CN114058529B - Oily pollutant degradation strain ZT2, microbial agent and application - Google Patents
Oily pollutant degradation strain ZT2, microbial agent and application Download PDFInfo
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- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F11/00—Treatment of sludge; Devices therefor
- C02F11/02—Biological treatment
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- 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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Abstract
The invention provides an oily pollutant degradation strain ZT2, a microbial agent and application. The pollutant-containing degradation strain is Bacillus tervelopneum (Bacillus tequilensis) ZT2, the preservation number of the strain is CGMCC No.17158, and the invention can achieve the effect of greatly reducing the petroleum hydrocarbon content of the sludge by degrading the thick oil sludge through the single strain of the Bacillus tervelopneum (Bacillus tequilensis) ZT2. In the indoor test, the degradation rate of the total petroleum hydrocarbon content in the thick oil sludge inoculated with the bacillus tervelarius (Bacillus tequilensis) strain for 21 days is 17.69%. In an outdoor test, the degradation rate of the total petroleum hydrocarbon content in the thick oil sludge inoculated with the Bacillus tequila (Bacillus tequilensis) strain for 52 days is 50.59 percent.
Description
Technical Field
The invention relates to the technical field of microbial remediation, in particular to an oily pollutant degradation strain ZT2, a microbial agent and application.
Background
The oily sludge is listed in the national hazardous waste list in China, and the national requirements of the oily sludge are that the oily sludge must be subjected to harmless treatment to realize recycling. The capacity of reducing treatment and recycling of the oily sludge not only restricts the survival and development of enterprises, but also has great hidden environmental pollution. At present, the treatment of the oily sludge mainly adopts a chemical heat washing method, an extraction separation method, a biological treatment method, an incineration method and the like, but the traditional chemical heat washing method is not clean and thorough for the oily sludge with relatively complex components, especially for the thick oil sludge, and has poor reduction effect; the extraction separation method has high treatment cost and low recovery and reuse efficiency of the extractant; the incineration method has higher cost, serious secondary pollution and larger one-time investment. At present, the treatment methods such as chemical heat washing, extraction and the like have certain limitations, and the microbial degradation of the oily sludge has the advantages of high safety, no secondary pollution, low cost, environmental friendliness and the like, and is considered as an economic and environmental-friendly method for treating the oily sludge, so how to screen and obtain the efficient petroleum degradation strain is a key for treating the oily sludge by using a biological method.
Chinese patent application 201810115869.9 discloses a strain of efficient degradation bacteria JN6 of petroleum hydrocarbons in oily sludge and application thereof, wherein the strain is Comamonas sp. The strain JN6 with petroleum hydrocarbon degradation function is used for treating the oily sludge in a petroleum hydrocarbon degradation mode, the degradation rate of the strain on Total Petroleum Hydrocarbon (TPH) in the oily sludge within 30 days is 62.16%, and the optimal degradation condition of the strain on petroleum pollutants is 15-40 ℃ and the pH value is 7-11. Chinese patent application 201810115860.8 discloses a strain of efficient degradation bacteria JN4 for petroleum hydrocarbon in oily sludge and application thereof, wherein the strain is Pseudomonas stutzeri Pseudomonas stutzeri. The strain JN4 with the petroleum hydrocarbon degradation function is used for treating the oil-containing sludge in a petroleum hydrocarbon degradation mode. The bacterium has a degradation rate of 71.05% for Total Petroleum Hydrocarbon (TPH) in oil-containing sludge within 30 days, and has obvious degradation capability for n-C10-n-C30, and GC-MS analysis shows that the bacterium has strong degradation capability for naphthene decalin, indene and homoandrostane, and the optimal degradation condition for petroleum pollutants is 25-40 ℃ and pH value is 5-11. Chinese patent application 201710022605.4 discloses a Klebsiella (Klebsiella sp.) bacterium CGMCC13201 which can effectively degrade crude oil in crude oil contaminated soil such as oil-containing sludge, oil-containing silt and the like. The invention discloses a Chinese patent application 201410072228.1, which relates to a petroleum degrading bacterium in oily sludge, wherein the strain is corynebacterium glutamicum separated from the oily sludge of an oil field of Bohai sea. The preservation number of the bacterium CGMCC8647 proves that the degradation rate of RS1 to saturated hydrocarbon is higher, reaching 20.74 percent, the degradation rate of the bacterium 30d to Total Petroleum Hydrocarbon (TPH) in oil-containing sludge is 39.69 percent, and the optimal degradation condition of the bacterium to petroleum is 37 ℃, the salinity is 0.5-5 percent and the pH is 6-8.
