Automatic screening system for soil organic pollutant degrading bacteria

Abstract

The invention discloses an automatic screening system for soil organic pollutant degrading bacteria, which comprises a soil pretreatment module used for pretreating soil containing the degrading bacteria to obtain an aqueous solution containing the degrading bacteria and performing aeration treatment on the aqueous solution to fully consume organic matters and promote the growth of the degrading bacteria; the microorganism enrichment module is used for performing steam sterilization on an enrichment culture medium and an enrichment culture medium containing agar, and enriching degrading bacteria through the enrichment culture medium; the bacterial colony separation module is used for separating the enriched degrading bacteria on a sterilized agar-containing enrichment culture medium and positioning a target strain; the bacterial colony enrichment module is input into an enrichment culture medium containing agar from the microorganism enrichment module and is used for putting the target strains into the enrichment culture medium for enrichment culture so as to obtain a plurality of target strains. The system provided by the invention can accurately screen out dominant strains and simplify the process, so that the manual operation time and cost can be greatly reduced, and the safety of operators can be guaranteed.

Description

Technical Field

[0001] The invention relates to the technical field of soil organic pollutant microbial treatment, and in particular to an automatic screening system for soil organic pollutant degradation bacteria. Background Art

[0002] Using dominant indigenous degrading bacteria of soil organic pollutants for bioremediation of contaminated soil is an environmentally friendly and promising treatment method. They secrete various hydrolases to transform complex organic pollutants into simple inorganic substances through biochemical transformation from macromolecular toxic structures. How to screen out dominant indigenous degrading bacteria of soil organic pollutants is the most critical link in the whole process. Since soil is the gathering place of water pollution and air pollution, soil organic pollutants are classified in a complex way, including polycyclic aromatic hydrocarbons, organic pesticides, phenols, etc. The action mechanisms of different types of pollutants are very different, and specific specialized degrading bacteria are needed to deal with them. In addition, indigenous bacteria are highly adaptable to the environment, avoiding the ecological risks that may be brought by foreign microorganisms and ensuring ecological safety.

[0003] The principle of screening dominant indigenous degrading bacteria is to take an appropriate amount of soil from the contaminated site and first enrich the bacteria that degrade the pollutant in a culture medium containing the target pollutant, and then further separate, purify and culture to obtain a single dominant strain. The dominant bacteria show high activity under the target organic pollutants, accelerate the decomposition and transformation of organic pollutants, and thus effectively purify the soil.

[0004] Traditional manual screening methods are affected by multiple factors. Different pollutants require matching corresponding parameters, culture media and other environmental conditions to screen out ideal strains. However, a large number of mechanical and repetitive operations are time-consuming and labor-intensive, and the cost of microbial remediation is greatly increased. Summary of the invention

[0005] The present invention aims to solve at least one of the technical problems in the related art to a certain extent. To this end, the first purpose of the present invention is to propose an automatic screening system for soil organic pollutant-degrading bacteria, which can accurately screen out dominant strains and simplify the process, thereby greatly reducing the time and cost of manual operation, and can effectively reduce the probability of human contact with toxic pollutants, thereby ensuring the safety of operators, and solving the technical problems of traditional manual screening methods, such as cumbersome steps, a large amount of repetitive work, large influence of external factors, and limited effect.

[0006] To achieve the above-mentioned purpose, the first embodiment of the present invention proposes an automatic screening system for soil organic pollutant degrading bacteria, comprising: a soil pretreatment module, the soil pretreatment module is used to pretreat soil containing degrading bacteria to obtain an aqueous solution containing degrading bacteria, and aerate the aqueous solution to fully consume organic matter and promote the growth of degrading bacteria; a microbial enrichment module, the microbial enrichment module is connected to the soil pretreatment module, the microbial enrichment module is equipped with an enrichment medium and an enrichment medium containing agar, and the microbial enrichment module is used to enrich the enrichment medium and the enrichment medium containing agar. The collection culture medium is steam sterilized, and the degradation bacteria are enriched by the enrichment culture medium; a colony separation module, the colony separation module is connected to the microbial enrichment module, and the colony separation module is used to separate the enriched degradation bacteria on the sterilized agar-containing enrichment culture medium and locate the target strain; a colony enrichment module, the colony enrichment module is respectively connected to the microbial enrichment module and the colony separation module, and the colony enrichment module inputs the enrichment culture medium from the microbial enrichment module, and is used to place the target strain in the enrichment culture medium for enrichment culture, so as to obtain a plurality of the target strains.

[0007] According to the automatic screening system for soil organic pollutant degradation bacteria of the embodiment of the present invention, when screening the organic pollutant degradation bacteria in the soil, the soil containing the degradation bacteria is first placed in the soil pretreatment module, and sterile water is added to perform pretreatment to obtain an aqueous solution containing the degradation bacteria, and then the aqueous solution is aerated to fully consume organic matter and promote the growth of degradation bacteria. Then, the microbial enrichment module first steam sterilizes the enrichment medium and the enrichment medium containing agar, and passes the aqueous solution containing the degradation bacteria into the enrichment medium to enrich the degradation bacteria, and at the same time, the enrichment medium containing agar is transported to the colony separation module, and then the colony separation module evenly atomizes the quantitative organic pollutants on the surface of the cooled agar enrichment medium in a highly dispersed state. After the enrichment of the degradation bacteria is completed, the bacterial solution is mixed with sterile water for dilution, and the diluted bacterial solution is coated on the enrichment medium containing agar in the colony separation module to allow the degradation bacteria to grow; after the degradation bacteria grow, the colony separation module selects the degradation bacteria strain with good growth, that is, the target strain, and puts the target strain into the enrichment medium of the colony enrichment module for further enrichment and cultivation, so that multiple target strains can be obtained. Therefore, the system can accurately screen out the dominant strains and simplify the process, thereby greatly reducing the time and cost of manual operation, and can effectively reduce the probability of human contact with toxic pollutants, thereby ensuring the safety of the operator.

