CN112479391A - Preparation method of degradation product of well site environment-friendly toilet and method for treating water-based solid waste - Google Patents
Preparation method of degradation product of well site environment-friendly toilet and method for treating water-based solid waste Download PDFInfo
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Classifications
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09B—DISPOSAL OF SOLID WASTE NOT OTHERWISE PROVIDED FOR
- B09B3/00—Destroying solid waste or transforming solid waste into something useful or harmless
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
- B09B—DISPOSAL OF SOLID WASTE NOT OTHERWISE PROVIDED FOR
- B09B5/00—Operations not covered by a single other subclass or by a single other group in this subclass
-
- 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
- C02F11/12—Treatment of sludge; Devices therefor by de-watering, drying or thickening
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/005—Black water originating from toilets
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- Engineering & Computer Science (AREA)
- Environmental & Geological Engineering (AREA)
- Life Sciences & Earth Sciences (AREA)
- Hydrology & Water Resources (AREA)
- Water Supply & Treatment (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Microbiology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Purification Treatments By Anaerobic Or Anaerobic And Aerobic Bacteria Or Animals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Treatment Of Sludge (AREA)
Abstract
The invention provides a preparation method of well site environment-friendly toilet degradation products and a method for treating water-based solid waste, wherein the method for biologically treating the water-based drilling solid waste by the cooperation of the well site environment-friendly toilet degradation products comprises the following steps: uniformly mixing water-based drilling solid waste to be treated, inoculating 0.1-0.3% of microbial degradation strain with degradation effect according to the weight ratio, and fully and uniformly mixing; adding environment-friendly toilet degradation products with the solid waste amount of 0.5-1 wt% of the water-based drilling well to be treated, and fully and uniformly mixing; adding relatively fine fresh soil with the solid waste amount of 0.1-1 time of that of the to-be-treated soil, uniformly mixing and stacking; fresh soil with a certain thickness is covered on the surface of the treated object, ornamental or firewood plants are sown and planted on the fresh soil to form a well site solid waste-microorganism-plant combined degradation system. The invention has the advantages of realizing the cooperative disposal and resource soil utilization of the drilling life solid waste and the drilling water-based solid waste, achieving the purposes of double emission reduction and utilization, realizing the principle of reduction, harmlessness and resource of the solid waste disposal, reducing the external transportation and disposal amount and cost, and the like.
Description
Technical Field
The invention belongs to the technical field of pollution treatment of petroleum and natural gas exploration drilling, and particularly relates to a preparation method of a well site environment-friendly toilet degradation product and a method for treating water-based solid waste.
Background
The water-based drilling solid waste is a necessary product of conventional drilling operation of oil and gas exploration, and the production amount in Chongqing areas is generally 0.35-0.4 m3Advancing to the length per meter. At present, in the Yu district, a small part of water-based drilling solid waste is recycled by adopting biological treatment, and most of the water-based drilling solid waste is mainly sent to a local brick factory for brick making utilization, so that the transportation treatment amount is large, the treatment cost is high, and the safety and environmental protection risks in transportation are increased. The existing circulating water flushing type microbial degradation environment-friendly toilet and the flushing-free biodegradable environment-friendly toilet are popularized and applied in drilling well sites of Chuan Yu and Changqing areas of Chuan Qing company, a certain amount of domestic solid waste is generated after the two environment-friendly toilets are used, at present, the domestic solid waste is treated in a mode of transporting organic fertilizer plants outside and delivering the organic fertilizer plants to urban sanitation for treatment or burying on the spot, and the treatment technologies have the defects of high transportation cost, post-treatment cost, high treatment cost, serious region limitation, high environmental safety risk and the like. Relevant researches show that the solid waste of the environment-friendly toilet has high contents of organic matters, nitrogen, phosphorus and potassium, and is a high-quality raw material for organic fertilizer and soil fertilization. So that the domestic solid waste generated by the environment-friendly toilet can be degraded and harmlessly treated, and the obtained degradation product is used as an organic nutrient to be synergisticThe method is used for biologically treating water-based drilling solid waste and recycling soil, so that the cooperative treatment of the environment-friendly toilet domestic solid waste and the drilling water-based solid waste and the recycling soil utilization can be realized, the purposes of dual emission reduction and utilization are achieved, the reduction, harmlessness and recycling principles of solid waste treatment are met, and the national energy-saving and emission reduction policies, ecological management requirements and the requirements of group companies on enhancing the ecological environment protection are met.
