A microplastic biofilm module culture device and its application

Abstract

The present invention belongs to the field of environmental science and technology, and discloses a microplastic biofilm module culture device and its application. In view of the various problems existing in the existing biofilm culture devices with microplastics as the matrix, a microplastic biofilm module culture device is provided, which is composed of a number of culture boxes, each of which is composed of a biofilm culture structure, a semi-automatic collection and separation structure, an aeration structure and a temperature control structure. The natural conditions required for biofilm culture are constructed and can be artificially controlled. At the same time, static and dynamic culture, aerobic, anaerobic, facultative anoxic co-culture, and temperature-controlled culture of microplastic biofilms can be realized, and relevant laboratory biofilm culture experiments under accurate simulated environmental conditions can be carried out; it can be applied to the culture of various microplastic biofilms floating, suspended or sunk in water bodies, so as to achieve the layered collection of microplastics in multiple water layers; the volume and number of biofilm culture chambers can be reasonably selected according to different needs to realize the mass production of the required biofilms.

Description

Technical Field

[0001] The present invention belongs to the field of environmental science and technology, and specifically relates to a multi-type, multi-batch, and multi-environment simulation microplastic biofilm module culture device and its application. Background Art

[0002] Microplastics are small in size, large in specific surface area, and chemically stable. They have a longer lifespan in the environment than other particles, thus providing an ideal ecological niche for microorganisms. Microplastics are light in weight and easily drift under the transport of water flow. The biofilm attached to the surface will also spread widely, and in the process affect the surrounding water environment. In recent years, with the continuous deepening of research, the complex biological communities attached to the surface of microplastics and their ecological effects have gradually attracted the attention of researchers. To study biofilms in natural water environments, it is first necessary to obtain relatively uniform biofilms. However, there are only two reports on biofilm culture devices with microplastics as the matrix in current research. The invention patent named "A biofilm culture device with microplastics as the matrix for environmental water bodies" with publication number CN 108751388 A, which places microplastics in a culture bag and connects the culture bag to a fixed plate by several scaling connecting ropes. This invention only plays the role of culturing biofilms with microplastics as the matrix, and its experimental equipment and methods have many limitations. First, the device has a single design configuration and no adjustment mechanism, which makes it difficult to simulate different culture environments, and thus it is impossible to obtain biofilms under different environmental conditions, nor to obtain multiple batches of microplastic biofilms with relatively stable properties; second, the culture bag of the device is easily impacted by underwater foreign objects, and the scaling connecting rope is easily weathered and broken, thus affecting the integrity of the biofilm morphological structure. The device is named a rapid natural water biofilm culture device, and the publication number is CN 101928059 A. The device uses a static culture method for culture, and the culture fluid in contact with the biofilm surface during the culture process cannot be quickly updated and biological metabolic waste cannot be removed, thus affecting the growth of microorganisms. Summary of the invention

[0003] The current biofilm culture devices using microplastics as the matrix mainly have the following problems:

[0004] 1. The biofilm culture device is not convenient for comparing biofilms under different culture environments, and it is impossible to obtain microplastic biofilms with relatively stable properties through existing devices.

[0005] 2. There are many types of microplastics, which appear in various states in water, such as floating, suspended, and precipitated. The current biofilm culture devices cannot solve the problem of culturing microplastic biofilms in different states, such as floating, suspended, and precipitated, with the same device.

[0006] 3. Some simple biofilm culture devices mostly use static culture methods. During the culture process, the metabolites of biofilm microorganisms are not easy to be replaced quickly, which in turn affects the growth of biofilm.

[0007] The present invention provides a multi-type, multi-batch, and multi-environment simulation microplastic biofilm module culture device and application thereof.

[0008] In order to solve the above problems, the present invention adopts the following technical solutions:

[0009] A microplastic biofilm module culture device, comprising a plurality of culture boxes, each of which comprises a biofilm culture structure, a semi-automatic collection and separation structure, an aeration structure and a temperature control structure;

[0010] The biofilm culture structure includes a biofilm culture chamber, a microplastic addition port, a microplastic addition port sealing cover, a nutrient delivery port, and a microplastic matrix; the biofilm culture chambers are independent structures, separated from each other, and the number of culture chambers can be selected according to experimental requirements, and a certain number of microplastics are placed in the biofilm culture chambers for biofilm culture experiments;

[0011] The semi-automatic collection and separation structure includes a process sampling tube, an arc-shaped collection tube baffle and a bottom microplastic discharge port; the process sampling tube is used for sampling during the experiment and collecting suspended layer samples at the end of the experiment; the arc-shaped collection tube is mainly used to collect biofilm samples in the floating layer; the bottom microplastic discharge port is mainly used to collect microplastic biofilm samples in the sedimentation layer, thereby completing the stratified collection of biofilm samples;

