CN111440756A - Method for treating petroleum sewage by using thermophilic bacteria - Google Patents
Method for treating petroleum sewage by using thermophilic bacteria Download PDFInfo
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- CN111440756A CN111440756A CN202010443076.7A CN202010443076A CN111440756A CN 111440756 A CN111440756 A CN 111440756A CN 202010443076 A CN202010443076 A CN 202010443076A CN 111440756 A CN111440756 A CN 111440756A
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Images
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/36—Adaptation or attenuation of cells
-
- 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
- C02F3/343—Biological treatment of water, waste water, or sewage characterised by the microorganisms used for digestion of grease, fat, oil
-
- 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
- C02F3/344—Biological treatment of water, waste water, or sewage characterised by the microorganisms used for digestion of mineral oil
-
- 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
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2101/00—Nature of the contaminant
- C02F2101/30—Organic compounds
- C02F2101/32—Hydrocarbons, e.g. oil
-
- 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
- C02F2103/10—Nature of the water, waste water, sewage or sludge to be treated from quarries or from mining activities
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W10/00—Technologies for wastewater treatment
- Y02W10/10—Biological treatment of water, waste water, or sewage
Abstract
The invention relates to the technical field of petroleum sewage treatment by a biological method, and particularly discloses a method for treating petroleum sewage by using thermophilic bacteria. The invention realizes the high-efficiency treatment of the single-strain thermophagia on the high-temperature petroleum sewage by the training of the thermophagia and the preparation of the thermophagia filler.
Description
Technical Field
The invention relates to the technical field of petroleum sewage treatment by a biological method, in particular to a method for treating petroleum sewage by using thermophilic bacteria.
Background
Oil, known as "industrial blood", is stored in the upper portion of the crust. The main component is a mixture of various alkanes, cycloalkanes and aromatics. With the development of economy, the demand of petroleum is greatly increased, however, petroleum sewage generated in the petroleum exploitation process becomes an important source of environmental pollution, and therefore, how to effectively treat the petroleum sewage is a common problem for researchers in various fields. In 2017, 10 and 27, the national health organization international cancer research institution publishes a carcinogen list for preliminary reference arrangement, and petroleum is refined in a 2A class carcinogen list. Therefore, when petroleum becomes an environmental pollution, it poses a very serious threat to human health.
The environmental problem of biological treatment is an eco-friendly priority, most of the petroleum sewage treatment at home and abroad at present only meets the requirement under normal temperature environment, the high-temperature petroleum sewage is one of the important sources of the petroleum sewage, the treatment difficulty is high, the record of the high-temperature petroleum treatment by using microorganisms in the prior art is very few, the microbial treatment only relates to the treatment of composite strains, the degradation rate of the high-temperature petroleum is below 65%, and the degradation rate of a single strain for treating the high-temperature petroleum is not up to 65% at present.
Disclosure of Invention
In order to solve the technical problems, the invention provides a method for treating petroleum sewage by using thermophilic bacteria, which comprises the following steps:
s1, activating thermoacidophilic bacteria, training the activated strains, and then culturing the trained strains at 70-80 ℃ for 22-26h to obtain bacterial liquid;
s2, loading the bacterial liquid obtained in the step S1 on a porous active carrier to obtain thermophagia bacteria filler, and then putting the thermophagia bacteria filler into petroleum wastewater for treatment, wherein the temperature of the petroleum wastewater is 50-80 ℃;
preferably, in S1, the culture medium for culturing Thermus thermophilus is culture medium I, and the formula of the culture medium I is 0.5 g/L NH4NO38 g/L tryptone, 4 g/L yeast extract, 3 g/L NaCl and 1L distilled water, and the pH of the medium I was 7.0 to 7.5.
Preferably, in S1, the training includes the following stages:
in the first training stage, the Thermus thermophilus is inoculated in a second culture medium and cultured for 20-28h at the temperature of 120-55 ℃ and at the rpm/min of 160-;
in the second training stage, the Thermus thermophilus which passes through the first training stage is inoculated in a second culture medium according to the inoculation amount of 8-18 percent, and cultured for 20-28h at the temperature of 60-65 ℃ and at the rpm of 120-;
in the third training stage, the thermus thermophilus passing through the second training stage is inoculated in a second culture medium according to the inoculation amount of 8-18 percent, and cultured for 20-28h at the temperature of 70-75 ℃ and at the rpm of 120-;
in the fourth training stage, the thermus thermophilus which passes through the second training stage is inoculated in a second culture medium according to the inoculation amount of 8-18 percent, and the culture is carried out for 20-28h at the temperature of 80 ℃ and at the rpm of 120-;
the second culture medium is an inorganic salt culture medium which takes petroleum as a unique carbon source.