Disclosure of Invention
The present invention aims to address at least one of the above-mentioned deficiencies of the prior art. For example, the invention aims to provide a preparation method of a wellsite environment-friendly toilet degradation product and a method for cooperatively biologically treating water-based drilling solid waste by using the environment-friendly toilet degradation product.
In order to achieve the above objects, one of the objects of the present invention is to provide a method for degrading domestic wastewater and human excrement generated in an environment-friendly toilet for a wellsite.
The invention provides a preparation method of a degradation product of a wellsite environment-friendly toilet, which comprises the following steps: treating domestic wastewater and human excrement generated by the well site environment-friendly toilet by using a biological agent, and filtering and drying the treated domestic wastewater and human excrement until the water content is 30-55% to obtain a well site environment-friendly toilet degradation product; wherein the biological agent is Bacillus belgii B3-4 or its suspension or its culture solution or its fermentation product.
In one exemplary embodiment of an aspect of the present invention, the eco-friendly toilet may be a flush-free eco-friendly toilet.
In an exemplary embodiment of one aspect of the invention, the Bacillus belgii is a gram-positive bacterium, has spores, has a rod-shaped, motile, obligately aerobic body.
In an exemplary embodiment of one aspect of the invention, the 16S rDNA sequence of bacillus belgii is as shown in SEQ ID No. 1.
In an exemplary embodiment of one aspect of the invention, the colony formed by culturing the bacillus belgii on a beef extract peptone medium for 24 hours is round or irregular, and the colony after 48 hours is round and white, has the diameter of 0.5-1 mm, irregular edges and is flat and wet.
In an exemplary embodiment of one aspect of the present invention, the method for screening and culturing Bacillus belgii comprises the steps of: collecting and storing samples at low temperature, wherein the samples comprise human excrement, livestock and poultry excrement, soil, water, sludge and organic fertilizer; coating the diluent of the sample on a beef extract peptone culture medium by adopting a gradient dilution method, and culturing at a constant temperature of 15-50 ℃; after the culture medium grows out of the bacteria, selecting bacterial colonies with different forms from a beef extract peptone flat plate for streak culture; subsequently, a loop of colonies was picked for repeated streaking with microscopic observation until a purified strain was obtained.
The invention also aims to provide a method for simultaneously realizing the utilization of the degradation products of the environment-friendly toilet in the well site and the solid waste of the water-based drilling.
In another aspect of the present invention, there is provided a method for cooperative biotreatment of water-based drilling solid waste by using wellsite environmental protection toilet degradants, the method comprising the steps of: uniformly mixing water-based drilling solid waste to be treated, inoculating 0.1-0.3% of microbial degradation strain with degradation effect according to the weight ratio, and fully and uniformly mixing; adding environment-friendly toilet degradation products with the solid waste amount of 0.5-1 wt% of the water-based drilling well to be treated, and fully and uniformly mixing; adding relatively fine fresh soil with the solid waste amount of 0.1-1 time of that of the to-be-treated soil, uniformly mixing and stacking; fresh soil with a certain thickness is covered on the surface of the treated object, ornamental or firewood plants are sown and planted on the fresh soil to form a well site solid waste-microorganism-plant combined degradation system.