However, the degradation bacteria mentioned above have limited degradation range for oily contaminants, and do not perform well for oily sludge, especially residual thick oil sludge after treatment by thick oil sludge chemical fine heat washing treatment technology, so that it is highly demanded to find a microbial strain for degrading oily contaminants better.
Disclosure of Invention
In order to improve the limitation of the existing microorganisms on the degradation range of oil-containing pollutants, the invention aims to provide the microbial Bacillus tequilensis ZT2 for degrading the oil-containing pollutants, which can effectively degrade the residual thick oil sludge after the treatment of the thick oil sludge by a chemical fine heat washing treatment technology.
Another object of the present invention is to provide a microbial agent containing the above bacillus tertiaryii ZT2.
It is a further object of the present invention to provide the use of the microorganism as described above and microbial agents thereof for the degradation and remediation of oily contaminants.
The aim of the invention is achieved by the following technical scheme:
a bacillus tertiarygensis (Bacillus tequilensis) ZT2, the bacillus tertiarygensis ZT2 having been deposited in a preservation unit under the preservation unit name: china general microbiological culture Collection center (CGMCC) with the address: the institute of microorganisms of national academy of sciences of China, national institute of sciences, no.1, no. 3, north Chen West Lu, the Korean region of Beijing; preservation date: the collection number of the strain is CGMCC NO.17158 and is classified and named Bacillus tequilensis on the 1 st and 11 th 2019.
According to some embodiments of the invention, the nucleotide sequence of the 16S rDNA of the bacillus tertiarygensis ZT2 is shown in SEQ ID NO. 1.
The colony morphology of the bacillus tertiaryalis ZT2 is as follows: light white, round, translucent, smooth, moist, clean-edged, gram-negative bacilli. According to some embodiments of the present invention, the degradation condition of the bacillus tertiaryii ZT2 on petroleum hydrocarbons is 10-30 ℃, and the pH value is 7-8, preferably pH value 8.
On the other hand, the invention also provides a microbial agent, which contains the bacillus tervelarii ZT2, and can be added with other conventional auxiliary components such as talcum powder, tween and the like, and the preparation method of the microbial agent can be obtained through a conventional test.
In still another aspect, the invention also provides an application of the bacillus terveleaf ZT2 in degrading and repairing oily pollutants.
According to some embodiments of the invention, the oil-containing contaminants include oil-containing sludge, water bodies contaminated with crude oil, and crude oil contaminants.
According to some embodiments of the invention, the oil-containing sludge comprises thickened oil sludge which is remained after treatment by a thickened oil sludge chemical fine heat washing treatment technology, preferably thickened oil sludge with low oil content.
According to some embodiments of the invention, the above application comprises the steps of: inoculating bacillus tervelarium ZT2 into a culture medium for culture to prepare a bacterial suspension; and step two, mixing the bacterial suspension with the oil-containing pollutant, repeatedly inoculating after a proper time, and then preserving for a proper time to finish degradation and repair.
According to a specific embodiment of the present invention, in step one, the medium is PDA (potato dextrose agar) medium without agar added.
According to a specific embodiment of the invention, in the first step, the condition that the bacillus tertiaryalis ZT2 is inoculated into a culture medium for culture is 30 ℃,180 r/min and shake is carried out for 3 days.