[0008] In addition, the automatic screening system for soil organic pollutant-degrading bacteria according to the above embodiment of the present invention may also have the following additional technical features:

[0009] According to one embodiment of the present invention, the soil pretreatment module includes: a pretreatment box; an inlet, which is arranged on one side of the top of the pretreatment box; a drive motor, which is arranged at the center of the top of the pretreatment box; a first agitator, which is arranged in the pretreatment box and connected to the drive motor; a screen, which is arranged at the bottom of the pretreatment box and is spaced a preset distance from the first agitator; an aeration box, which is connected to the pretreatment box by a first bolt; a blower, which is connected to an air duct, which is arranged on one side of the aeration box and extends into it; a liquid level meter, which is used to monitor the liquid level in the aeration box.

[0010] According to one embodiment of the present invention, the microorganism enrichment module includes: a first culture tank, the first culture tank is connected to the aeration box through a pipeline, the pipeline passes through a first rotary sealing plug and leads to the interior of the first culture tank, a first plunger pump and a first check valve are arranged between the first culture tank and the aeration box; a first culture medium pouring port is arranged on the top of the first culture tank, and the first culture medium pouring port is suitable for being sealed by a first sealing cover; the first culture tank is used to fill the enrichment culture medium; a second agitator, the second agitator is arranged in the first culture tank; a second culture tank, the top of the second culture tank is provided with a second culture medium pouring port, the second culture medium pouring port is suitable for being sealed by a second sealing cover The invention relates to a novel culture tank comprising a first culture tank and a second culture tank, wherein the first culture tank is provided with a spiral pipeline, and the first culture tank is provided with a second culture tank. The first culture tank is provided with a spiral pipeline, and the first culture tank is provided with a second culture tank. The first culture tank is provided with a spiral pipeline, and the first culture tank is provided with a second culture tank. The first culture tank is provided with a first booster pump and a first ball valve. The second culture tank is provided with a second booster pump and a second ball valve.

[0011] According to one embodiment of the present invention, the first culture tank is equipped with a first base, the first base is connected to the first culture tank via a first rotating shaft, and the first base includes a first cross bar; the second culture tank is equipped with a second base, the second base is connected to the second culture tank via a second rotating shaft, and the second base includes a second cross bar.

[0012] According to one embodiment of the present invention, a first pressure gauge, a first temperature sensor and a first exhaust valve are disposed outside the first culture tank; a second pressure gauge, a second temperature sensor and a second exhaust valve are disposed outside the second culture tank.

[0013] According to one embodiment of the present invention, the colony separation module includes: an incubator, the incubator is connected to the steam generator, and a third booster pump and a third ball valve are arranged between the incubator and the steam generator; a first water distributor, the first water distributor is connected to the second culture tank through a pipeline, the pipeline passes through a second rotary sealing plug and leads to the interior of the second culture tank, and a second plunger pump and a second check valve are arranged between the first water distributor and the second culture tank; a plurality of flat plates, the plurality of flat plates are circular and evenly distributed on a turntable, a third rotating shaft is arranged at the center of the turntable, the third rotating shaft is connected to a motor, a fixing frame is arranged on the turntable, and the fixing frame is used to fix the flat plate; the flat plate is suitable for being sealed by a flat plate cover, and the flat plate cover is controlled by an electric suction cup; a dilution coating group Part, one end of the dilution and coating component is connected to the first culture tank through a pipeline, the pipeline passes through the third rotary sealing plug and leads to the interior of the first culture tank, a precision pump and a third check valve are arranged between the dilution and coating component and the first culture tank, the precision pump and the third booster pump are connected through a pipeline, and a fourth ball valve is arranged between the precision pump and the third booster pump; the other end of the dilution and coating component is connected to the waste liquid barrel and the sterile water barrel through pipelines respectively, a precision circulation pump and an eighth check valve are arranged between the dilution and coating component and the waste liquid barrel, and a fourth check valve is arranged between the waste liquid barrel and the sterile water barrel; an atomizer, a filter is arranged at the bottom end of the atomizer, and an atomizing nozzle is arranged at the front end of the atomizer, which is used to make the sprayed quantitative organic pollutant liquid highly dispersed.

[0014] According to one embodiment of the present invention, a colony counter and a robotic arm are provided in the incubator, and the colony counter has the function of magnifying and imaging the colonies on the surface of the plate to locate the target strain; the robotic arm includes a fourth rotating shaft, a rotating wheel, a first telescopic rod and a pliers to select the target strain; the incubator is connected to a heating fan, and the heating fan is used to adjust the temperature of the incubator, and a fifth check valve is provided between the incubator and the heating fan; the incubator is connected to a vacuum pump, and the vacuum pump is used to control the presence or absence of oxygen in the incubator, and a sixth check valve is provided between the incubator and the vacuum pump; the incubator is also provided with a third pressure gauge, a third temperature sensor and a third exhaust valve.

[0015] According to one embodiment of the present invention, the dilution and coating assembly includes: a second telescopic rod; a mixing dish, the mixing dish is connected to the second telescopic rod by a second bolt, the mixing dish is used to mix the liquid pumped into the first culture tank and the sterile water bucket; a rotating sealing pipe, the rotating sealing pipe is connected to an opening on one side of the mixing dish; a dilution coating device, the dilution coating device is connected to the rotating sealing pipe, and the bottom of the dilution coating device is provided with uniform and dense small holes; a small round rod, the small round rod is arranged below the rotating sealing pipe and fixedly connected to the rotating sealing pipe; a hook, the hook is suitable for being connected to the small round rod; a traction rope, the traction rope is connected to the hook, and the traction rope is connected to a fixed pulley.

[0016] According to one embodiment of the present invention, the colony enrichment module includes: a telescopic pipe fitting, which is connected to the second check valve through a pipeline, and a seventh check valve is provided between the telescopic pipe fitting and the second check valve; a second water distributor, which is connected to the telescopic pipe fitting, and has multiple liquid outlets; a fermentation tank, which is multiple and arranged below the liquid outlet; a rotating rack, which is distributed in multiple layers and the fermentation tank is fixed to the table top of the rotating rack by a fixing clamp.