[0012] The aeration structure includes a gas aeration inlet, an umbrella-shaped culture fluidization air jet nozzle and an aeration pipeline, compressed gas or an air pump and other aeration components; the aeration structure provides vertical water flow to keep the system in a continuous mixing state, simulating the biofilm growth environment in natural water bodies;

[0013] The temperature control structure includes a silica gel heating plate, a temperature control sensor, and a key-type temperature digital display controller; it provides temperature control conditions for microplastic biofilm culture, and aerates the culture box to make the temperature evenly conducted;

[0014] A microplastic addition port is provided at the top of the biofilm culture chamber, and a microplastic addition port sealing cover is provided correspondingly, and the microplastic addition port sealing cover cooperates with the microplastic addition port to complete the sealing, and the nutrient material delivery port is provided on one side of the top of the biofilm culture chamber and is sealed by a rubber cap;

[0015] There are two types of sealing covers for the microplastic addition port shown, one is a single-seal sealing cover, and the other is a sampling sealing cover with a process sampling tube inserted through the center of the sealing cover;

[0016] The arc-shaped collecting cylinder as shown is similar to a hollow cylinder as a whole, and is a semi-closed arc-shaped structure. The arc surface of the cylinder is 240 / 360°, and a slide groove is provided on the vertical edge of the arc surface for installing the collecting cylinder baffle. The size of the top of the arc-shaped collecting cylinder can match the microplastic addition port, and a lifting rod A is fixedly provided for layered collection of suspended layer microplastic biofilm samples;

[0017] The collecting tube baffle is of rectangular structure. The two sides of the collecting tube baffle are matched with the slide grooves, and can be inserted into the slide grooves for sliding. A lifting rod B is fixedly provided on the top of the collecting tube baffle. The collecting tube baffle is adjusted by the lifting rod B to realize the sampling and closing of the arc-shaped collecting tube, and the collecting tube baffle cooperates with the arc-shaped collecting tube to realize the sealing of the arc-shaped collecting tube;

[0018] A bottom microplastic discharge port and two gas aeration inlets are provided at the bottom of the biofilm culture chamber; the bottom microplastic discharge port is funnel-shaped, shrinking and narrowing from the top to the bottom, and a sealing cover is provided at the lower end for sealing, so as to collect microplastic biofilm samples in the sediment layer;

[0019] The gas aeration inlet is externally connected to an aeration pipeline, and the gas aeration inlet is internally connected to an umbrella-shaped culture liquid fluidization air jet nozzle;

[0020] The silicone heating sheet is composed of two layers of insulating silicone cloth, with nickel-chromium alloy wire laid in the middle, a built-in power cord and a temperature control sensor, and a strip-shaped silicone heating sheet with adhesive backing, which is pasted on the outside of the biofilm culture chamber;

[0021] Furthermore, the sealing cover of the microplastic delivery port, the sealing cover of the bottom microplastic discharge port and the biofilm culture chamber are connected via a snap-fit ​​structure to achieve a closed state of the biofilm culture chamber.

[0022] Furthermore, a support base is provided under the microplastic biofilm module culture device. The support base is an open trapezoidal structure with a slot at the upper end. A silicone heating plate is pasted on the inside of the slot. A pipeline opening is provided on the inclined surface of one side of the trapezoid for passing an aeration pipeline and is placed under the biofilm culture chamber.

[0023] A microplastic biofilm cultivation method of a microplastic biofilm module cultivation device comprises the following steps:

[0024] Step 1, connection of the culture device: connect the biofilm culture device, connect the sealing cover of the microplastic delivery port, the sealing cover of the bottom microplastic discharge port and the biofilm culture chamber through a snap structure to achieve a closed state of the biofilm culture chamber, continue to connect the gas aeration inlet at the lower end of the biofilm culture chamber to one end of the aeration pipeline, and connect the other end of the aeration pipeline to the compressed gas or air pump through the support base, and separate the multiple culture chambers and place them in the card slot of the support base;

[0025] Step 2, culture: at the beginning of the culture, the culture chambers in the biofilm culture chamber are separated from each other, and the microplastic addition port is opened to add culture liquid, such as natural lake water, river water or homemade culture liquid, to each culture chamber, and continue to add until the liquid level reaches four-fifths of the height of the culture chamber, and add a certain number of microplastic particles; if natural water is used for culture, the culture liquid needs to be replaced at set time intervals; if a prepared solution is used for culture, a simple dosing device such as a syringe can be used to add medicine and nutrient salt solution at the nutrient addition port for easy mixing, or the prepared liquid can be used for overall replacement until the biofilm culture matures or meets the experimental requirements;