Preferably, the formula of the second culture medium is 0.5 g/L KH2PO4，0.5g/L K2HPO4，3g/L NH4NO3，1g/L CaCl2，4g/L MgSO4And 1L distilled water, further adding 2% by volume of petroleum as a sole carbon source to the medium II, the pH of the medium II being 7.0 to 7.5.
Preferably, in S2, the porous active carrier is one or more of mesoporous silicon, mesoporous carbon, activated carbon and ceramsite.
Preferably, in S2, the bacterial liquid loading step includes: and (3) preparing a porous active carrier, namely soaking the porous active carrier in bacterial liquid, shaking for 16-20h, filtering, and loading the bacterial liquid on filler particles.
Preferably, the total amount added per liter of the petroleum wastewater is 4 x 107-8ⅹ107Thermus thermophilus of CFU.
Compared with the prior art, the invention has the beneficial effects that:
1. according to the invention, the porous active filler is used for loading the thermotrophic bacteria, so that the treatment efficiency of petroleum is improved.
2. The thermophilic thermus-inhabiting bacteria filler obtained by the invention can efficiently treat high-temperature petroleum sewage, the degradation rate of thermophilic thermus-inhabiting bacteria on petroleum is up to 72%, and compared with the prior art (50-65%), the degradation rate is higher by nearly 10%.
3. The thermophilic thermus bacterium stuffing of the invention is applied to the stoneThe components in the oil have certain degradation effect, especially on C11-C28The degradation rate of alkane is as high as more than 85 percent, and for C35The alkane degradation rate can reach 70 percent.
4. According to the invention, the treatment of high-temperature petroleum sewage is realized by only one strain (thermophagia Thermus), the degradation rate of petroleum is far higher than that of the composite strain in the prior art, and the antagonistic action among the strains in the treatment by the composite strain is avoided in the treatment of single strain.
5. The petroleum can cause harm to the environment and human bodies, so the invention can reduce the harm of the petroleum sewage to the environment and human bodies and has practical significance for maintaining the sustainable development of ecological environment.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a colony diagram of Thermus thermophilus provided in example 1 of the present invention;
FIG. 2 is a fluorescent microscopic image of Thermus thermophilus provided in example 1 of the present invention;
FIG. 3 is a graph showing the cell growth of Thermus thermophilus provided in example 1 of the present invention at 70 ℃;
FIG. 4 shows the degradation rate of petroleum in petroleum contaminated water by the method of examples 1 to 3 of the present invention;
FIG. 5 is a GC-MS spectrum of a degraded petroleum by the method of example 1 of the present invention.
Detailed Description
The following detailed description of specific embodiments of the invention is provided, but it should be understood that the scope of the invention is not limited to the specific embodiments. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention. The experimental methods described in the examples of the present invention are all conventional methods unless otherwise specified.
Example 1:
a method for treating petroleum sewage by using thermophilic bacteria specifically comprises the following steps:
(1) activation and acclimatization of Thermus thermophilus
Inoculating the Thermus thermophilus into a culture dish filled with a first culture medium for activation, culturing at 55 ℃ for 24 hours, picking out a single colony, continuously carrying out streak culture on a solid culture medium of a fresh first culture medium, and repeatedly carrying out streak inoculation culture to obtain a pure Thermus thermophilus strain;
the formula of the first culture medium is 0.5 g/L NH4NO38 g/L tryptone, 4 g/L yeast extract, 3 g/L NaCl and 1L distilled water, and the pH of the medium I was 7.0 to 7.5.
The obtained pure Thermus thermophilus colony graph is shown in figure 1, and the colony is light yellow, large and uneven in edge;
as shown in fig. 2, microscopic examination was further performed on the obtained pure thermus thermophilus, specifically: the Thermus thermophilus was stained with 6-carboxyfluorescein at room temperature for 30 minutes, washed three times with 1XPBS, and then examined under a fluorescence microscope, and it can be seen from FIG. 2 that the Thermus thermophilus emits green fluorescence under the fluorescence microscope.