[0017] According to one embodiment of the present invention, each layer of the rotating frame is provided with a plurality of independent table surfaces; the rotating frame is controlled to rotate by a fifth rotating shaft; an electromagnet is provided at the upper end of each layer of the table surface of the rotating frame, and the electromagnet is wrapped in a sealing sleeve; a magnetic material is provided around the top of the lid of the fermentation tank; the size of the magnetic material circle of the lid of the fermentation tank is consistent with the size of the electromagnet, and the center positions of the circles coincide.

[0018] The automatic screening system for soil organic pollutant-degrading bacteria according to the embodiment of the present invention has the following beneficial effects:

[0019] 1. The present invention simplifies the process by setting up automatic screening of soil organic pollutant-degrading bacteria, greatly reducing manual operation time and cost, and the process is simple and easy to operate. Traditional manual methods may bring about artificial contamination bacteria and manual operation errors. The automatic screening system is more conducive to accurately screening out dominant strains. The automatic screening system can produce a variety of dominant strains in batches at one time, greatly diversifying the strain output types and improving production efficiency.

[0020] 2. The automatic screening system for soil organic pollutant degradation bacteria set by the present invention is applicable to diversified soil organic pollutants and has universal applicability. In the face of complex soil organic pollutants, the system parameters can be automatically adjusted to achieve strain screening, including dilution gradient, dilution coating usage, incubator temperature, and the presence or absence of oxygen.

[0021] 3. The present invention effectively reduces the probability of human contact with toxic pollutants. In addition, some degrading bacteria may be harmful to the human body, and the automatic screening system ensures the safety and health of the operator.

[0022] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 Schematic diagram of a block diagram of an automatic screening system for soil organic pollutant-degrading bacteria according to an embodiment of the present invention;

[0024] Figure 2 Schematic diagram of the overall structure of an automatic screening system for soil organic pollutant-degrading bacteria according to an embodiment of the present invention;

[0025] Figure 3 is a schematic diagram of the cross-sectional structure of an incubator according to an embodiment of the present invention;

[0026] Figure 4 A schematic diagram of the structure of a dilution coating assembly according to an embodiment of the present invention;

[0027] Figure 5 is a schematic structural diagram of a flat plate, a turntable and other structures according to an embodiment of the present invention;

[0028] Figure 6 A schematic diagram of the structure of a flat plate cover and an electric suction cup according to an embodiment of the present invention;

[0029] Figure 7 Schematic diagram of the structure of a fermentation tank, a rotating rack and the like according to an embodiment of the present invention.

[0030] Reference numerals:

[0031] 100, soil pretreatment module; 110, pretreatment box; 101, sample inlet; 102, drive motor; 103, first agitator; 104, screen; 105, first bolt; 106, liquid level meter; 107, blower; 108, first plunger pump; 109, first check valve; 111, aeration box;

[0032] 200, microorganism enrichment module; 201, first culture tank; 202, second culture tank; 203, heater; 204, steam generator; 205, second booster pump; 206, third booster pump; 207, first base; 208, first rotating shaft; 209, first booster pump; 210, first ball valve; 211, first exhaust valve; 212, first sealing cover; 213, first rotary sealing plug; 214, second agitator; 215, third rotary sealing plug ; 216, first pressure gauge; 217, first temperature sensor; 218, second ball valve; 219, second exhaust valve; 220, third agitator; 222, second rotating shaft; 223, second base; 224, second cross bar; 225, second temperature sensor; 226, second pressure gauge; 227, second plunger pump; 228, second check valve; 229, third ball valve; 230, second sealing cover; 232, second rotary sealing plug; 233, first cross bar;

[0033] 300, colony separation module; 301, incubator; 302, seventh check valve; 303, precision pump; 304, third check valve; 305, fourth ball valve; 306, dilution coating assembly; 307, precision circulation pump; 308, eighth check valve; 309, waste liquid bucket; 310, sterile water bucket; 311, first water distributor; 312, flat plate; 313, third pressure gauge; 314, third temperature sensor; 315, third exhaust valve; 316, filter; 317, nozzle; 318, colony counter; 319, fourth shaft; 320, robotic arm; 321, rotating wheel; 3 22. first telescopic rod; 323. pliers; 324. sixth check valve; 325. vacuum pump; 326. heating fan; 327. fifth check valve; 328. picker box; 329. second telescopic rod; 330. second bolt; 331. mixing dish; 332. diluting applicator; 333. small round rod; 334. rotating sealing pipe fitting; 335. fixed pulley; 336. third telescopic rod; 337. electric suction cup; 338. flat cover; 339. fixing frame; 340. turntable; 341. third rotating shaft; 342. atomizer; 343. fourth check valve; 344. hook;

[0034] 400. Colony enrichment module; 401. Fermentation tank; 402. Telescopic pipe; 403. Second water distributor; 404. Fifth rotating shaft; 405. Fixing clamp; 406. Cover; 407. Electromagnet; 408. Rotating frame. DETAILED DESCRIPTION

[0035] Embodiments of the present invention are described in detail below, examples of which are shown in the accompanying drawings, wherein the same or similar reference numerals throughout represent the same or similar elements or elements having the same or similar functions. The embodiments described below with reference to the accompanying drawings are exemplary and are intended to be used to explain the present invention, and should not be construed as limiting the present invention.

[0036] The automatic screening system for soil organic pollutant-degrading bacteria proposed in an embodiment of the present invention will be described below with reference to the accompanying drawings.

[0037] like Figure 1 As shown, the automatic screening system for soil organic pollutant-degrading bacteria according to an embodiment of the present invention comprises: a soil pretreatment module 100 , a microorganism enrichment module 200 , a colony separation module 300 and a colony enrichment module 400 .

[0038] Wherein, the soil pretreatment module 100 is used to pretreat the soil containing degradation bacteria, obtain an aqueous solution containing degradation bacteria, and aerate the aqueous solution to fully consume organic matter and promote the growth of degradation bacteria. The microbial enrichment module 200 is connected to the soil pretreatment module 100, and the microbial enrichment module 200 is equipped with an enrichment medium and an enrichment medium containing agar. The microbial enrichment module 200 is used to steam sterilize the enrichment medium and the enrichment medium containing agar, and enrich the degradation bacteria through the enrichment medium. The colony separation module 300 is connected to the microbial enrichment module 200, and the colony separation module 300 is used for enrichment medium. The colony enrichment module 400 is respectively connected to the microbial enrichment module 200 and the colony separation module 300, and the colony enrichment module 400 is input with the enrichment medium by the microbial enrichment module 200, and is used to put the target strain into the enrichment medium for enrichment culture to obtain multiple target strains.