[0026] Step 3, setting and simulation of culture conditions: simulating natural water condition culture, providing aeration culture conditions, the gas aeration inlet at the lower end of the biofilm culture chamber is connected to one end of the aeration pipeline, and the other end of the aeration pipeline is connected to the compressed gas or air pump through the support base, and different culture chambers are separated from each other. Turn on the switch of the aeration component to realize the dynamic culture of the microplastic biofilm, and turn off the switch of the aeration component to realize the static culture of the microplastic biofilm; by controlling the type of gas introduced, oxygen is introduced into the aerobic biofilm culture experiment, nitrogen is introduced into the anaerobic biofilm culture experiment, and air is introduced into the facultative biofilm culture experiment, so as to realize aerobic, anaerobic, and facultative aerobic co-culture of the microplastic biofilm; provide temperature-controlled culture conditions, connect the power supply, turn on the temperature control switch on the push-button temperature digital display controller, adjust the temperature of the silicone heating plate through the temperature feedback of the temperature control sensor, or place different culture chambers in the incubator respectively, and set the temperature for culture;

[0027] Step 4, sample collection during culture: sampling during the microplastic biofilm experiment, remove the single microplastic addition port sealing cover, replace it with a sterilized microplastic addition port sealing cover with a process sampling tube, put the process sampling tube into the biofilm culture chamber, extend one end into the set sampling position, and connect the other end to the syringe, draw a certain amount of water sample and microplastic biofilm sample by pulling out the syringe after sampling, and replace the single microplastic addition port sealing cover;

[0028] Step 5, collection of floating, suspended and sinking microplastics: During the collection and separation process of microplastic biofilms, the baffle of the collection tube is pushed and pulled downward along the side slide groove of the arc-shaped collection tube to the bottom through the lifting rod B, so that the lower end of the arc-shaped collection tube is in a closed state. When in use, the arc-shaped collection tube and the baffle of the collection tube are cleaned and sterilized in advance, and placed in the biofilm culture chamber through the microplastic addition port two days in advance. When the arc-shaped collection tube sinks to the preset position in the biofilm culture chamber, the lifting rod B on the baffle of the collection tube is used to push it downward along the side slide groove of the arc-shaped collection tube until the baffle of the collection tube is just submerged in the water surface, and the upper water sample of the biofilm culture chamber and the suspended microplastic biofilm automatically flow into the arc-shaped collection tube. After the upper water sample and the floating microplastic biofilm are completely collected, the baffle of the collection tube is pulled upward along the side slide groove of the arc-shaped collection tube to close the arc-shaped collection tube. Finally, the arc-shaped collection tube is pulled upward by the lifting rod A to remove it from the biofilm culture chamber, and the sample is sucked out with a syringe to collect the floating layer water sample. Put the sterilized microplastic injection port sealing cover with process sampling tube into the microplastic injection port, put the process sampling tube into the biofilm culture chamber, and collect the suspended layer biofilm samples. Finally, open the bottom microplastic discharge port sealing cover to allow the bottom layer liquid to flow out from below to collect the sediment layer biofilm samples;

[0029] Step 6, dismantling and cleaning of the device: after the cultivation is completed, the aeration structure and the temperature control structure are closed, the biofilm samples inside the biofilm culture chamber are collected and separated by the semi-automatic collection and separation structure, the biofilm culture chamber is separated from the supporting base, and then the biofilm culture chamber is lifted upward. A gas aeration access port is provided at the bottom of the biofilm culture chamber, and the gas aeration access port and the aeration pipeline are connected to each other. As the biofilm culture chamber is lifted upward, it is convenient to separate the gas aeration access port from the aeration pipeline, and then the biofilm culture chamber is separated from the microplastic addition port and the sealing cover of the bottom microplastic discharge port. The cultivation device is separated, which is convenient for cleaning each biofilm culture component.

[0030] A microplastic biofilm module culture device is used in the culture of various microplastic biofilms that float, suspend or sink in water.