According to a specific embodiment of the invention, in step two, the inoculation is repeated after mixing the bacterial suspension with the oily contaminants for 13-15 days.
According to some embodiments of the invention, the PDA medium without agar addition is prepared from the following materials: each 1000mL of water contains 200g of potato and 20g of glucose.
According to some embodiments of the invention, the oleaginous contaminant is from a LiaoHe field.
According to some embodiments of the invention, the pH of the oleaginous contaminant is from 8 to 9 (soil: water=1:5).
According to some embodiments of the invention, the above application comprises the steps of: step one, inoculating bacillus tervelarii ZT2 into a PDA culture medium (without agar), culturing, and oscillating at 30 ℃ for 3 days at 180r/min to prepare a bacterial suspension; collecting the same batch of oily sludge in a container as a treatment, adding 300ml of the bacterial suspension into the container, stirring, repeating inoculation after 2 weeks, and setting a control treatment without adding a bacterial agent; step three, stirring (manual stirring) is performed every two weeks during the test to provide oxygen required for microbial degradation of petroleum. In the experimental process, part of water is properly added according to the dry and wet conditions of the crude oil containing the silt, so that the crude oil containing the silt is in a wet state in the experimental process, a plastic box is covered by a weather such as heavy rain, the total petroleum hydrocarbon content and the water content of the soil are measured once every half month in the first month of the test, and the total petroleum hydrocarbon content and the water content of the soil are measured once every month from the second month, and the statistical result is obtained.
According to some embodiments of the invention, the volume of the vessel in which the oleaginous contaminants are collected is 40cm by 55cm by 6cm.
The invention further provides application of the bacillus tertiarygensis ZT2 in degrading and repairing oily pollutants.
The invention has the beneficial effects that:
1. the bacillus tertiaryalis (Bacillus tequilensis) ZT2 is derived from local oily sludge of Liaohe oil fields, and the strain enters into a soil flora to occupy a certain ecological niche, so that the degradation of petroleum hydrocarbons is effectively promoted.
2. According to the invention, the single strain of Bacillus tequila (Bacillus tequilensis) ZT2 is used for degrading the thickened oil sludge, so that the thickened oil sludge reduction effect can be achieved. In the indoor test, the degradation rate of the total petroleum hydrocarbon content in the thick oil sludge inoculated with the bacillus tervelarius (Bacillus tequilensis) strain for 21 days is 17.69%. In an outdoor test, the degradation rate of the total petroleum hydrocarbon content in the thick oil sludge inoculated with the Bacillus tequila (Bacillus tequilensis) strain for 52 days is 50.59 percent. The strain can occupy a certain ecological place in the indigenous strain, promote the degradation of petroleum hydrocarbon, further reduce the total petroleum hydrocarbon content in the sludge treated by the thick oil fine hot washing thick oil sludge chemical fine hot washing treatment technology, and has lower treatment cost, good application effect and obvious economic and social benefits.
3. The Bacillus tertefrasis (Bacillus tequilensis) ZT2 realizes the excellent degradation of petroleum hydrocarbon in the oily sludge by a single strain, reduces the complex steps of activating each strain respectively and mixing the strains in proportion in the preparation process of the mixed microbial inoculum, is easy to control the optimal reaction condition for degrading the petroleum hydrocarbon in the oily sludge, and is easy to further remove the petroleum hydrocarbon in the oily sludge by combining other methods.
Drawings
FIG. 1 shows the growth of the culture medium of the strain ZT2 acclimatized in example 1 of the present invention.
FIG. 2 shows the ZT2 form of Bacillus teryaki (Bacillus tequilensis) obtained in example 1 of the present invention.
FIG. 3 is an electron microscope image of Bacillus tervelopneum (Bacillus tequilensis) ZT2 of example 1 of the present invention.