In an exemplary embodiment of another aspect of the present invention, the method may further comprise the step of harmless treatment of the wellsite environmental toilet degradation product: degrading products of the well site environment-friendly toilet and edible fungus residues according to a weight ratio of 1: 2-3, uniformly mixing; adding 10-20 wt% of bran and 0.5-1 wt% of organic fertilizer into the degraded product of the well site environment-friendly toilet, and uniformly mixing; fermenting at 55-70 ℃ for 48-72 h, and stacking at normal temperature for 28-56 days.
In an exemplary embodiment of another aspect of the invention, the treated water-based drilling solid waste has a COD of 35mg/L, Oil of 0.8mg/L、Cr6+The content is less than or equal to 0.08mg/L, the ammonia nitrogen content is less than or equal to 1.6mg/L, the cadmium content is less than or equal to 2mg/kg, and the lead content is less than or equal to 530 mg/kg.
Compared with the prior art, the beneficial effects of the invention can comprise at least one of the following:
(1) the domestic solid waste generated by the drilling environment-friendly toilet is subjected to biodegradation treatment to obtain a degradation product, so that the transfer amount of the domestic solid waste in a well site is reduced;
(2) after harmless treatment is carried out on the degradation product, the degradation product is used as an additive or a nutrient to cooperate with biological treatment for resource utilization of water-based solid waste, so that the use of other biological nutrient additives in biological treatment is reduced;
(3) the drilling solid waste can be recycled by adopting the biological treatment on the spot, so that the solid waste disposal and utilization amount of the whole well transportation and the outward transportation to the local brick making factory can be greatly reduced, and finally, the solid waste transportation disposal amount, the disposal cost and the safety and environmental protection risk in the transportation can be reduced.
Drawings
The Bacillus belgii used in the invention is preserved, and the preservation unit is as follows: china Center for Type Culture Collection (CCTCC), preservation address: in the Wuhan university school of Wuhan 299 in the Wuchang area of Wuhan city, Hubei province, the preservation date is as follows: 29 months 5 in 2020, the preservation number is CCTCC NO: m2020160.
The above and other objects and features of the present invention will become more apparent from the following description taken in conjunction with the accompanying drawings, in which:
FIG. 1A shows the microscopic form of Bacillus belgii according to an exemplary embodiment of the present invention; FIG. 1B shows the colony morphology of Bacillus beleisi on beef extract peptone medium;
FIG. 2 shows a phylogenetic diagram of the 16S rDNA sequence of Bacillus belgii in an exemplary embodiment of the invention.
Detailed Description
Hereinafter, the method for preparing the wellsite eco-toilet degradant and the method for co-biologically treating water-based drilling solid waste of the wellsite eco-toilet degradant according to the present invention will be described in detail with reference to the exemplary embodiments and the accompanying drawings.
In an exemplary embodiment of the present invention, a method for preparing a degradation product of a wellsite environmental-friendly toilet comprises the steps of: after domestic wastewater and human excrement generated by the well site environment-friendly toilet are treated by a biological agent, filtering and drying are carried out until the water content is 30-55%, and the well site environment-friendly toilet degradation product is obtained. The biological agent is Bacillus velezensis B3-4 or a bacterial suspension thereof or a culture solution thereof or a fermentation product thereof, and the degradation product of the wellsite environment-friendly toilet can comprise 40-62% by weight of organic matters, 0.5-1.0% by weight of nitrogen, 1.2-1.5% by weight of phosphorus and 0.8-1.2% by weight of potassium. The degradation product of the well site environment-friendly toilet contains organic matters, nitrogen, phosphorus, potassium and other nutrient substances, is a high-quality raw material for organic fertilizer and soil fertilization, and can be used as an organic nutrient or additive for biological strain culture fermentation in biological treatment of water-based drilling solid waste.
In the present exemplary embodiment, the eco-friendly toilet may include a water-free eco-friendly toilet, and the bio-agent may be Bacillus beijerinckii (Bacillus velezensis) B3-4 or a bacterial suspension thereof or a culture solution thereof or a fermentation product thereof.