Inoculating Thermus thermophilus into liquid culture medium I, culturing at 70 deg.C and 140rpm/min for 32 hr, and measuring OD at different times600The growth curve of Thermus thermophilus was plotted (FIG. 3).
The pure thermotrophic strains were then subjected to high temperature acclimatization:
in the first training stage, inoculating pure Thermus thermophilus into a second culture medium, culturing at 50 ℃ at 140rpm/min for 24 h;
in the second training stage, the thermophagia thermoacidophile which passes through the first training stage is inoculated in a second culture medium according to the inoculation amount of 10 percent, and is cultured for 24 hours at 140rpm/min and 60 ℃;
in the third training stage, the thermus thermophilus which passes through the second training stage is inoculated in a second culture medium according to the inoculation amount of 10 percent, and the second culture medium is cultured for 24 hours at the speed of 140rpm/min and the temperature of 70 ℃;
and in the fourth training stage, inoculating the thermus thermophilus subjected to the second training stage into a second culture medium according to the inoculation amount of 10%, and culturing at 140rpm/min and 80 ℃ for 24h to obtain trained thermus thermophilus.
Culturing the trained Thermus thermophilus at 80 ℃ for 24h to obtain Thermus thermophilus bacterial liquid.
The formula of the second culture medium is as follows:
0.5g/L KH2PO4，0.5g/L K2HPO4，3g/L NH4NO3，1g/L CaCl2，4g/L MgSO4and 1L distilled water, further adding 2% by volume of petroleum as a sole carbon source to the medium II, the pH of the medium II being 7.0 to 7.5.
(2) Preparation of porous active carrier
Mixing mesoporous silicon, mesoporous carbon, activated carbon and ceramsite according to the ratio of 1:1:1:1, and roasting at the high temperature of 550 ℃; leaching the filler mixture subjected to high-temperature roasting by using distilled water, and then placing the filler mixture in an oven to be dried for 24 hours at 65 ℃; then 10 is added-5Chelating in mixed solution of citric acid and ethylenediamine of/L, culturing at 30 deg.C for 24 hr under shaking, and oven drying to obtain porous active carrier.
(3) Treatment of petroleum sewage
Soaking the porous active carrier obtained in the step (2) in a Thermus thermophilus bacterial solution trained at a high temperature in the step (1), vibrating and adsorbing the solution on a shaking table for 16 hours, standing the solution for 24 hours to obtain Thermus thermophilus filler, and then putting the Thermus thermophilus filler into a reaction tank filled with petroleum sewage for treating for 48 hours, wherein the temperature of the petroleum sewage is 70 ℃;
the total amount of the above-mentioned components added per liter of petroleum wastewater is 8 x 107Thermus thermophilus of CFU.
The content of the thermoacidophilic bacteria in each liter of petroleum sewage is obtained by collecting, detecting and calculating through a conventional microorganism measuring method, and is not repeated in the invention.
Example 2:
a method for treating petroleum wastewater using thermophilic bacteria, which is substantially the same as the steps of example 1, except that: culturing the thermus thermophilus subjected to high-temperature domestication at 70 ℃ for 26h to obtain a thermus thermophilus bacterial liquid;
the porous active carrier is prepared by mixing mesoporous silicon, mesoporous carbon and active carbon in a ratio of 1:1:1, and is mixed with a bacterial solution and then is vibrated and adsorbed for 18 hours to obtain a thermophagia bacterium filler;
the high temperature acclimatization comprises:
in the first training stage, the thermophagia thermoacidophilus is inoculated in a second culture medium and cultured for 20h at the speed of 120rpm/min and the temperature of 55 ℃;
in the second training stage, inoculating the thermus thermophilus subjected to the first training stage into a second culture medium according to the inoculation amount of 8%, and culturing at the temperature of 65 ℃ for 20h at 120 rpm/min;
in the third training stage, the thermus thermophilus which passes through the second training stage is inoculated in a second culture medium according to the inoculation amount of 8 percent, and the second culture medium is cultured for 20 hours at the speed of 120rpm/min and the temperature of 75 ℃;
in the fourth training stage, the Thermus thermophilus which passes through the second training stage is inoculated into the second culture medium according to the inoculation amount of 8 percent and cultured for 20 hours at the speed of 120rpm/min and the temperature of 85 ℃.