[0039] Specifically, when screening organic pollutant degrading bacteria in soil, firstly, the soil containing degrading bacteria is placed in the soil pretreatment module 100, and sterile water is added for pretreatment to obtain an aqueous solution containing degrading bacteria, and then the aqueous solution is aerated to fully consume organic matter and promote the growth of degrading bacteria. Then, the microbial enrichment module 200 first steam sterilizes the enrichment medium and the enrichment medium containing agar, respectively, and passes the aqueous solution containing degrading bacteria into the enrichment medium to enrich the degrading bacteria, and at the same time, the enrichment medium containing agar is transported to the colony separation module 300, and then the colony separation module 300 uniformly atomizes the quantitative organic pollutants on the surface of the cooled agar enrichment medium in a highly dispersed state. After the enrichment of the degrading bacteria is completed, the bacterial solution is mixed with sterile water for dilution, and the diluted bacterial solution is spread on the enrichment medium containing agar in the colony separation module 300 to allow the degrading bacteria to grow; after the degrading bacteria grow, the colony separation module 300 selects the degrading bacteria strain with good growth, that is, the target strain, and puts the target strain into the enrichment medium in the colony enrichment module 400 for further enrichment culture, so that multiple target strains can be obtained.

[0040] According to one embodiment of the present invention, Figure 2 As shown, the soil pretreatment module 100 includes: a pretreatment box 110; an injection port 101, which is arranged on one side of the top of the pretreatment box 110; a drive motor 102, which is arranged at the center of the top of the pretreatment box 110; a first agitator 103, which is arranged in the pretreatment box 110 and connected to the drive motor 102; a screen 104, which is arranged at the bottom of the pretreatment box 110 and is spaced a preset distance from the first agitator 103; an aeration box 111, which is connected to the pretreatment box 110 by a first bolt 105; a blower 107, which is connected to an air duct, which is arranged on one side of the aeration box 111 and extends into it; a liquid level meter 106, which is used to monitor the liquid level height in the aeration box 111.

[0041] Specifically, in order to solve the problem of soil collected in situ containing impurities such as gravel, the soil needs to be pretreated. Specifically, the soil collected in situ enters from the sampling port 101, and an appropriate amount of sterile water is added. The driving motor 102 starts the first agitator 103, and the mud is filtered into the aeration box 111 through the screen 104. Further, the blower 107 is aerated through the air duct to fully consume organic matter and promote microbial growth, and then the soil is allowed to settle. In this process, the liquid level in the aeration box 111 is monitored in real time by the liquid level meter 106.

[0042] According to one embodiment of the present invention, Figure 2As shown, the microorganism enrichment module 200 includes: a first culture tank 201, the first culture tank 201 is connected to the aeration box 111 through a pipeline, the pipeline passes through the first rotary sealing plug 213 and enters the interior of the first culture tank 201, and a first plunger pump 108 and a first check valve 109 are arranged between the first culture tank 201 and the aeration box 111; a first culture medium pouring port is arranged on the top of the first culture tank 201, and the first culture medium pouring port is suitable for being sealed by a first sealing cover 212; the first culture tank 201 is used to fill the enrichment culture medium; a second agitator 214, and the second agitator 214 is arranged in the first culture tank 201; a second culture tank 202, and a second culture medium pouring port is arranged on the top of the second culture tank 202, and the second culture medium pouring port is suitable for being sealed by a second sealing cover 230; The second culture tank 202 is used for filling the enriched culture medium containing agar and the enriched culture medium; the third agitator 220, the third agitator 220 is arranged in the second culture tank 202; the inner wall of the first culture tank 201 and the inner wall of the second culture tank 202 are both provided with spiral pipelines, and small holes are evenly opened on the spiral pipelines; a steam generating device, the steam generating device includes a heater 203 and a steam generator 204; the steam generator 204 is respectively connected to the spiral pipeline in the first culture tank 201 and the spiral pipeline in the second culture tank 202; a first booster pump 209 and a first ball valve 210 are arranged between the steam generator 204 and the first culture tank 201; a second booster pump 205 and a second ball valve 218 are arranged between the steam generator 204 and the second culture tank 202.

[0043] According to one embodiment of the present invention, Figure 2 As shown, a first pressure gauge 216 , a first temperature sensor 217 and a first exhaust valve 211 are disposed outside the first culture tank 201 ; a second pressure gauge 226 , a second temperature sensor 225 and a second exhaust valve 219 are disposed outside the second culture tank 202 .

[0044] According to one embodiment of the present invention, Figures 2 to 6As shown, the colony separation module 300 includes: an incubator 301, the incubator 301 is connected to the steam generator 204, and a third booster pump 206 and a third ball valve 229 are arranged between the incubator 301 and the steam generator 204; a first water distributor 311, the first water distributor 311 is connected to the second culture tank 202 through a pipeline, the pipeline passes through the second rotary sealing plug 232 and enters the interior of the second culture tank 202, and the first water distributor 311 and the second culture tank 202 are connected. A second plunger pump 227 and a second check valve 228 are arranged between the plurality of flat plates 312, the plurality of flat plates 312 are circular and evenly spaced on a turntable 340, a third rotating shaft 341 is arranged at the center of the turntable 340, the third rotating shaft 341 is connected to the motor, a fixing frame 339 is arranged on the turntable 340, and the fixing frame 339 is used to fix the flat plate 312; the flat plate 312 is suitable for being sealed by a flat plate cover 338, and the flat plate cover 338 is controlled by an electric suction cup 337; a dilution coating assembly 3 06, one end of the dilution coating assembly 306 is connected to the first culture tank 201 through a pipeline, the pipeline passes through the third rotary sealing plug 215 and enters the interior of the first culture tank 201, a precision pump 303 and a third check valve 304 are arranged between the dilution coating assembly 306 and the first culture tank 201, the precision pump 303 is connected to the third booster pump 206 through a pipeline, and a fourth ball valve 305 is arranged between the precision pump 303 and the third booster pump 206; the dilution coating assembly 306 The other end is connected to the waste liquid barrel 309 and the sterile water barrel 310 through pipelines, respectively. A precision circulation pump 307 and an eighth check valve 308 are provided between the dilution coating assembly 306 and the waste liquid barrel 309, and a fourth check valve 343 is provided between the waste liquid barrel 309 and the sterile water barrel 310; an atomizer 342, a filter screen 316 is provided at the bottom end of the atomizer 342, and an atomizing nozzle is provided at the front end of the atomizer 342 to make the quantitative organic pollutant liquid sprayed out in a highly dispersed state. The incubator 301 is also provided with a third pressure gauge 313, a third temperature sensor 314 and a third exhaust valve 315.