The Bacillus belgii is prepared by adopting a screening culture method. The strain is classified and named as Bacillus velezensis B3-4, is preserved in China Center for Type Culture Collection (CCTCC) in 5-29 th of 2020, and has the preservation number of CCTCC NO: m2020160. Bacillus velezensis B3-4 is a gram-positive bacterium, has spores, is rod-shaped, has motility and is obligately aerobic. Extracting total DNA of the strain B3-4, amplifying a 16S rDNA fragment, and determining a 16S sequence shown in SEQ ID NO. 1. The measured 16S sequence was aligned at the National Center for Biotechnology Information (NCBI) database of the United states, and B3-4 showed the highest similarity (99.6%) to the model strain Bacillus velezensis CR-502T in GenBank, thereby confirming that B3-4 is classified as Bacillus velezensis B3-4.
The bacterial colony formed after the Bacillus velezensis B3-4 is cultured on a beef extract peptone medium for 24 hours is round or irregular, and the bacterial colony after 48 hours is round, white (milky white), 0.5-1 mm in diameter, irregular in edge and flat and moist. In the embodiment, Bacillus velezensis B3-4 has stress resistance and can grow in a culture medium with a salt concentration of 0.5-4%.
The screening culture method of the Bacillus belgii comprises the following steps:
1) samples (e.g., human feces) are collected and cryopreserved, including human feces, livestock feces, soil, water, sludge, and organic fertilizers.
2) A predetermined amount (for example, 3 to 8g) of the dilution of the sample is applied to a beef extract peptone medium by a gradient dilution method, and incubated at a constant temperature of 15 to 50 ℃. Here, the sample is suitably used in an appropriate amount, and the temperature for cultivation may be adjusted depending on the temperature range of the environment to which the target strain is applied, for example, 5g of the sample may be used, 50 ℃ if the target strain is applied to a high temperature environment, 15 ℃ if the target strain is applied to a low temperature environment, and 30 ℃ if the target strain is applied to a normal temperature environment.
3) After the culture medium grows out of the bacteria, selecting bacterial colonies with different forms from a beef extract peptone flat plate for streak culture.
4) Subsequently (for example, after culturing for 16-18 h), a ring of colonies is picked for repeated streaking, and combined with microscopic observation until a purified strain is obtained, and the purified strain is inoculated into a test tube slant culture medium for preservation. The Bacillus velezensis B3-4 has strong degradation capability and can degrade and remove organic matters and COD (chemical oxygen demand) in human excrement and domestic wastewater.
Preferably, the bacterial suspension of Bacillus velezensis B3-4, or the culture solution thereof, or the fermentation product thereof also has strong degradation capability, and can effectively degrade and remove organic matters and COD in human excrement and domestic wastewater.
The Bacillus velezensis B3-4 or the biological agent has good degradation application potential and can be applied to biodegradable toilets.
In order that the above-described exemplary embodiments of the invention may be better understood, further description thereof with reference to specific examples is provided below.
Example 1
(1) Separation, purification and preservation of Bacillus velezensis B3-4 (referred to as "strain B3-4")
Collected human fecal samples are stored at low temperature and taken back to the laboratory. Taking 5g of a fecal sample, coating the fecal sample on a beef extract peptone culture medium by adopting a gradient dilution method, and culturing at constant temperature of 30 ℃. After the colonies grow out, colonies with different forms are selected from the plate and streaked on a beef extract peptone plate. And (5) after culturing for 16h, picking one ring for repeated streak culture, and combining microscopic observation until purification. And inoculating the purified strain into a test tube slant culture medium of a beef extract peptone culture medium for storage.
As shown in FIG. 1, the separated and purified strain B3-4 of the present example was cultured on a beef extract peptone medium, and the colony was circular, white, 0.5-1 mm in diameter, irregular in edge, and flat and wet after 48 hours of culture. Wherein FIG. 1A shows the colony morphology and FIG. 1B is a schematic representation of colonies after streaking on beef extract peptone plates.