[0031] Compared with the prior art, the present invention has the following advantages:

[0032] 1. Through the cooperation of other auxiliary devices, the natural conditions required for biofilm cultivation are constructed and can be artificially controlled. By controlling the switch of the aeration component, the static and dynamic cultivation of microplastic biofilms can be achieved; by controlling the type of gas introduced, aerobic, anaerobic, and facultative aerobic co-cultivation of microplastic biofilms can be achieved; by adjusting the switch of the temperature control component, or placing different culture chambers in the incubator, the temperature-controlled cultivation of microplastic biofilms can be achieved, and relevant laboratory biofilm cultivation experiments under precise simulated environmental conditions can be carried out;

[0033] 2. It can be applied to the cultivation of various microplastic biofilms floating, suspended or sunk in water bodies, so as to achieve the stratified collection of microplastics in multiple water layers;

[0034] 3. The culture device uses flowing water to culture the biofilm, which is conducive to the timely renewal of the surface liquid of the biofilm and the growth of the biofilm;

[0035] 4. The device is designed to be partitioned, which is suitable for the separation and co-cultivation of various types of microplastics such as PP, PE, PS, and microplastic biofilms with various particle sizes;

[0036] 5. The volume and quantity of the overall culture device are adjustable. The volume and quantity of the biofilm culture chambers can be reasonably selected according to different experimental requirements to achieve mass production of the required biofilm. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 It is an overall schematic diagram of a microplastic biofilm module culture device of the present invention;

[0038] Figure 2 A schematic diagram of a culture chamber in a process sampling state of a microplastic biofilm module culture device of the present invention;

[0039] Figure 3 A schematic diagram of a collection and separation state culture chamber of a microplastic biofilm module culture device of the present invention;

[0040] Figure 4 A schematic diagram of a microplastic biofilm module culture device of the present invention with a process sampling tube sealing cover and an arc-shaped collection cylinder;

[0041] Figure 5 It is a schematic diagram of the expansion of the temperature control structure of a microplastic biofilm module culture device of the present invention.

[0042] In the figure:

[0043] 1. Biofilm culture chamber; 2. Microplastic addition port; 3. Microplastic addition port sealing cover; 4. Nutrient delivery port; 5. Microplastic matrix; 6. Process sampling tube; 7. Support base; 8. Arc-shaped collection cylinder; 801. Lifting rod A; 9. Collection cylinder baffle; 901. Lifting rod B; 10. Slide; 11. Bottom microplastic discharge port; 12. Gas aeration inlet; 13. Umbrella-shaped culture fluid fluidization air nozzle; 14. Aeration pipeline; 15. Silicone heating plate; 16. Temperature control sensor probe; 17. Push-button temperature digital display controller. DETAILED DESCRIPTION

[0044] In order to facilitate understanding of the above technical solutions of the present invention, the above technical solutions of the present invention are described in detail below through specific usage methods.

[0045] Example 1

[0046] A microplastic biofilm module culture device, comprising a plurality of culture boxes, each of which comprises a biofilm culture structure, a semi-automatic collection and separation structure, an aeration structure and a temperature control structure;

[0047] The biofilm culture structure comprises a biofilm culture chamber 1, a microplastic addition port 2, a microplastic addition port sealing cover 3, a nutrient delivery port 4, and a microplastic matrix 5;

[0048] The semi-automatic collection and separation structure includes a process sampling tube 6, an arc-shaped collection tube 8, a collection tube baffle 9 and a bottom microplastic discharge port 11;

[0049] The aeration structure includes a gas aeration inlet 12, an umbrella-shaped culture fluidization air jet nozzle 13 and an aeration pipeline 14;

[0050] The temperature control structure includes a silicone heating sheet 15, a temperature control sensor 16, and a key-type temperature digital display controller 17;

[0051] A microplastic addition port 2 is provided at the top of the biofilm culture chamber 1, and a microplastic addition port sealing cover 3 is provided correspondingly, and the microplastic addition port sealing cover 3 cooperates with the microplastic addition port 2 to complete the sealing, and the nutrient material delivery port 4 is provided on one side of the top of the biofilm culture chamber 1 and is sealed by a rubber cap;

[0052] The microplastic addition port sealing cover 3 shown is divided into two types, one is a single-seal sealing cover, and the other is a sampling sealing cover with a process sampling tube 6 passing through the center of the sealing cover;

[0053] The arc-shaped collecting cylinder 8 is similar to a hollow cylinder as a whole, and is a semi-closed arc-shaped structure. The arc surface of the cylinder is 240 / 360°, and a slide groove 10 is provided on the vertical edge of the arc surface for installing the collecting cylinder baffle 9. The top size of the arc-shaped collecting cylinder 8 can match the microplastic addition port 2, and a lifting rod A 801 is fixedly provided for layered collection of suspended layer microplastic biofilm samples;