[0045] Specifically, first, the enriched culture medium is added to the first culture tank 201, and the first sealing cover 212 is tightened, the enriched culture medium containing agar is added to the second culture tank 202, and the second sealing cover 230 is tightened, then the heater 203 is powered on, the steam generator 204 generates steam, the first booster pump 209, the second booster pump 205 and the third booster pump 206 are turned on, the first ball valve 210, the second ball valve 218, the third ball valve 229, and the fourth ball valve 305 are opened, and the pressure and temperature in the first culture tank 201, the second culture tank 202, and the incubator 301 are monitored by the first pressure gauge 216, the second pressure gauge 226, the third pressure gauge 313 and the first temperature sensor 217, the second temperature sensor 225, and the third temperature sensor 314, respectively, so that the first culture tank 201, the second culture tank 202, the incubator 301, the first water distributor 311, the dilution coating assembly 306 and the related connecting pipes are fully sterilized.

[0046] After the sterilization is completed, the first booster pump 209, the second booster pump 205 and the third booster pump 206 are closed, the first ball valve 210, the second ball valve 218, the third ball valve 229, the fourth ball valve 305 are opened, and the first exhaust valve 211, the second exhaust valve 219, and the third exhaust valve 315 are opened to release the pressure. The second plunger pump 227 is turned on, and the second check valve 228 is opened. The agar enrichment medium in the second culture tank 202 passes through the first water distributor 311 while hot, and the liquid enters the flat plate 312 in the form of horizontal projection motion, and solidifies after cooling. The atomizer 342 uniformly atomizes a certain amount of organic pollutants on the surface of the cooled agar enrichment medium in a highly dispersed state, wherein the filter 316 can intercept the interference of foreign bacteria in the air, and the atomizing nozzle 317 can spray different types of organic pollutants in a highly dispersed state. Then, the enrichment medium is added to the second culture tank 202, and the sterilization operation is completed again, and cooled for standby use.

[0047] Furthermore, when the liquid level in the aeration tank 111 reaches the specified height of the liquid level meter 106, the first plunger pump 108 is turned on, the first check valve 109 is opened, and the supernatant in the aeration tank 111 enters the first culture tank 201 for microbial (degradation bacteria) enrichment culture. When the set culture time is reached, the precision pump 303 is turned on, the third check valve 304 is opened, and the bacterial solution is quantitatively sucked into the dilution coating component 306. At the same time, the precision circulation pump 307 is turned on, the eighth check valve 308 is opened, and the sterile water is quantitatively sucked and mixed with the bacterial solution by convection, and the turntable 340 is rotated, and the dilution coating component 306 performs a streaking separation operation on the flat plate 312.

[0048] Furthermore, different bacterial solution concentration gradient parameters can be set, the precision circulation pump 307, the eighth check valve 308 is closed, the fourth check valve 343 is opened, and after the excess bacterial solution is quantitatively extracted, the eighth check valve 308 is opened, the fourth check valve 343 is closed, and sterile water is quantitatively sucked into the dilution coating component 306, and the streaking separation operation is repeated on another plate 312. In this way, multiple (such as 6) gradient bacterial solution concentrations can be operated to achieve the purpose of thorough separation and purification.

[0049] Specifically, when the bottom end of the dilution coating assembly 306 contacts the agar-enriched culture medium on the plate 312, the purpose of streaking and separating the strains is achieved by rotating the turntable 340. The turntable 340 is provided with a fixing frame 339 to fix the plate 312, and the center of the turntable 340 has a third rotating shaft 341. After the streaking and separation operations are performed one by one on the agar-enriched culture medium on the plate 312, the third rotating shaft 341 returns the turntable 340 to the initial position, and the electric suction cup 337 activates the third telescopic rod 336, and the electric suction cup 337 firmly sucks the plate cover 338, covers the plate cover 338 on the plate 312, and then the electric suction cup 337 is disengaged and returns to the initial position.

[0050] It should be understood that the steam pipes of the first culture tank 201 and the second culture tank 202 are spirally distributed around the tank body, and small holes are evenly distributed on the pipes. The second stirrer 214 and the third stirrer 220 serve the purpose of stirring and evenly heating. The inlet and outlet pipe connection ports of the first culture tank 201 and the second culture tank 202 are sealed with special rotary sealing plugs, namely the first rotary sealing plug 213, the second rotary sealing plug 232, and the third rotary sealing plug 215, which can better play a sealing role.

[0051] According to one embodiment of the present invention, Figure 2 As shown, the first culture tank 201 is equipped with a first base 207, which is connected to the first culture tank 201 through a first rotating shaft 208, and the first base 207 includes a first crossbar 233; the second culture tank 202 is equipped with a second base 223, which is connected to the second culture tank 202 through a second rotating shaft 222, and the second base 223 includes a second crossbar 224. Therefore, when cleaning the first culture tank 201 or the second culture tank 202, the tank body can be turned over by the first rotating shaft 208 or the second rotating shaft 222 for easy cleaning.