The total amount of the additive added in per liter of petroleum sewage is 6 x 107The temperature of the petroleum sewage of the thermophagia thermoacidophiles of the CFU is 50 ℃.
Example 3:
a method for treating petroleum wastewater using thermophilic bacteria, which is substantially the same as the steps of example 1, except that: culturing the thermus thermophilus subjected to high-temperature domestication at 70 ℃ for 26h to obtain a thermus thermophilus bacterial liquid;
the porous active carrier is prepared by mixing mesoporous silicon, mesoporous carbon and active carbon in a ratio of 1:1:1, and is mixed with bacterial liquid and then is subjected to shaking adsorption for 20 hours to obtain thermophagic Thermus bacteria filler;
the high temperature acclimatization comprises:
in the first training stage, the thermophagia thermoacidophilus is inoculated in a second culture medium and cultured for 28 hours at the speed of 160rpm/min and the temperature of 53 ℃;
in the second training stage, the thermus thermophilus which passes through the first training stage is inoculated in a second culture medium according to the inoculation amount of 18 percent, and the second culture medium is cultured for 28 hours at the speed of 160rpm/min and the temperature of 63 ℃;
in the third training stage, the thermus thermophilus which passes through the second training stage is inoculated in a second culture medium according to the inoculation amount of 18 percent, and the second culture medium is cultured for 28 hours at the speed of 160rpm/min and the temperature of 73 ℃;
in the fourth training stage, the Thermus thermophilus which passes through the second training stage is inoculated into the second culture medium according to the inoculation amount of 18 percent and cultured for 28 hours at the speed of 160rpm/min and 83 ℃.
The total amount of the additive in the stone oil wastewater is 4 x 10 per liter7The temperature of the petroleum sewage is 80 ℃.
The strains used in examples 2 and 3 were the same as in example 1, and the morphological characteristics of the strains were substantially the same, and therefore, details regarding the colonies of Thermus thermophilus, microscopic examination results, and growth profiles were not repeated.
Comparative example 1:
the control was petroleum wastewater without Thermus thermophilus.
The above examples 1-3 are specific implementation steps of the method for treating petroleum sewage by using thermophilic bacteria, the petroleum contents of the examples 1-3 and the comparative example 1 are respectively detected by a gravimetric method, and the petroleum degradation rate is obtained by calculation, and the specific process is as follows:
the weight method comprises the following steps: performing oscillation extraction on petroleum sewage before and after the Thermus thermophilus treatment by using petroleum ether, wherein the dosage ratio of the petroleum ether to the petroleum sewage is 1:10, collecting an extract liquid into a separating funnel, performing oscillation standing for layering, removing a water phase, collecting an organic phase, evaporating to dryness at 65 ℃ in an oven, and weighing. And (3) calculating the total crude oil substance degradation rate according to the following calculation formula:
each letter represents a meaning: d represents the crude oil degradation rate (%); m is1Represents the residual oil amount (g) of the control group; m is2Represents the amount of residual oil after degradation (g); m is0Indicating the oil content (g) of the blank control.
The results of gravimetric method are shown in fig. 4: the degradation rate of the technical scheme in the embodiment 1 on the petroleum sewage is 76%, the degradation rate of the technical scheme in the embodiment 2 on the petroleum sewage is 66%, the degradation rate of the technical scheme in the embodiment 3 on the petroleum sewage is 74%, the average degradation rate in the embodiments 1 to 3 is up to 72%, and compared with the prior art, the degradation rate on the petroleum sewage is nearly 10% higher in the invention.
Taking example 1 as an example, the degradation condition of the Thermus thermophilus on various components of petroleum is analyzed through GC-MS detection;
the GC-MS detection adopts a Varian (30m × 0.25.25 mm × 0.25.25 μm) quartz capillary column as a chromatographic column, the detector temperature is 280 ℃, the temperature rise program is that the temperature is kept at 60 ℃ for 2min, and the temperature rise speed is 8 ℃ for ∙ min-1Heating to 280 deg.C, and keeping the temperature for 5 min; carrier gas (N)2) The flow rate is 1m L ∙ min-1The injection port temperature is 270 ℃, the injection amount is 2 mu L, and the split ratio is 20: 1 (V/V).