FIG. 4 shows degradation curves of Bacillus tertefragensis (Bacillus tequilensis) ZT2 in example 3 of the present invention for petroleum hydrocarbons in oily sludge at different pH values.
FIG. 5 shows degradation curves of Bacillus tertefragensis (Bacillus tequilensis) ZT2 in example 4 of the present invention for petroleum hydrocarbons in oily sludge at different temperatures.
The information on the preservation of the bacillus tertiaryalis ZT2 microorganisms is as follows:
preservation date: 2019, 1 month and 11 days;
preservation unit name: china general microbiological culture Collection center (CGMCC);
deposit unit address: the institute of microorganisms of national academy of sciences of China, national institute of sciences, no.1, no. 3, north Chen West Lu, the Korean region of Beijing;
preservation number: CGMCC No.17158;
classification naming: bacillus tequilensis Bacillus tequilensis.
Detailed Description
The following detailed description of the present invention will be presented in order to provide a clearer understanding of the technical features, objects and advantages of the present invention, but should not be construed as limiting the scope of the invention.
Example 1 screening and identification of Bacillus tertagatous ZT2
Material preparation
1 soil material
Taking petroleum polluted soil from a place of Liaohe oil field, carrying out scattering treatment, and storing at 4 ℃.
2 culture medium:
inorganic salt enrichment medium (1L): k (K) 2 HPO 4 ·3H 2 O 1g，KH 2 PO 4 1g，Mg 2 SO 4 ·7H 2 O 0.5g，NH 4 NO 3 1g，CaCl 2 0.02g，FeCl 3 Trace amount, diesel 3, pH 7.2-7.4.
PDA culture medium (1L) potato 200g; glucose 20g; 18-20 g of agar; 1000ml of distilled water; natural pH.
Degradation medium (1L): (NH 4) 2 SO 4 0.5g，NaNO 3 0.5g，CaCl 2 0.02g，MgSO 4 0.2g，KH 2 PO 4 1.0g，NaH 2 PO 4 ·H 2 O1.0 g, pH 7, and 50g of crude oil containing silt.
Isolation, screening and domestication of strains (II)
Adding 1% (w/v) petroleum oil polluted soil or residual oil into an inorganic salt enrichment culture medium taking diesel oil as a unique carbon source, carrying out shake cultivation for 3d at 30 ℃, inoculating 1mL of enrichment liquid into the same fresh culture medium, and carrying out cultivation for 3d under the same condition, thus continuously carrying out enrichment cultivation for 4 times. After the second culture, a large amount of microorganisms were observed to be produced in the culture medium, and the culture medium was in a suspended state.
After gradient dilution of the enrichment medium, the enrichment medium is respectively subjected to plate culture in PDA culture medium for 3 days, and a large number of microbial colonies are generated on the surface of the culture medium. After the bacterial colony grows out of the flat plate, a proper amount of single bacterial colony with different colors and morphologies is picked by an inoculating loop, and repeated streaking, separation and purification are carried out until the single bacterial colony is obtained. Uniformly numbering, and preserving at 4 ℃ inclined plane.
The oil sludge sample for domestication test is the oil sludge sample to be treated, and the oil content is 20%. And (5) carrying out scattering treatment on the soil sample, and sealing at 4 ℃ for later use. The strain obtained by separation and purification was inoculated with an inoculating needle into a sterilized 500ml Erlenmeyer flask containing 200ml of a liquid medium for degradation, and cultured at 30℃for 14 days by shaking at 180 rpm. After the completion of the culture, 0.1ml of each culture was cultured in a plate medium, and the growth was observed. The strain domesticated and cultured by the high oil-containing culture medium grows a large amount of strains on the PDA culture medium (figure 1), which shows that the strain screened by enrichment and separation can grow in the culture medium taking high-concentration petroleum as a carbon source.