(2) Amplification and phylogenetic analysis of 16S rDNA of strain B3-4
Extracting total DNA of the strain, using the total DNA as a template, using 27F and 1492R as primers to amplify a 16S fragment, and using a Bio-RADMCyclerTM instrument for PCR reaction.
Reaction system (50 μ l): mu.l of 2 XPCRMix 25. mu.l each of primers 27F and 1492R (10. mu.M), 1. mu.l of DNA template, and made up to 50. mu.l with ultrapure water; the nucleotide sequences of primers 27F and 1492R are shown in SEQ ID No.2 and SEQ ID No. 3.
And (3) PCR reaction conditions: pre-denaturation at 94 ℃ for 5 min; denaturation at 94 deg.C for 1min, annealing at 54 deg.C for 1min, extension at 72 deg.C for 2min, and circulation for 30 times; final extension at 72 ℃ for 8 min.
The PCR amplification product was detected on 1.0% agarose gel electrophoresis and then sent to Shanghai Bioengineering Co., Ltd for sequence determination. The software DNA man6.0 was used to perform the calculation of gene sequence similarity. The sequencing result is shown in SEQ ID No.1 (sequence table).
The obtained sequence results were compared in the National Center for Biotechnology Information (NCBI) database, and it was found that the 16S rDNA gene sequence of B3-4 had the highest similarity of 99.6% to B3-4 in the database and the model strain Bacillus velezensis CR-502T in GenBank. Based on the NCBI comparison results, the model strain with the highest similarity was selected as the reference strain, and phylogenetic trees (as shown in FIG. 2) and self-developed values (bootstraps) were constructed using the Neighbor-joining method of MEGA 6.0 software (Neighbor-joining).
Based on the above characteristics, the strain B3-4 was identified as Bacillus belgii (Bacillus velezensis). The strain is stored in China center for type culture Collection in 29 th 5 th 2020, and the preservation number is CCTCC NO: m2020160.
(3) Experiment on capability of bacterial strain B3-4 in degrading organic matters and COD in human excrement
(4) Resistance test of Strain B3-4
And (3) determining the salt tolerance of the strain B3-4: preparing a beef extract peptone culture medium, and adjusting the pH value to 7; adding NaC1 into basic beef extract peptone medium, and preparing culture medium with salt concentration of 0.5%, 1%, 2%, 4%, 8% and 10%. After sterilizing at 121 ℃ for 30min, adding the bacterial suspension to a newly configured culture medium according to 1% (v: v), repeating three times, placing the culture medium in a 30 ℃ constant temperature incubator, performing shake culture at the rotating speed of 150r/min for 48h, and measuring the number of bacteria by using an ultraviolet spectrophotometer under the wavelength lambda of 600.
Growth temperature Range determination of Strain B3-4: preparing a beef extract peptone culture medium, adjusting the pH value to 7, sterilizing at 121 ℃ for 30min, adding bacterial suspension to the newly-prepared culture medium according to 1% (v: v), placing the inoculated culture medium in a constant-temperature incubator with the temperature of 10 ℃, 20 ℃, 30 ℃, 40 ℃, 50 ℃ and 60 ℃ respectively, carrying out shake culture at the rotating speed of 150r/min for 48h, setting 3 times at each temperature, and measuring the quantity of bacteria by using an ultraviolet spectrophotometer under the wavelength lambda of 600.
The result shows that the Bacillus beleisi B3-4 has good stress resistance and stronger salt resistance, and can grow in a beef extract peptone culture medium containing 4% NaCl; the growth temperature range is wide, and the growth can be carried out within the temperature range of 15-50 ℃.