[0052] According to one embodiment of the present invention, Figure 3As shown, a colony counter 318 and a mechanical arm 320 are provided in the incubator 301. The colony counter 318 has the function of amplifying the colonies on the surface of the imaging plate 312 for locating the target strain; the mechanical arm 320 includes a fourth rotating shaft 319, a rotating wheel 321, a first telescopic rod 322 and a forceps 323 for selecting the target strain; the incubator 301 is connected to a heating fan 326, the heating fan 326 is used to adjust the temperature of the incubator 301, and a fifth check valve 327 is provided between the incubator 301 and the heating fan 326; the incubator 301 is connected to a vacuum pump 325, the vacuum pump 325 is used to control the presence or absence of oxygen in the incubator 301, and a sixth check valve 324 is provided between the incubator 301 and the vacuum pump 325.

[0053] Specifically, the colony counter 318 has the function of magnifying the colonies on the surface of the imaging plate 312, and can select the number and position of colonies with good growth. Each plate 312 uses the center of its surface as the pole to establish a polar coordinate system to indicate the preferred colony position. The mechanical arm 320 includes a rotating shaft, a rotating wheel 321, a first telescopic rod 322 and a pliers 323, wherein the rotating shaft and the rotating wheel 321 respectively control the mechanical arm 320 to move up and down and rotate freely, so as to facilitate the precise positioning of the pliers 323 and clamp the picker of the colony picker box 328.

[0054] Furthermore, the heating fan 326 can adjust the temperature of the incubator 301 so that the inside of the incubator 301 reaches a constant temperature; the vacuum pump 325 can draw a vacuum, control the presence or absence of oxygen inside the incubator 301, and screen aerobic bacteria or anaerobic bacteria.

[0055] According to one embodiment of the present invention, Figure 4 As shown, the dilution coating assembly 306 includes: a second telescopic rod 329; a mixing dish 331, which is connected to the second telescopic rod 329 through a second bolt 330, and the mixing dish 331 is used to mix the liquid pumped into the first culture tank 201 and the sterile water bucket 310; a rotating sealing pipe 334, which is connected to a hole on one side of the mixing dish 331; a dilution coating device 332, which is connected to the rotating sealing pipe 334, and the bottom of the dilution coating device 332 is distributed with uniform and dense small holes; a small round rod 333, which is arranged below the rotating sealing pipe 334 and fixedly connected to the rotating sealing pipe 334; a hook 344, which is suitable for being connected to the small round rod 333; a traction rope, which is connected to the hook 344, and the traction rope is connected to the fixed pulley 335.

[0056] Specifically, the traction rope pulls the small round rod 333 through the fixed pulley 335, and the rotatable sealing pipe 334 rotates 180° clockwise, so that the bottom end of the dilution applicator 332 faces upward. After the two liquids pumped from the first culture tank 201 and the sterile water bucket 310 are evenly mixed in the mixing dish 331, the traction rope pulls the small round rod 333 through the fixed pulley 335, and the rotatable sealing pipe 334 rotates 180° counterclockwise, so that the bottom end of the dilution applicator 332 faces downward. The liquid of a certain height in the mixing dish 331 flows into the dilution applicator 332, and the second telescopic rod 329 controls the dilution applicator 332 to contact the agar-enriched culture medium of the flat plate 312. The mixing dish 331 can be customized according to the amount of bacterial liquid quantitatively flowing into the dilution applicator 332. The second bolt 330 facilitates disassembly.

[0057] According to one embodiment of the present invention, Figure 2 and Figure 7 As shown, the colony enrichment module 400 includes: a telescopic pipe fitting 402, which is connected to the second check valve 228 through a pipeline, and a seventh check valve 302 is provided between the telescopic pipe fitting 402 and the second check valve 228; a second water distributor 403, which is connected to the telescopic pipe fitting 402, and has multiple liquid outlets; a fermentation tank 401, which is multiple and arranged below the liquid outlet; a rotating rack 408, which is distributed in multiple layers and is fixed to the table of the rotating rack 408 by a fixing clamp 405.

[0058] Specifically, after the target strain is separated, the enrichment medium is added to the second culture tank 202, and the second sealing cover 230 is tightened, the heater 203 is powered on again, the steam generator 204 generates steam, the second booster pump 205 is turned on, and the second ball valve 218 is opened to sterilize the second culture tank 202. After the sterilization is completed, the second plunger pump 227 is turned on, the second check valve 228 and the seventh check valve 302 are opened, so that the sterilized enrichment medium passes through the second water distributor 403 and enters the multiple fermentation tanks 401. Then the robotic arm 320 puts the target strain into the fermentation tank 401 for further enrichment culture, and finally multiple dominant strains can be obtained.

[0059] Furthermore, after the second water distributor 403 completes one liquid distribution, the telescopic pipe 4024 can control the second water distributor 403 to become shorter, and after the rotating frame 408 rotates the empty fermentation tank 401 on the other side, the telescopic pipe 402 can control the second water distributor 403 to become longer to continue pumping in the enriched culture medium.

[0060] According to one embodiment of the present invention, Figure 7As shown, each layer of the rotating frame 408 is provided with multiple independent countertops; the rotating frame 408 is controlled to rotate by the fifth rotating shaft 404; an electromagnet 407 is provided at the upper end of each countertop of the rotating frame 408, and the electromagnet 407 is wrapped in a sealing sleeve, and a magnetic material is provided around the top of the lid 406 of the fermentation tank 401, and the size of the magnetic material circle of the lid 406 of the fermentation tank 401 is consistent with the size of the electromagnet 407, and the center position of the circle coincides. In a specific example, the rotating frame 408 is distributed in six layers, and each layer of the rotating frame 408 is provided with four independent countertops.

[0061] Specifically, after the target strain is placed in the fermentation tank 401 , the electromagnet 407 controls the lid 406 of the fermentation tank 401 to be lowered and covered on the fermentation tank 401 to seal the fermentation tank 401 .

[0062] Combine the following Figures 2 to 7 , describes in detail the specific working process of the automatic screening system for soil organic pollutant-degrading bacteria in an embodiment of the present invention.

[0063] The automatic screening system for soil organic pollutant-degrading bacteria according to the embodiment of the present invention may include the following steps:

[0064] S1. Collect the contaminated soil and pour it into the pretreatment box 110, and then pour in sterile water. Under the action of the first agitator 103, the mud penetrates into the aeration box 111 through the screen 104, and is aerated to fully consume organic matter and promote the growth of microorganisms, and then is allowed to settle.

