca rosea powder, 1-2 parts of lactobacillus brevis powder and 2-4 parts of high-temperature actinomyces powder are fully mixed according to the mass ratio to obtain the degrading bacteria for the environment-friendly toilet. Namely, the microbial preparation provided by the embodiment of the invention is formed by mixing the single bacterium powder; in addition, the microbial preparation can also be prepared into a liquid dosage form.
In some embodiments of the present invention, the bacillus subtilis powder, the aspergillus niger powder, the trichotheca rosea powder, the lactobacillus brevis powder and the actinomyces thermophilus powder are prepared by the following components in percentage by mass: 1: 1-2: 2-3: 1-2: 2 to 4, for example, 1:1:2:1:2, 1:2:3:2:4, 1:1.5:2.5:1.5:3, 1:1.8:2.2:1.2:3.6, etc. The total bacterial activity number of the microbial preparation is 9.5 multiplied by 109More than one per g, e.g. 9.6X 109Each/g, 4.8 is multiplied by 1010Each g, 7.9 is multiplied by 1011Each g, 6.2 x 10101.3 x 10 of seed/g12Counts/g, etc.
The embodiment of the invention provides an application of the degradable microbial composition for the environmental-friendly toilet or the microbial preparation in fecal sewage treatment, which mainly aims at a field skid-mounted toilet in a well team and specifically comprises the following steps:
adding the degradable microbial composition or microbial preparation for the environment-friendly toilet into a regulating tank for field skid-mounted toilet feces of a drilling team according to the mass volume ratio of 0.03-0.2%, fully stirring the mixed materials, and fully reacting at the temperature of 10-25 ℃ for 6-14 h.
In some embodiments of the invention, the COD value of the fecal sewage to be treated is 1768-3981 mg/L, and the total nitrogen value is 513-591 mg/L; the addition amount of the added microbial preparation is preferably 0.03-0.2% (mass/volume ratio). In some embodiments of the invention, 0.03-0.2% (mass-volume ratio) of the degrading bacteria are added into a feces regulating pond, the reaction time is 6-14 h, the COD reduction rate of the wastewater reaches more than 93.9%, and the deodorization effect is obvious; the method is used for treating the environmental protection toilet of the drilling crew and has good application prospect.
In order to further understand the present invention, the degradable microorganism composition for environmental toilet and the application thereof provided in the present application will be specifically described below with reference to the examples. It should be understood, however, that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention, which is defined by the following examples.
In the following examples, all of the 5 degradation single bacteria are commercially available bacteria, and are preserved by the China general microbiological culture Collection center and the China Industrial microbiological culture Collection center. The preservation number of the bacillus subtilis is CGMCC 1.15792, the preservation number of the aspergillus niger is CGMCC 3.17612, the preservation number of the trichotheca roseum is CGMCC3.3745, the preservation number of the lactobacillus brevis is CICC 6239 and the preservation number of the high-temperature actinomyces is CICC 10681.
The fermentation medium of the bacillus subtilis comprises the following components: 10g of tryptone, 30g of beef extract, 4g of sodium chloride, 10g of agar and 1000mL of distilled water.
The Aspergillus niger fermentation medium comprises the following components: 230g of potato is cut up, 1000mL of water is added to be boiled for 30min, the mixture is filtered, the filtrate is made up to 1000mL, 30g of glucose and 15g of agar are added, and the mixture is packaged after being dissolved.
The trichotheca rosea fermentation medium comprises the following components: 200g of potato extract, 20g of glucose, 3g of monopotassium phosphate, 1.5g of magnesium sulfate, 0.1g of thiamine and 15g of agar, and the pH is adjusted to 6.
The fermentation medium of the lactobacillus brevis comprises the following components: 10g of peptone, 10g of beef extract, 5g of yeast extract, 5g of glucose, 5g of sodium acetate, 2g of diamine citrate, 801 g of tween-801, 2g of dipotassium phosphate, 0.2g of magnesium sulfate heptahydrate, 0.05g of manganese sulfate heptahydrate, 20g of calcium carbonate and 1000mL of distilled water, and adjusting the pH to 6.8.
The fermentation medium of the high-temperature actinomycetes comprises the following components: glucose 4g, yeast extract 4g, malt extract 10g, CaCO32g of agar, 12g of agar and 1000mL of distilled water, and the pH is adjusted to 7.2.
Example 1
(1) Respectively inoculating bacillus subtilis, aspergillus niger, trichotheca rosea, lactobacillus brevis and high-temperature actinomyces into respective fermentation culture media, culturing on a shaking table, and obtaining 5 kinds of microorganism single-bacterium powder after centrifugation and conventional freeze drying;
wherein the fermentation conditions are as follows: the fermentation temperature is 26 ℃, the inoculation amount is 8% v/v, the stirring speed is 100rpm, the fermentation time is 24h, air is introduced into the culture medium in the fermentation process, and the gas-liquid volume ratio of the introduced air per minute is 1.