(5) Application of strain B3-4 in biodegradation type toilet experiment
Inoculating B3-4 strain in LB liquid culture medium, performing shaking culture at 28 deg.C, culturing for 24h, inoculating 5% into biodegradable ecological toilet with bamboo residue as filling matrix, adding appropriate amount of water, stirring, and adding their excrement into 2 students every day. Weekly faecal additions were recorded. The total weight was measured weekly. The experimental period was 17 weeks with no inoculation treatment as a control.
The dry matter reduction rate can be calculated by the following formula: r ═ S + F + S0)/(S + F) × 100%; in the formula, S is the mass of the added bamboo slag; f is the total dry matter amount of the added excrement; s0 is the dry matter quantity of the final mixture of the excrement and the bamboo slag.
The results show that: after the inoculation treatment is carried out for 17 weeks, 243.54kg of excrement is added, 231.35kg of excrement is reduced, the human excrement reduction rate is 95%, compared with the human excrement reduction rate of a control treatment, the human excrement reduction rate is increased by 87.5%, and good degradation application potential is shown.
In another exemplary embodiment of the invention, a method of cooperative biotreatment of water-based drilling solid waste with wellsite environmental toilet degradants comprises the steps of:
uniformly mixing water-based drilling solid waste to be treated, inoculating 0.1-0.3% of microbial degradation strain with degradation effect according to the weight ratio, and fully and uniformly mixing; adding environment-friendly toilet degradation products with the solid waste amount of 0.5-1 wt% of the water-based drilling well to be treated, and fully and uniformly mixing; adding relatively fine fresh soil with the solid waste amount of 0.1-1 time of that of the to-be-treated soil, uniformly mixing and stacking; fresh soil with a certain thickness is covered on the surface of the treated object, ornamental or firewood plants are sown and planted on the fresh soil to form a well site solid waste-microorganism-plant combined degradation system. The water-based drilling solid waste refers to solid waste generated by drilling by adopting a water-based drilling fluid system. The COD content in the treated water-based drilling solid waste is less than or equal to 35mg/L, Oil and less than or equal to 0.8mg/L, Cr6+The content is less than or equal to 0.08mg/L, the ammonia nitrogen content is less than or equal to 1.6mg/L, the cadmium content is less than or equal to 2mg/kg, and the lead content is less than or equal to 530 mg/kg. The microbial degradation strain can be one or a mixture of two of Halomonas ventosae and Halomonas campisalis in Halomonas according to any proportion. However, it is not limited toThe present invention is not limited thereto, and other microorganism-degrading bacteria having the same function may be used. The degradants may function as additives or organic nutrients. Specifically, the screened safe and advantageous liquid or solid microbial degradation strain is transported to a construction site, water-based drilling solid waste to be treated is fully mixed, the microbial degradation strain is directly added according to a certain proportion after the mixing is uniform, then the fine soil with 0.1-1 time of the solid waste amount to be treated is added according to the water content condition of the solid waste to be treated, the fine soil is stacked after the mixing is fully uniform, fresh soil is restored on the surface of the fine soil with the thickness of 2-5 cm, ornamental or firewood plants are sowed and planted on the fine soil to form a drilling solid waste-microorganism-plant combined degradation system, so that COD, Oil and Cr in the final product are enabled to be combined with the drilling solid waste-microorganism-plant combined degradation system, and then COD6+And the content of other heavy metals and the like reaches the standard of being used as the ploughing and planting soil.
In this embodiment, the method for the cooperative biological treatment of water-based drilling solid waste by the wellsite environment-friendly toilet degradation product can further comprise a harmless treatment step: the degradation product of the wellsite environment-friendly toilet prepared in the last exemplary embodiment and the edible fungus residues are mixed according to the weight ratio of 1: 2-3, and mixing uniformly. Adding 10-20 wt% of bran and 0.5-1 wt% of organic fertilizer into the degraded product of the well site environment-friendly toilet, and uniformly mixing; stacking and fermenting for 48-72 h at the temperature of 55-70 ℃, then stacking for 28-56 days at normal temperature, and removing harmful components in the degradation products of the well site environment-friendly toilet.