[0065] S2. Add the enriched medium and the enriched medium containing agar to the first culture tank 201 and the second culture tank 202 respectively, turn on the steam generator 204, and fully steam sterilize the first culture tank 201, the second culture tank 202, the incubator 301 and the pipeline. The first culture tank 201 is cooled for standby use; the enriched medium containing agar in the second culture tank 202 is pumped into the flat plate 312 while hot to cool and solidify. Then, the enriched medium is added to the second culture tank 202, and the sterilization operation is completed again, and the mixture is cooled for standby use.

[0066] S3. When the liquid level in the aeration tank 111 reaches the detection height of the liquid level meter 106, the first plunger pump 108 is turned on, and the supernatant in the aeration tank 111 is pumped into the first culture tank 201 for microbial enrichment culture.

[0067] S4, quantitatively extract the bacterial solution from the first culture tank 201 and mix it with sterile water for dilution, and six dilution gradients can be selected. After quantitative control, the diluted bacterial solution is evenly distributed at the bottom of the dilution applicator 332, and the plate 312 is rotated to achieve the purpose of dispersing the bacterial solution. The electric suction cup 337 controls the plate cover 338 to be put down and taken up. The vacuum pump 325 can control the oxygen in the incubator 301, and selectively screen out aerobic bacteria or anaerobic bacteria.

[0068] S5. After the colonies grow, the surface of the imaging plate 312 is magnified, the number of colonies with good growth is selected, and the number of fermentation tanks 401 is determined at the same time, and the culture medium of the second culture tank 202 is pumped into the corresponding fermentation tank 401. According to the coordinate positioning, the robot arm 320 selects the colonies and puts them into the fermentation tank 401 for enrichment culture. The rotating frame 408 operates multiple fermentation tanks 401 in batches, and the electromagnet 407 controls the lid of the fermentation tank 401 to be lowered, and finally multiple dominant strains are obtained.

[0069] In summary, according to the automatic screening system for soil organic pollutant degradation bacteria of an embodiment of the present invention, when screening organic pollutant degradation bacteria in soil, the soil containing degradation bacteria is first placed in a soil pretreatment module, and sterile water is added for pretreatment to obtain an aqueous solution containing degradation bacteria, and then the aqueous solution is aerated to fully consume organic matter and promote the growth of degradation bacteria. Then, the microbial enrichment module first steam sterilizes the enrichment medium and the enrichment medium containing agar, respectively, and passes the aqueous solution containing degradation bacteria into the enrichment medium to enrich the degradation bacteria, and at the same time, the enrichment medium containing agar is transported to the colony separation module, and then the colony separation module evenly atomizes the quantitative organic pollutants on the surface of the cooled agar enrichment medium in a highly dispersed state. After the enrichment of the degradation bacteria is completed, the bacterial solution is mixed with sterile water for dilution, and the diluted bacterial solution is coated on the enrichment medium containing agar in the colony separation module to allow the degradation bacteria to grow; after the degradation bacteria grow, the colony separation module selects the degradation bacteria strain with good growth, that is, the target strain, and puts the target strain into the enrichment medium of the colony enrichment module for further enrichment and cultivation, so that multiple target strains can be obtained. Therefore, the system can accurately screen out the dominant strains and simplify the process, thereby greatly reducing the time and cost of manual operation, and can effectively reduce the probability of human contact with toxic pollutants, thereby ensuring the safety of the operator.

[0070] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "examples", "specific examples", or "some examples" means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner.

[0071] In addition, the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include at least one of the features. In the description of the present invention, the meaning of "plurality" is at least two, such as two, three, etc., unless otherwise clearly and specifically defined.

[0072] In the present invention, unless otherwise clearly specified and limited, the terms "installed", "connected", "connected", "fixed" and the like should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements, unless otherwise clearly defined. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0073] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. An automatic screening system for soil organic pollutant-degrading bacteria, characterized in that: include: A soil pretreatment module, which is used to pretreat the soil containing degrading bacteria to obtain an aqueous solution containing degrading bacteria, and to aerate the aqueous solution to fully consume organic matter and promote the growth of degrading bacteria; A microbial enrichment module, wherein the microbial enrichment module is connected to the soil pretreatment module, wherein an enrichment medium and an enrichment medium containing agar are contained in the microbial enrichment module, wherein the microbial enrichment module is used to steam sterilize the enrichment medium and the enrichment medium containing agar, and to enrich degradation bacteria through the enrichment medium; A colony separation module, which is connected to the microorganism enrichment module and is used to separate the enriched degrading bacteria on a sterilized agar-containing enrichment medium and locate the target strain; A colony enrichment module, wherein the colony enrichment module is connected to the microorganism enrichment module and the colony separation module respectively, and the colony enrichment module is input with an enrichment medium from the microorganism enrichment module, and is used to place the target strain into the enrichment medium for enrichment culture to obtain a plurality of the target strains.

2. The automatic screening system for soil organic pollutant-degrading bacteria according to claim 1, characterized in that: The soil pretreatment module comprises: Pre-treatment box; A sample inlet, which is arranged on one side of the top of the pretreatment box; A driving motor, wherein the driving motor is arranged at the center of the top of the pretreatment box; A first agitator, which is disposed in the pretreatment box and connected to the drive motor; A screen, the screen is arranged at the bottom of the pretreatment box and is spaced a preset distance from the first agitator; an aeration box, wherein the aeration box is connected to the pretreatment box via a first bolt; A blower, the blower is connected to an air duct, the air duct is arranged on one side of the aeration box and extends into the aeration box; A liquid level meter is used to monitor the liquid level in the aeration tank.