The bacillus subtilis powder has the bacterial activity number of 2.1 multiplied by 108Per gram;
the bacterial activity of the Aspergillus niger powder reaches 5.4 multiplied by 109Per gram;
the trichotheca rosea powder has the bacterial activity number of 7.8 multiplied by 1011Per gram;
the bacterial activity of the lactobacillus brevis powder reaches 6.6 multiplied by 108Per gram;
the number of the bacteria activity of the high temperature actinomycete powder reaches 9.0 multiplied by 106Per gram.
(2) And (2) respectively weighing 5 kinds of microbial single-bacterium powder obtained in the step (1), and fully mixing 1 part of bacillus subtilis, 1 part of aspergillus niger, 2 parts of trichotheca roseum, 1 part of lactobacillus brevis and 2 parts of high-temperature actinomycetes according to the mass ratio to obtain the degrading bacteria (namely the microbial preparation) for the environmental-friendly toilet in the well team.
Some specific preparations are listed in table 1:
TABLE 1 preparation Condition parameters of degradation bacteria for environmental protection toilets of well team
Example 2
(1) Respectively inoculating bacillus subtilis, aspergillus niger, trichotheca rosea, lactobacillus brevis and high-temperature actinomyces into a fermentation medium, culturing on a shaking table, and centrifuging, freezing and drying to obtain 5 kinds of microbial single-strain powder;
wherein the fermentation conditions are as follows: the fermentation temperature is 38 ℃, the inoculum size is 16% v/v, the stirring speed is 200rpm, the fermentation time is 48h, air is introduced into the culture medium in the fermentation process, and the gas-liquid volume ratio of the introduced air per minute is 1.5.
The bacillus subtilis powder has the bacterial activity number of 3.3 multiplied by 108Per gram;
the bacterial activity of the Aspergillus niger powder reaches 9.2 multiplied by 109Per gram;
the trichotheca rosea powder has the bacterial activity number of 7.9 multiplied by 1012Per gram;
the bacterial activity of the lactobacillus brevis powder reaches 1.8 multiplied by 108Per gram;
the bacterial activity of the high-temperature actinomycete powder reaches 6.2 multiplied by 107Per gram.
(2) And (2) respectively weighing 5 kinds of microbial single-bacterium powder obtained in the step (1), and fully mixing 1 part of bacillus subtilis, 2 parts of aspergillus niger, 3 parts of trichotheca roseum, 2 parts of lactobacillus brevis and 4 parts of high-temperature actinomycetes according to a mass ratio to obtain the degrading bacteria for the environmental-friendly toilet in the Jing team.
The embodiments 3 to 8 are similar to the methods described in the embodiments 1 to 2, and are not described herein again; some specific preparations are shown in table 1.
Comparative example 1
Similar to the method described in example 1, except that in step (2), 4 kinds of single-microorganism powder obtained in step (1) are respectively weighed and mixed, according to the mass ratio, 1 part of bacillus subtilis, 1 part of aspergillus niger, 1 part of lactobacillus brevis and 2 parts of high-temperature actinomycetes are fully mixed, and the degrading bacteria for the environmental protection toilet of the well team are obtained.
Comparative example 2
Similar to the method described in example 2, except that in step (2), 4 kinds of microbial single-bacterium powder obtained in step (1) are respectively weighed, and 1 part of bacillus subtilis, 2 parts of aspergillus niger, 3 parts of trichotheca roseum and 2 parts of lactobacillus brevis are fully mixed according to the mass ratio to obtain the degrading bacteria for the environmental protection toilet of the well team.
And (3) analyzing the application effect:
taking the waste water (the COD value of the waste water is 2897mg/L, the ammonia nitrogen content is 580mg/L) of a sedimentation tank in an environment-friendly toilet on site, adding degrading bacteria with different mass-volume ratios according to the table 2, fully stirring, reacting for different times, and measuring the COD value and the ammonia nitrogen value of the treated water.
COD was determined according to GB/T11914-.
And (4) measuring the ammonia nitrogen according to HJ535-2009 Nanshi reagent spectrophotometry for measuring the ammonia nitrogen in water. Firstly, preparing a standard storage solution, a potassium sodium tartrate solution and a Nassner reagent, then preparing an ammonium standard use solution, and drawing a standard curve of ammonia nitrogen. Centrifuging a proper amount of sample, loading the supernatant into a test tube, diluting by 10 times, loading the diluted sample solution into a colorimetric tube, calibrating to scale by using deionized water, adding a sodium potassium tartrate solution, shaking up, adding a sodium naeskowster reagent, shaking up, and standing for 10 min; and (5) measuring the absorbance of the ammonia nitrogen by using a spectrophotometer, and calculating the content of the ammonia nitrogen from the standard curve.