[0054] The collecting tube baffle 9 is of rectangular structure, and the two sides of the collecting tube baffle 9 are matched with the slide grooves 10, which can be inserted into the slide grooves for sliding. A lifting rod B 901 is fixedly provided on the top of the collecting tube baffle 9, and the collecting tube baffle 9 is adjusted by the lifting rod B 901 to realize the sampling and closing of the arc-shaped collecting tube 8, and the collecting tube baffle 9 cooperates with the arc-shaped collecting tube 8 to realize the sealing of the arc-shaped collecting tube 8;

[0055] A bottom microplastic discharge port 11 and two gas aeration inlets 12 are provided at the bottom of the biofilm culture chamber 1; the bottom microplastic discharge port 11 is funnel-shaped, shrinking and narrowing from the top to the bottom, and a sealing cover is provided at the lower end for sealing, so as to collect microplastic biofilm samples in the sediment layer;

[0056] The gas aeration inlet 12 is externally connected to an aeration pipeline 14, and the gas aeration inlet 12 is internally connected to an umbrella-shaped culture liquid fluidization air jet nozzle 13;

[0057] The silicone heating sheet 15 is composed of two layers of insulating silicone cloth, with nickel-chromium alloy wire laid in the middle, and a built-in power line and a temperature control sensor 16.

[0058] Example 2 Composition, material, structure and function of microplastic biofilm module culture device

[0059] A microplastic biofilm module culture device, comprising a plurality of culture boxes, each of which comprises a biofilm culture structure, a semi-automatic collection and separation structure, an aeration structure and a temperature control structure;

[0060] The biofilm culture structure comprises a biofilm culture chamber 1, a microplastic addition port 2, a microplastic addition port sealing cover 3, and a nutrient addition port 4;

[0061] The semi-automatic collection and separation structure includes a process sampling tube 6, an arc-shaped collection tube 8, a collection tube baffle 9 and a bottom microplastic discharge port 11;

[0062] The aeration structure includes a gas aeration inlet 12, an umbrella-shaped culture fluidization air jet nozzle 13 and an aeration pipeline 14;

[0063] The temperature control structure includes a silicone heating sheet 15, a temperature control sensor 16, and a key-type temperature digital display controller 17;

[0064] The biofilm culture chamber 1 is made of glass, and is a semi-enclosed cylinder with a diameter:height ratio of 3:1. Each biofilm culture chamber 1 is separated from each other and placed in the card slot of the support base 7; the microplastic matrix 5 is placed inside the biofilm culture chamber 1 for co-cultivation experiments of floating, sinking and suspended microplastic biofilms.

[0065] The microplastic addition port 2 is made of glass and is arranged at the top of each biofilm culture chamber 1. A silicone sealing ring is placed on the edge of the microplastic addition port 2, and a microplastic addition port sealing cover 3 is placed on the upper part. The microplastic addition port sealing cover 3 is fixedly connected to the microplastic addition port 2 by a snap-fit ​​structure, and is used for adding microplastics and sampling microplastic samples.

[0066] The sealing cover 3 for the microplastic addition port is made of glass. There are two types of sealing covers 3 for the microplastic addition port, one is a single-sealing sealing cover, and the other is a sampling sealing cover with a process sampling tube 6 passing through the center of the sealing cover; the single-sealing sealing cover is used to seal the microplastic addition port 2 during the biofilm culture process, and the sampling sealing cover with a process sampling tube 6 passing through the center of the sealing cover is used to seal the microplastic addition port during the biofilm sampling process.

[0067] The nutrient delivery port 4 is made of glass and is arranged on one side of the top of the biofilm culture chamber 1. A section of a circular tube protrudes outward from the edge of the delivery port in a direction perpendicular to the culture box surface, and a sealing rubber cap is placed on the top; it is used for delivering nutrients and also serves as an air pressure balance port.

[0068] The microplastic matrix 5 is made of plastic, and the microplastic matrix is ​​a variety of microplastics such as PP, PE, PS, etc. with various particle sizes. It is added into the biofilm culture chamber 1 through the microplastic addition port 2 to provide microbial attachments and conduct biofilm culture experiments with microplastics as the matrix.

[0069] The process sampling tube 6 is made of silicone. The process sampling tube 6 is passed through the center of the microplastic addition port sealing cover 3 and can be adjusted and positioned for sampling microplastics during the cultivation process.

[0070] The support base 7 is made of acrylic, and is an open trapezoidal structure. A card slot is provided at the upper end, and a silicone heating plate 16 is pasted on the inner side of the card slot. A pipeline opening is provided on the inclined surface and is placed below the biofilm culture chamber 1. An external compressed gas or air pump is installed. The biofilm culture structure is columnar and supported by the support base. The support base has a built-in gas pipeline to save space.