DNA identification and morphological identification were performed by the Qingdao West letter detection analysis Co., ltd. The colony morphology of the strain ZT2 is as follows: as shown in FIG. 2, there are graininess, irregular wetting, flattening, uneven edges, and gram positive bacilli. The bacterial morphology is shown in scanning electron microscopy figure 3.
The sequence of the 16S ribosomal RNA gene of the strain is compared with a database by an analysis method for analysis, and the strain is found to belong to Bacillus tequila (Bacillus tequilensis), and the DNA sequence table of the strain is shown in the sequence table.
Example 2 degradation repair test of certain oily sludge in Liaohe oilfield
The thick oil sludge sample for degradation test is collected in the mud sand crude oil in the treatment process of a certain treatment plant in the period of 5.14.2018, and the soil is air-dried, scattered, ground and sealed for later use. Degradation test the strain ZT2 obtained by separation and purification was inoculated with an inoculating needle into a sterilized 250ml triangular flask containing 100ml of a liquid medium for degradation, and a medium not inoculated with a microorganism was set as a control group (CK), and the culture was shake-cultured at 30℃and 180rpm for 21 days, and each treatment was repeated three times. And after drying, measuring the total petroleum hydrocarbon content in the oily sludge by using an infrared spectroscope. The bacterium has good degradation effect on thick oil sludge, after 21 days of culture, the total petroleum hydrocarbon content of CK group is 5.40+/-0.31%, the total petroleum hydrocarbon content in thick oil sludge inoculated with ZT2 strain is 4.45+/-0.04%, and the degradation rate is 17.69% in 21 days.
Example 3 degradation effects of Bacillus tertefraensis (Bacillus tequilensis) on Petroleum hydrocarbons in oil sludge at different pH values
And collecting thick oil sludge in the treatment process of a certain treatment plant in 7.23.2018, air-drying, scattering and grinding the soil, and sealing the bag for later use. 100ml of degradation culture medium is added into a 250ml conical flask, the initial pH value of the degradation culture medium for degrading liquid inorganic salt is adjusted to be 5.0, 6.0, 7.0, 8.0 and 9.0 respectively by NaOH or HCl, the strain ZT2 obtained in the separation and purification in the example 1 is inoculated by an inoculating needle after sterilization, the culture is carried out for 14 days by shaking culture by a constant temperature shaking table at 30 ℃ and 180rpm, the total petroleum hydrocarbon content in the oily sludge is measured by an infrared spectroscope after drying, and the degradation rate is calculated. The degradation effect of the strain Bacillus tequilensis (Bacillus tequilensis) on petroleum hydrocarbons in oily sludge under different pH conditions is shown in figure 4.
The oil content of the control treated (CK) soil sample without inoculating microorganisms is 7.19+/-0.36%, the petroleum degradation effect is good in the test range of pH 7-8, the degradation rate is 15.11% -16.73%, and the degradation effect is optimal under the condition of pH 8.
Example 4 degradation effects of Bacillus tertefraensis (Bacillus tequilensis) on Petroleum hydrocarbons in oil sludge at different temperatures
And collecting thick oil sludge in the treatment process of a certain treatment plant in 7.23.2018, air-drying, scattering and grinding soil, and bagging and sealing for later use. Adding 100ml of degradation culture medium into a 250ml conical flask, regulating the initial pH of the degradation culture medium for degrading the liquid inorganic salt to 7.0 by using NaOH or HCl, inoculating the strain ZT2 obtained by separation and purification in the example 1 by using an inoculating needle after sterilization, respectively setting the temperature of a shaking table to 10 ℃, 20 ℃,30 ℃, 40 ℃ and 180rpm for shaking culture for 14 days, drying, measuring the total petroleum hydrocarbon content in the oily sludge by using an infrared spectroscope, and calculating the degradation rate. The degradation effect of the strain of Bacillus tequilensis (Bacillus tequilensis) on petroleum hydrocarbons in oily sludge at different temperatures is shown in FIG. 5. The oil content in the control treated (CK) soil sample without inoculating microorganism is 7.24+ -0.14%, the optimal culture temperature of ZT2 is 10-30deg.C, and the degradation rate is 12.69-19.73%.