The results of the application analysis are as follows:
TABLE 2 evaluation of the effect of degrading bacteria
As shown in the results of Table 2, when the addition of the degrading bacteria is 0.03-0.2%, the COD degradation rate reaches 93.9-98.3% and the ammonia nitrogen degradation rate reaches 94.1-97.9% after the treatment is carried out for 6-14 h. Taking the degrading bacteria prepared in example 5 as an example, the degradation rule is shown in that as the addition of the degrading bacteria is increased, the reaction time is reduced, the reaction temperature is reduced, the COD degradation rate is basically maintained at 93.1-95.5%, and the ammonia nitrogen degradation rate is basically maintained at 94.5-97.1%. In the comparative examples 1 and 2, the addition and use of one strain are reduced, the degradation effect is obviously reduced, and the compounding synergistic effect among the strains is not fully exerted. The contrast shows that the degrading bacteria disclosed by the invention have the advantages of good COD (chemical oxygen demand) degrading effect on feces, high ammonia nitrogen degrading rate, good deodorizing effect, high degrading speed (the conventional natural degrading period is more than 3 days), no secondary pollution in the whole treating process, and environmental friendliness.
Finally, it should be noted that: although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments described in the foregoing embodiments, or equivalents may be substituted for elements thereof. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A degradable microbial composition for an environment-friendly toilet, which is characterized by comprising: bacillus subtilis, aspergillus niger, trichotheca rosea, lactobacillus brevis and high-temperature actinomycetes.
2. The environment-friendly toilet degrading microorganism composition as claimed in claim 1, wherein the Bacillus subtilis has a preservation number of CGMCC 1.15792; the preservation number of Aspergillus niger is CGMCC 3.17612; the preservation number of the trichotheca rosea is CGMCC 3.3745; lactobacillus brevis deposit number CICC 6239; thermoactinomyces deposit number CICC 10681.
3. A microbial preparation comprising the degradable microbial composition for eco-toilets according to any one of claims 1 to 2; the microbial preparation is a solid preparation or a liquid preparation.
4. The microbial preparation according to claim 3, wherein the microbial preparation comprises 1 part of bacillus subtilis powder, 1-2 parts of aspergillus niger powder, 2-3 parts of trichotheca roseum powder, 1-2 parts of lactobacillus brevis powder and 2-4 parts of Thermoactinomyces actinomyces powder in parts by mass.
5. The microbial preparation of claim 4, wherein the Bacillus subtilis powder has a viability count of up to 108～109Per gram; the bacterial activity of the Aspergillus niger powder reaches 109～1010Per gram; the trichotheca rosea powder has the bacterial activity of 1011～1012Per gram; the bacterial activity number of the lactobacillus brevis powder reaches 108～109Per gram; the bacterial activity of the high-temperature actinomycete powder reaches 106～107Per gram.
6. The microbial preparation according to claim 5, wherein the trichotheca rosea powder is obtained by fermentation culture of a thiamine-containing culture medium; the high-temperature actinomycete powder is obtained by fermenting and culturing a culture medium containing a malt extract.
7. The microbial preparation of claim 6, wherein the fermentation culture temperature of trichotheca rosea and Thermoactinomyces is 26-38 ℃ independently; air is introduced into the culture medium in the fermentation process, and the gas-liquid volume ratio of the introduced air per minute is preferably 1-1.5.
8. A microbial preparation according to any of claims 4 to 7, having an overall viability count of 9.5 x 109More than one per gram.
9. Use of the environmentally friendly lavatory degrading microbial composition of any one of claims 1 to 2 or the microbial preparation of any one of claims 3 to 8 in fecal sewage treatment.
10. The use of claim 9, wherein the degradable microbial composition or microbial preparation for the environmental-friendly toilet is added into a conditioning tank of the feces of a skid-mounted toilet in the field of a drilling crew at a mass-volume ratio of 0.03-0.2% for treatment; the treatment temperature is preferably 10-25 ℃, and the treatment time is 6-14 h.
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| CN103468614A (en) * | 2013-09-16 | 2013-12-25 | 中国人民解放军总后勤部军需装备研究所 | Kitchen waste decomposition bacterial agent and preparation method thereof |
| CN105907673A (en) * | 2016-04-29 | 2016-08-31 | 广州市佳境水处理技术工程有限公司 | Method for strengthening biochemical water treatment effects by microbial preparation |
| CN107164289A (en) * | 2017-07-18 | 2017-09-15 | 陈留成 | A kind of microorganism formulation of efficient degradation deodorization and preparation method thereof |
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| CN103468614A (en) * | 2013-09-16 | 2013-12-25 | 中国人民解放军总后勤部军需装备研究所 | Kitchen waste decomposition bacterial agent and preparation method thereof |
| CN105907673A (en) * | 2016-04-29 | 2016-08-31 | 广州市佳境水处理技术工程有限公司 | Method for strengthening biochemical water treatment effects by microbial preparation |
| CN107164289A (en) * | 2017-07-18 | 2017-09-15 | 陈留成 | A kind of microorganism formulation of efficient degradation deodorization and preparation method thereof |
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