[0071] The arc-shaped collecting tube 8 is made of glass and is similar to a hollow cylinder as a whole. It is a semi-closed arc-shaped structure. The arc surface of the cylinder is 240 / 360°. Slide grooves 10 are provided on the inner walls at both ends of the vertical edges of the arc surface for installing the collecting tube baffle 9. A sampling cavity is provided on the inner side of the arc-shaped collecting tube 8. The size of the top of the arc-shaped collecting tube 8 can match the microplastic addition port 2. A lifting rod A 801 is fixed on the top of the arc-shaped collecting tube 8 and is placed inside the biofilm culture chamber 1 when collecting and separating biofilm samples to perform layered collection of suspended layer microplastic biofilm samples.

[0072] The collecting tube baffle 9 is made of glass and has a rectangular structure. The two sides of the collecting tube baffle 9 are provided with slide grooves 10 and can be inserted into the slide grooves for sliding. A lifting rod B 901 is fixedly provided on the top of the collecting tube baffle 9. The collecting tube baffle 9 is adjusted by the lifting rod B 901 to realize sampling and closing of the arc-shaped collecting tube 8, and the collecting tube baffle 9 cooperates with the arc-shaped collecting tube 8 to realize the sealing of the arc-shaped collecting tube 8.

[0073] The bottom microplastic discharge port 11 is made of glass and is arranged at the bottom of the biofilm culture chamber 1. The bottom microplastic discharge port 11 is trumpet-shaped, shrinking and narrowing from the top to the bottom, and a sealing cover is provided at the lower end for collecting microplastic biofilm samples in the sedimentation layer.

[0074] The gas aeration inlet 12 is made of glass and is arranged at the bottom of the biofilm culture chamber 1. The gas aeration inlet 12 is externally connected to the aeration pipeline 14 and internally connected to the umbrella-shaped culture liquid fluidization air nozzle 13. A filter is also arranged inside the gas aeration inlet 12. Aeration is performed through the aeration ports in different biofilm culture chambers to realize static and dynamic cultivation of the biofilm. Different gases such as air, oxygen and nitrogen are provided through the gas aeration inlet to realize aerobic, anaerobic and facultative aerobic co-cultivation of the biofilm.

[0075] The umbrella-shaped culture fluidization air nozzle 13 is made of silicone. The umbrella-shaped culture fluidization air nozzle 13 is set to an elastic structure and is connected to the gas aeration inlet 12. During aeration, the airflow is uniformly fluidized inside the device, so that the microorganisms on the microplastics are fully in contact with the gas. When the aeration is turned off, it automatically closes due to the pressure relief effect.

[0076] The aeration pipeline 14 is made of silicone, one end of which is sleeved with the lower end of the gas aeration inlet 12, and then the other end of the gas pipeline is connected to the compressed gas or air pump through the support base 7; it is used to provide aeration conditions for microplastic biofilm cultivation.

[0077] The silicone heating sheet 15 is made of silicone. The silicone heating sheet 15 is divided into two layers of insulating silicone cloth, with nickel-chromium alloy wire laid in the middle, and a built-in power cord and temperature control sensor 16. The silicone heating sheet 15 has a backing adhesive and is attached to the outside of the biofilm culture chamber 1. It is used to provide temperature control conditions for microplastic biofilm culture, and the temperature is evenly conducted by aerating the culture box.

[0078] The temperature control sensor 16 has a probe placed inside the silicone heating plate 15 and a temperature measuring line connected to a key-type temperature digital display controller 17 for feeding back the temperature of the culture box for temperature regulation.

[0079] The push-button temperature digital display controller 17 is provided with a silicone heating temperature control switch, a temperature adjustment button, a temperature display screen and an automatic start-stop program. The push-button temperature digital display controller circuit is connected to one end of the silicone heating plate 15; it is used to display the real-time temperature of the culture box and automatically control the switch of the temperature control component and adjust the required temperature.