EXAMPLE 5 Bacillus tertakii (Bacillus tequilensis) remediation of actual oily sludge
Based on the above examples, bacillus tertiaryother (Bacillus tequilensis) was used to repair actual oily sludge, and the experimental procedures were as follows:
(1) Test oily sludge: in 2018, 8 and 3 days, in a field for recycling crude oil containing silt in Liaohe oil field, placing a batch of treated thick oil sludge into the same plastic box with the volume of 40cm multiplied by 55cm multiplied by 6cm, adding microbial agent into the plastic box, and setting control treatment without microbial agent. The pH of the test sludge sample was 8.4 (soil: water=1:5).
(2) Preparation of bacterial suspension of the strain Bacillus tequilensis (Bacillus tequilensis):
bacillus tertequila (Bacillus tequilensis) ZT2 was selected and inoculated into PDA medium (without agar) for culture, and shaking at 30deg.C and 180r/min for 3 days for the test. 300ml of bacterial liquid is added in each treatment, and stirring is carried out. Repeated inoculations were performed after 2 weeks.
Stirring (with manual stirring) was performed every two weeks during the test to provide the oxygen required for microbial degradation of petroleum. In the experimental process, part of water is properly added according to the dry and wet conditions of the crude oil containing the silt, so that the crude oil containing the silt is in a wet state in the experimental process, and the upper cover of the plastic box is covered in the weather such as heavy rain. The total petroleum hydrocarbon content of the soil is measured once every half month in the first month of the test (the total petroleum hydrocarbon content of the soil is actually the oil content, because the collected samples are wet samples, the directly measured values are greatly influenced by the water content of the soil, the dry oil content needs to be calculated for comparison of the samples), and the water content is measured once a month from the second month.
When the first sampling is carried out in 8 months and 3 days, 3 groups of crude oil samples containing silt are taken, the oil content (dry) =total petroleum hydrocarbon content of soil/(1-water content) ×100%) of the crude oil samples is between 2.18 and 2.96%, but the error between the oil contents of the thick oil sludge on the surface layer and the thick oil sludge on the bottom layer is larger when the sampling is found in the test. Thus, the oil content of each group on 8 months and 17 days was used as the initial value of each group of strains, and the degradation rate after the addition of petroleum for 52 days (10 months and 8 days) was calculated, and the results are shown in Table 1:
TABLE 1
After 52 days, the oil content of the control group is reduced from 2.05% to 1.38%, and the degradation rate reaches 32.35%. The soil bacteria has a certain degradation effect. After the Bacillus tertequila (Bacillus tequilensis) ZT2 is added, the thick oil sludge with the oil content of 2.28 percent is degraded to 1.13 percent, and the degradation rate reaches 50.59 percent. The degradation function bacteria are added to promote the degradation of the thick oil sludge. The pH of the sample of the thick oil sludge to be tested is 8.4 (soil: water=1:5), and the degradation speed is high at 10-30 ℃, which accords with the optimal degradation condition that the degradation temperature is 10-30 ℃ and the pH is 7-8.