In summary, the beneficial effects of the invention can include at least one of the following:
(1) the degradation products of the drilling environment-friendly toilet are recycled;
(2) the water-based solid waste is recycled by cooperative biological treatment, so that the use of other biological nutrition additives in biological treatment is reduced;
(3) the drilling solid waste can be recycled by adopting the biological treatment on the spot, so that the solid waste disposal and utilization amount of the whole well transportation and the outward transportation to the local brick making factory can be greatly reduced, and finally, the solid waste transportation disposal amount, the disposal cost and the safety and environmental protection risk in the transportation can be reduced.
While the present invention has been described above in connection with exemplary embodiments, it will be apparent to those of ordinary skill in the art that various modifications may be made to the above-described embodiments without departing from the spirit and scope of the claims.
Sequence listing
<110> China Petroleum and gas group, Chuanqing drilling engineering, Inc., Sichuan Kort detection technology, Inc
<120> preparation method of degradation products of well site environment-friendly toilet and method for treating water-based solid waste
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1404
<212> DNA
<213> Bacillus belgii (Bacillus velezensis)
<400> 1
gctggcggcg tgcctaatac atgcaagtcg agcggacaga tgggagcttg ctccctgatg 60
ttagcggcgg acgggtgagt aacacgtggg taacctgcct gtaagactgg gataactccg 120
ggaaaccggg gctaataccg gatggttgtt tgaaccgcat ggttcagaca taaaaggtgg 180
cttcggctac cacttacaga tggacccgcg gcgcattagc tagttggtga ggtaacggct 240
caccaaggca acgatgcgta gccgacctga gagggtgatc ggccacactg ggactgagac 300
acggcccaga ctcctacggg aggcagcagt agggaatctt ccgcaatgga cgaaagtctg 360
acggagcaac gccgcgtgag tgatgaaggt tttcggatcg taaagctctg ttgttaggga 420
agaacaagtg ccgttcaaat agggcggcac cttgacggta cctaaccaga aagccacggc 480
taactacgtg ccagcagccg cggtaatacg taggtggcaa gcgttgtccg gaattattgg 540
gcgtaaaggg ctcgcaggcg gtttcttaag tctgatgtga aagcccccgg ctcaaccggg 600
gagggtcatt ggaaactggg gaacttgagt gcagaagagg agagtggaat tccacgtgta 660
gcggtgaaat gcgtagagat gtggaggaac accagtggcg aaggcgactc tctggtctgt 720
aactgacgct gaggagcgaa agcgtgggga gcgaacagga ttagataccc tggtagtcca 780
cgccgtaaac gatgagtgct aagtgttagg gggtttccgc cccttagtgc tgcagctaac 840
gcattaagca ctccgcctgg ggagtacggt cgcaagactg aaactcaaag gaattgacgg 900
gggcccgcac aagcggtgga gcatgtggtt taattcgaag caacgcgaag aaccttacca 960
ggtcttgaca tcctctgaca atcctagaga taggacgtcc ccttcggggg cagagtgaca 1020
ggtggtgcat ggttgtcgtc agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag 1080
cgcaaccctt gatcttagtt gccagcattc agttgggcac tctaaggtga ctgccggtga 1140
caaaccggag gaaggtgggg atgacgtcaa atcatcatgc cccttatgac ctgggctaca 1200
cacgtgctac aatggncaga acaaagggca gcgaaaccgc gaggttaagc caatcccaca 1260
aatctgttct cagttcggat cgcagtctgc aactcgactg cgtgaagctg gaatcgctag 1320
taatcgcgga tcagcatgcc gcggtgaata cgttcccggg ccttgtacac accgcccgtc 1380
acaccacgag agtttgtaac accc 1404
<210> 2
<211> 19
<212> DNA
<213> Artificial sequence (27F)
<400> 2
agagttgatc ctggctcag 19
<210> 3
<211> 20
<212> DNA
<213> Artificial sequence (1492R)
<400> 3
cggttacctt gttacgactt 20
Claims (9)
1. A preparation method of a degradation product of a wellsite environment-friendly toilet is characterized by comprising the following steps:
treating domestic wastewater and human excrement generated by the well site environment-friendly toilet by using a biological agent, and filtering and drying the treated domestic wastewater and human excrement until the water content is 30-55% to obtain a well site environment-friendly toilet degradation product;
wherein the biological agent is Bacillus belgii B3-4 or its suspension or its culture solution or its fermentation product.