3. The automatic screening system for soil organic pollutant-degrading bacteria according to claim 2 is characterized in that: The microbial enrichment module comprises: a first culture tank, the first culture tank is connected to the aeration box through a pipeline, the pipeline passes through a first rotary sealing plug and leads into the interior of the first culture tank, and a first plunger pump and a first check valve are arranged between the first culture tank and the aeration box; The top of the first culture tank is provided with a first medium pouring port, and the first medium pouring port is suitable for being sealed by a first sealing cover; the first culture tank is used to be filled with enriched medium; a second agitator, the second agitator being disposed in the first culture tank; A second culture tank, wherein the top of the second culture tank is provided with a second culture medium pouring port, and the second culture medium pouring port is suitable for being sealed by a second sealing cover; the second culture tank is used for filling the enriched culture medium containing agar and the enriched culture medium; a third agitator, the third agitator being disposed in the second culture tank; The inner wall of the first culture tank and the inner wall of the second culture tank are both provided with a spiral pipeline, and small holes are evenly opened on the spiral pipeline; A steam generating device, the steam generating device comprising a heater and a steam generator; The steam generator is connected to the spiral pipeline in the first culture tank and the spiral pipeline in the second culture tank respectively; A first booster pump and a first ball valve are provided between the steam generator and the first culture tank; A second booster pump and a second ball valve are provided between the steam generator and the second culture tank.

4. The automatic screening system for soil organic pollutant-degrading bacteria according to claim 3 is characterized in that: The first culture tank is provided with a first base, the first base is connected to the first culture tank via a first rotating shaft, and the first base includes a first crossbar; The second culture tank is installed with a second base, the second base is connected to the second culture tank via a second rotating shaft, and the second base includes a second cross bar.

5. The automatic screening system for soil organic pollutant-degrading bacteria according to claim 3 is characterized in that: The first culture tank is provided with a first pressure gauge, a first temperature sensor and a first exhaust valve outside; A second pressure gauge, a second temperature sensor and a second exhaust valve are disposed outside the second culture tank.

6. The automatic screening system for soil organic pollutant-degrading bacteria according to claim 3 is characterized in that: The colony separation module comprises: an incubator, wherein the incubator is connected to the steam generator, and a third booster pump and a third ball valve are provided between the incubator and the steam generator; a first water distributor, the first water distributor is connected to the second culture tank through a pipeline, the pipeline passes through the second rotary sealing plug and leads to the interior of the second culture tank, and a second plunger pump and a second check valve are arranged between the first water distributor and the second culture tank; A plurality of flat plates, the plurality of flat plates are circular and evenly spaced on a turntable, a third rotating shaft is provided at the center of the turntable, the third rotating shaft is connected to a motor, a fixing frame is provided on the turntable, the fixing frame is used to fix the flat plates; the flat plates are suitable for being sealed by a flat plate cover, and the flat plate cover is controlled by an electric suction cup; A dilution and coating assembly, one end of which is connected to the first culture tank through a pipeline, the pipeline passes through the third rotary sealing plug and leads to the interior of the first culture tank, a precision pump and a third check valve are arranged between the dilution and coating assembly and the first culture tank, the precision pump and the third booster pump are connected through a pipeline, and a fourth ball valve is arranged between the precision pump and the third booster pump; the other end of the dilution and coating assembly is respectively connected to a waste liquid barrel and a sterile water barrel through pipelines, a precision circulation pump and an eighth check valve are arranged between the dilution and coating assembly and the waste liquid barrel, and a fourth check valve is arranged between the waste liquid barrel and the sterile water barrel; The atomizer has a filter screen at the bottom and an atomizer nozzle at the front to make the quantitative organic pollutant liquid sprayed out in a highly dispersed state.

7. The automatic screening system for soil organic pollutant-degrading bacteria according to claim 6, characterized in that: The incubator is provided with a colony counter and a mechanical arm, wherein the colony counter is provided with a function of magnifying and imaging the colonies on the surface of the plate, so as to locate the target strain; the mechanical arm comprises a fourth rotating shaft, a rotating wheel, a first telescopic rod and a pliers, so as to select the target strain; The incubator is connected to a heating fan, the heating fan is used to adjust the temperature of the incubator, and a fifth check valve is provided between the incubator and the heating fan; The incubator is connected to a vacuum pump, and the vacuum pump is used to control the presence or absence of oxygen in the incubator. A sixth check valve is provided between the incubator and the vacuum pump; The incubator is also provided with a third pressure gauge, a third temperature sensor and a third exhaust valve.

8. The automatic screening system for soil organic pollutant-degrading bacteria according to claim 6, characterized in that: The dilution coating assembly comprises: a second telescopic rod; A mixing dish, the mixing dish being connected to the second telescopic rod via a second bolt, the mixing dish being used to mix the liquid pumped into the first culture tank and the sterile water bucket; A rotary sealing pipe, wherein the rotary sealing pipe is connected to an opening on one side of the mixing dish; A dilution coating device, the dilution coating device is connected to the rotary sealing pipe, and the bottom of the dilution coating device is provided with uniform and dense small holes; A small round rod, the small round rod is arranged below the rotary sealing pipe and is fixedly connected to the rotary sealing pipe; A hook, wherein the hook is suitable for being connected to the small round rod; A traction rope is connected to the hook, and the traction rope is connected to a fixed pulley.

9. The automatic screening system for soil organic pollutant-degrading bacteria according to claim 6, characterized in that: The colony enrichment module comprises: a telescopic pipe fitting, the telescopic pipe fitting being connected to the second check valve through a pipeline, and a seventh check valve being arranged between the telescopic pipe fitting and the second check valve; A second water distributor, the second water distributor is connected to the telescopic pipe member, and the second water distributor has a plurality of liquid outlets; A fermentation tank, wherein there are multiple fermentation tanks, and each fermentation tank is arranged below the liquid outlet; The rotating frame is provided with multiple layers, and the fermentation tank is fixed on the table of the rotating frame by a fixing clamp.

10. The automatic screening system for soil organic pollutant-degrading bacteria according to claim 9, characterized in that: Each layer of the rotating frame is provided with a plurality of independent tables; the rotating frame is controlled to rotate by a fifth rotating shaft; An electromagnet is provided at the upper end of each table surface of the rotating frame, and the electromagnet is wrapped in a sealing sleeve. Magnetic material is provided around the top of the fermentation tank cover. The size of the magnetic material circle of the fermentation tank cover is consistent with the size of the electromagnet, and the center position of the circle coincides.

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