Sequence listing
<110> China Petroleum and Natural gas stock Co., ltd
<120> an oily pollutant degrading strain ZT2, microbial agent and application
<130> GAI20CN2952
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1382
<212> DNA
<213> Bacillus tertefraxins (Bacillus tequilensis)
<400> 1
gatgggagct tgctccctga tgttagcggc ggacgggtga gtaacacgtg ggtaacctgc 60
ctgtaagact gggataactc cgggaaaccg gggctaatac cggatggttg tttgaaccgc 120
atggttcaaa cataaaaggt ggcttcggct accacttaca gatggacccg cggcgcatta 180
gctagttggt gaggtaacgg ctcaccaagg caacgatgcg tagccgacct gagagggtga 240
tcggccacac tgggactgag acacggccca gactcctacg ggaggcagca gtagggaatc 300
ttccgcaatg gacgaaagtc tgacggagca acgccgcgtg agtgatgaag gttttcggat 360
cgtaaagctc tgttgttagg gaagaacaag taccgttcga atagggcggt accttgacgg 420
tacctaacca gaaagccacg gctaactacg tgccagcagc cgcggtaata cgtaggtggc 480
aagcgttgtc cggaattatt gggcgtaaag ggctcgcagg cggtttctta agtctgatgt 540
gaaagccccc ggctcaaccg gggagggtca ttggaaactg gggaacttga gtgcagaaga 600
ggagagtgga attccacgtg tagcggtgaa atgcgtagag atgtggagga acaccagtgg 660
cgaaggcgac tctctggtct gtaactgacg ctgaggagcg aaagcgtggg gagcgaacag 720
gattagatac cctggtagtc cacgccgtaa acgatgagtg ctaagtgtta gggggtttcc 780
gccccttagt gctgcagcta acgcattaag cactccgcct ggggagtacg gtcgcaagac 840
tgaaactcaa aggaattgac gggggcccgc acaagcggtg gagcatgtgg tttaattcga 900
agcaacgcga agaaccttac caggtcttga catcctctga caatcctaga gataggacgt 960
ccccttcggg ggcagagtga caggtggtgc atggttgtcg tcagctcgtg tcgtgagatg 1020
ttgggttaag tcccgcaacg agcgcaaccc ttgatcttag ttgccagcat tcagttgggc 1080
actctaaggt gactgccggt gacaaaccgg aggaaggtgg ggatgacgtc aaatcatcat 1140
gccccttatg acctgggcta cacacgtgct acaatggaca gaacaaaggg cagcgaaacc 1200
gcgaggttaa gccaatccca caaatctgtt ctcagttcgg atcgcagtct gcaactcgac 1260
tgcgtgaagc tggaatcgct agtaatcgcg gatcagcatg ccgcggtgaa tacgttcccg 1320
ggccttgtac acaccgcccg tcacaccacg agagtttgta acacccgaag tcggtgaggt 1380
aa 1382
Claims (12)
1. Bacillus tertequilensisBacillus tequilensis) ZT2, characterized in that it has the strain deposit number: CGMCC No.17158.
2. A microbial agent comprising bacillus tertiarygensis ZT2 according to claim 1.
3. Use of bacillus tertiaryii ZT2 according to claim 1 for degradation remediation of oily contaminants.
4. Use according to claim 3, wherein the oleaginous contaminants comprise oleaginous sludge or bodies of water contaminated with crude oil.
5. The use according to claim 3, characterized by the following steps:
inoculating bacillus tervelarium ZT2 into a culture medium for culture to prepare a bacterial suspension;
and step two, mixing the bacterial suspension with the oil-containing pollutant, repeatedly inoculating after a proper time, and then preserving for a proper time to finish degradation and repair.
6. The method according to claim 5, wherein in step one, the medium is PDA medium without agar.
7. The use according to claim 5, wherein in step one, the bacillus tertefraxinus ZT2 is inoculated into a culture medium for culturing at 30 ℃ and 180r/min, and is shaken for 3 days.
8. The method according to claim 5, wherein in step two, the inoculation is repeated after mixing the bacterial suspension with the oily contaminants for 13-15 days.
9. The use according to claim 6, wherein the PDA medium without agar is formulated from the following materials: each 1000mL of water contains 200g of potato and 20g of glucose.
10. The use of claim 5, wherein the oleaginous contaminant is from a Liaohe field.
11. The use according to claim 5, wherein the pH of the oleaginous contaminant is from 8 to 9.
12. Use of the microbial agent of claim 2 for degrading and repairing oily contaminants.
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