[0080] Example 2

[0081] The microplastic biofilm cultivation method of the microplastic biofilm module cultivation device as described in Example 1:

[0082] Step 1, connection of the culture device: connect the biofilm culture device, connect the sealing cover 3 of the microplastic delivery port, the sealing cover of the bottom microplastic discharge port 11 and the biofilm culture chamber 1 through a snap structure to achieve a closed state of the biofilm culture chamber, continue to connect the gas aeration inlet 12 at the lower end of the biofilm culture chamber 1 to one end of the aeration pipeline 14, and the other end of the aeration pipeline 14 is connected to the compressed gas or air pump through the support base 7, and multiple culture chambers are separated and placed in the card slot of the support base 7;

[0083] Step 2, culture: at the beginning of culture, the culture chambers in the biofilm culture chamber 1 are separated from each other, the microplastic addition port 2 is opened to add culture liquid to each culture chamber, and the addition is continued until the liquid level reaches four-fifths of the height of the culture chamber, and a certain number of microplastic particles are added; if natural water is used for culture, the culture liquid needs to be replaced at set time intervals; if a prepared solution is used for culture, the drug and nutrient salt solution can be added at the nutrient addition port 4 for easy mixing, or the prepared liquid can be used for overall replacement until the biofilm culture matures or meets the experimental requirements;

[0084] Step 3, setting and simulation of culture conditions: simulating natural water condition culture, providing aeration culture conditions, the two gas aeration inlets 12 at the lower end of the biofilm culture chamber 1 are connected to one end of the aeration pipeline 14, and the other end of the aeration pipeline 14 is connected to the compressed gas or air pump through the support base 7. Different culture chambers are separated from each other. Turn on the switch of the aeration component to realize the dynamic culture of the microplastic biofilm, and turn off the switch of the aeration component to realize the static culture of the microplastic biofilm; by controlling the type of gas introduced, oxygen is introduced into the aerobic biofilm culture experiment, nitrogen is introduced into the anaerobic biofilm culture experiment, and air is introduced into the facultative biofilm culture experiment, so as to realize aerobic, anaerobic, and facultative aerobic co-culture of the microplastic biofilm; provide temperature-controlled culture conditions, connect the power supply, turn on the temperature control switch on the push-button temperature digital display controller 17, adjust the temperature of the silica gel heating plate 15 through the temperature feedback of the temperature control sensor 16, or place different culture chambers in the incubator respectively, and set the temperature for culture;

[0085] Step 4, sample collection during culture: sampling during the microplastic biofilm experiment, taking out the single microplastic addition port sealing cover 3, replacing it with a sterilized microplastic addition port sealing cover 3 with a process sampling tube, placing the process sampling tube into the biofilm culture chamber 1, extending one end into the set sampling position, and connecting the other end to the syringe, drawing the syringe to absorb a certain amount of water sample and microplastic biofilm sample, pulling out the syringe after sampling, and replacing the single microplastic addition port sealing cover;

[0086] Step 5, collection of floating, suspended and sinking microplastics: In the process of collecting and separating microplastic biofilms, the collection tube baffle 9 is pushed and pulled downward along the side slide groove of the arc-shaped collection tube 8 to the bottom by the lifting rod B 801, so that the lower end of the arc-shaped collection tube 8 is in a closed state. When in use, the arc-shaped collection tube 8 and the collection tube baffle 9 are cleaned and sterilized in advance, and placed in the biofilm culture chamber 1 through the microplastic addition port 2 two days in advance. When the arc-shaped collection tube 8 sinks to the preset position in the biofilm culture chamber 1, the collection tube baffle 9 is lifted by the lifting rod B 901 and pushed downward along the side slide groove of the arc-shaped collection tube 8 until the collection tube baffle 9 is just submerged in the water surface. The upper water sample of the biofilm culture chamber and the suspended microplastic biofilm automatically flow into the arc-shaped collection tube 8. After the upper water sample and the floating microplastic biofilm are completely collected, the collection tube baffle 9 is pulled upward along the side slide groove of the arc-shaped collection tube 8 to close the arc-shaped collection tube 8, and finally the lifting rod A is used to collect the microplastic biofilm in the upper layer of the biofilm culture chamber. 801 Pull the arc-shaped collection tube 8 upward to remove it from the biofilm culture chamber 1, and use a syringe to suck out the sample to collect the floating layer water sample. Put the sterilized microplastic injection port sealing cover 3 with the process sampling tube into the microplastic injection port 2, and put the process sampling tube into the biofilm culture chamber 1 to collect the suspended layer biofilm sample. Finally, open the sealing cover of the bottom microplastic discharge port 11 to allow the bottom layer liquid to flow out from the bottom to collect the sedimentation layer biofilm sample;

[0087] Step 6, dismantling and cleaning of the device: after the cultivation is completed, the aeration structure and the temperature control structure are closed, the biofilm samples inside the biofilm culture chamber 1 are collected and separated by the semi-automatic collection and separation structure, the biofilm culture chamber 1 is separated from the support base 7, and then the biofilm culture chamber 1 is lifted upward. A gas aeration access port 12 is provided at the bottom of the biofilm culture chamber 1, and the gas aeration access port 12 and the aeration pipeline 14 are mutually connected. As the biofilm culture chamber 1 is lifted upward, it is convenient to separate the gas aeration access port 12 from the aeration pipeline 14, and then the biofilm culture chamber 1 is separated from the microplastic addition port 3 and the sealing cover of the bottom microplastic discharge port 11. The culture device is separated, which is convenient for cleaning each biofilm culture component.