2. The method for preparing the degradation product of the wellsite environmental-friendly toilet according to claim 1, wherein the environmental-friendly toilet is a water-free environmental-friendly toilet.
3. The method for preparing degradation products in environmental-friendly toilets at wellsites as claimed in claim 1, wherein Bacillus belgii is gram-positive, spore-like, rod-like, motile, and obligatory aerobic.
4. The method for preparing the degradation product of the wellsite environment-friendly toilet of claim 1, wherein the 16S rDNA sequence of the Bacillus belgii is shown in SEQ ID No. 1.
5. The preparation method of the degradation product of the wellsite environment-friendly toilet as claimed in claim 1, wherein bacterial colonies formed by the Bacillus belgii after being cultured on a beef extract peptone medium for 24 hours are round or irregular, and bacterial colonies after 48 hours are round, white, 0.5-1 mm in diameter, irregular in edge and flat and wet.
6. The method for preparing the degradation products in the wellsite environment-friendly toilet as claimed in claim 1, wherein the screening culture method of Bacillus belgii comprises the following steps:
collecting and storing samples at low temperature, wherein the samples comprise human excrement, livestock and poultry excrement, soil, water, sludge and organic fertilizer;
coating the diluent of the sample on a beef extract peptone culture medium by adopting a gradient dilution method, and culturing at a constant temperature of 15-50 ℃;
after the culture medium grows out of the bacteria, selecting bacterial colonies with different forms from a beef extract peptone flat plate for streak culture;
subsequently, a loop of colonies was picked for repeated streaking with microscopic observation until a purified strain was obtained.
7. A method for cooperative biological treatment of water-based drilling solid waste with wellsite environmental-friendly toilet degradants, the method comprising the steps of:
uniformly mixing water-based drilling solid waste to be treated, inoculating 0.1-0.3% of microbial degradation strain with degradation effect according to the weight ratio, and fully and uniformly mixing;
adding environment-friendly toilet degradation products with the solid waste amount of 0.5-1 wt% of the water-based drilling well to be treated, and fully and uniformly mixing;
adding relatively fine fresh soil with the solid waste amount of 0.1-1 time of that of the to-be-treated soil, uniformly mixing and stacking;
fresh soil with a certain thickness is covered on the surface of the treated object, ornamental or firewood plants are sown and planted on the fresh soil to form a well site solid waste-microorganism-plant combined degradation system.
8. The method of cooperative biotreatment of water based drilling solid waste using wellsite environmental friendly toilet degradants as claimed in claim 7, further comprising the step of subjecting the wellsite environmental friendly toilet degradants to a harmless treatment:
degrading products of the well site environment-friendly toilet and edible fungus residues according to a weight ratio of 1: 2-3, uniformly mixing;
adding 10-20 wt% of bran and 0.5-1 wt% of organic fertilizer into the degraded product of the well site environment-friendly toilet, and uniformly mixing;
fermenting at 55-70 ℃ for 48-72 h, and stacking at normal temperature for 28-56 days.
9. The method of claim 7, wherein the treated water-based drilling solid waste has a COD of 35mg/L, Oil content of 0.8mg/L, Cr6+The content is less than or equal to 0.08mg/L, the ammonia nitrogen content is less than or equal to 1.6mg/L, the cadmium content is less than or equal to 2mg/kg, and the lead content is less than or equal to 530 mg/kg.
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