The detection results are shown in fig. 5: CK represents that petroleum sewage which is not treated by the thermoacidophile filler is used as a control group, and compared with the control group, the thermoacidophile (Thermus thermophilus) filler has certain degradation capability on various alkanes in petroleum and has certain degradation capability on C11-C28The degradation rate of alkane is as high as more than 85 percent, and for C35The alkane degradation rate can reach 70 percent.
It should be noted that Thermus thermophilus ATCC 33923 used in the present invention is purchased from North Nay Bio Inc.; the petroleum-polluted water is provided by Shanxi extended oil Limited liability company.
In conclusion, the invention provides a method for treating petroleum sewage by using thermophilic bacteria, the thermophilic bacteria are trained to prepare the thermophilic bacteria filler, the average degradation rate of the thermophilic bacteria filler on the petroleum sewage is up to 72 percent, and analysis shows that the technical scheme of the invention has higher degradation effect on different components in the petroleum and has higher degradation effect on C11-C28The degradation rate of alkane is as high as more than 85 percent, and for C35The alkane degradation rate can reach 70 percent.
While preferred embodiments of the present invention have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all such alterations and modifications as fall within the scope of the invention.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.
Claims (7)
1. A method for treating petroleum sewage by using thermophilic bacteria is characterized by comprising the following steps:
s1, activating thermoacidophilic bacteria, training the activated strains, and then culturing the trained strains at 70-80 ℃ for 22-26h to obtain bacterial liquid;
s2, loading the bacterial liquid obtained in the step S1 on a porous active carrier to obtain thermophage Thermus filler, and then putting the thermophage filler into petroleum wastewater for treatment, wherein the temperature of the petroleum wastewater is 50-80 ℃.
2. The method according to claim 1, wherein the culture medium for culturing said Thermus thermophilus in S1 is a first culture medium having a formula of 0.5 g/L NH4NO38 g/L tryptone, 4 g/L yeast extract, 3 g/L NaCl and 1L distilled water, and the pH of the medium I was 7.0 to 7.5.
3. The method for treating petroleum sewage according to claim 1 wherein in S1, the training comprises the following stages:
in the first training stage, the Thermus thermophilus is inoculated in a second culture medium and cultured for 20-28h at the temperature of 120-55 ℃ and at the rpm/min of 160-;
in the second training stage, the Thermus thermophilus which passes through the first training stage is inoculated in a second culture medium according to the inoculation amount of 8-18 percent, and cultured for 20-28h at the temperature of 60-65 ℃ and at the rpm of 120-;
in the third training stage, the thermus thermophilus passing through the second training stage is inoculated in a second culture medium according to the inoculation amount of 8-18 percent, and cultured for 20-28h at the temperature of 70-75 ℃ and at the rpm of 120-;
in the fourth training stage, the thermus thermophilus which passes through the second training stage is inoculated in a second culture medium according to the inoculation amount of 8-18 percent, and cultured for 20-28h at the temperature of 80-85 ℃ and at the rpm of 120-;
the second culture medium is an inorganic salt culture medium which takes petroleum as a unique carbon source.
4. The method according to claim 3, wherein the second medium is 0.5 g/L KH in the formula2PO4，0.5g/L K2HPO4，3g/L NH4NO3，1g/L CaCl2，4g/L MgSO4And 1L distilled water, further adding 2% by volume of petroleum as a sole carbon source to the medium II, the pH of the medium II being 7.0 to 7.5.
5. The method for treating petroleum sewage of claim 1 wherein in S2, the porous active carrier is one or more of mesoporous silicon, mesoporous carbon, activated carbon and ceramsite.
6. The method for treating petroleum sewage according to claim 1, wherein in S2, the bacterial liquid loading step comprises: and (3) preparing a porous active carrier, namely soaking the porous active carrier in bacterial liquid, shaking for 16-20h, filtering, and loading the bacterial liquid on the porous active carrier.
7. A method for treating petroleum sewage according to claim 1 wherein the total amount added per liter of petroleum sewage is 4 x 107-8ⅹ107Thermus thermophilus of CFU.
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Inventor after: Li Haijuan Inventor after: Zhao Jing Inventor after: Cao Jia Inventor after: Duan Xiaobin Inventor after: Chen Yueli Inventor before: Li Haijuan |