[0088] The contents not described in detail in the specification of the present invention belong to the prior art known to the professional and technical personnel in the field. Although the illustrative specific embodiments of the present invention are described above to facilitate the understanding of the present invention by the technical personnel in the field, it should be clear that the present invention is not limited to the scope of the specific embodiments. For the ordinary technical personnel in the field, as long as various changes are within the spirit and scope of the present invention defined and determined by the attached claims, these changes are obvious, and all inventions and creations using the concept of the present invention are protected.

Claims

1. A microplastic biofilm module culture device, characterized in that: It is composed of several culture boxes, each of which is composed of a biofilm culture structure, a semi-automatic collection and separation structure, an aeration structure and a temperature control structure; The biofilm culture structure comprises a biofilm culture chamber (1), a microplastic addition port (2), a microplastic addition port sealing cover (3), a nutrient addition port (4), and a microplastic matrix (5); The semi-automatic collection and separation structure comprises a process sampling tube (6), an arc-shaped collection tube (8), a collection tube baffle (9) and a bottom microplastic discharge port (11); The aeration structure comprises a gas aeration inlet (12), an umbrella-shaped culture liquid fluidization air jet nozzle (13) and an aeration pipeline (14); The temperature control structure comprises a silicone heating plate (15), a temperature control sensor (16), and a key-operated temperature digital display controller (17); A microplastic addition port (2) is provided at the top of the biofilm culture chamber (1), and a microplastic addition port sealing cover (3) is provided correspondingly, and the microplastic addition port sealing cover (3) cooperates with the microplastic addition port (2) to complete the sealing, and the nutrient material delivery port (4) is provided at one side of the top of the biofilm culture chamber (1) and is sealed by a rubber cap; The microplastic addition port sealing cover (3) is divided into two types, one is a single-seal sealing cover, and the other is a sampling sealing cover with a process sampling tube (6) passing through the center of the sealing cover; The arc-shaped collecting cylinder (8) is a semi-enclosed arc-shaped structure as a whole, the arc surface of the cylinder is 240 / 360 degrees, and a slide groove (10) is provided on the vertical edge of the arc surface for installing the collecting cylinder baffle (9). The size of the top of the arc-shaped collecting cylinder (8) can match the microplastic addition port (2), and a lifting rod A (801) is fixedly provided on the top of the arc-shaped collecting cylinder (8) for layered collection of suspended layer microplastic biofilm samples; The collecting tube baffle (9) has a rectangular structure. Both sides of the collecting tube baffle (9) are provided with slide grooves (10) and can be inserted into the slide grooves for sliding. A lifting rod B (901) is fixedly provided on the top of the collecting tube baffle (9). The collecting tube baffle (9) is adjusted by the lifting rod B (901) to realize sampling and closing of the arc-shaped collecting tube (8). The collecting tube baffle (9) cooperates with the arc-shaped collecting tube (8) to realize the sealing of the arc-shaped collecting tube (8). A bottom microplastic discharge port (11) and two gas aeration inlets (12) are provided at the bottom of the biofilm culture chamber (1); the bottom microplastic discharge port (11) is funnel-shaped, shrinking and narrowing from the top to the bottom, and a sealing cover is provided at the lower end for sealing and for collecting microplastic biofilm samples in the sediment layer; The gas aeration inlet (12) is externally connected to an aeration pipeline (14), and the gas aeration inlet (12) is internally connected to an umbrella-shaped culture liquid fluidization air jet nozzle (13); The silicone heating sheet (15) comprises two layers of insulating silicone cloth, an upper layer and a lower layer, with a nickel-chromium alloy wire laid in the middle, and a built-in power cord and a temperature control sensor (16).

2. A microplastic biofilm module culture device according to claim 1, characterized in that: The sealing cover of the microplastic delivery port (3), the sealing cover of the bottom microplastic discharge port (11), and the biofilm culture chamber (1) are connected via a snap-fit ​​structure to achieve a closed state of the biofilm culture chamber.

3. A microplastic biofilm module culture device according to claim 1, characterized in that: A support base (7) is provided under the microplastic biofilm module culture device. The support base (7) is an open trapezoidal structure with a slot provided at the upper end. A silicone heating plate (15) is adhered to the inner side of the slot. A pipeline opening is provided on the inclined surface of one side of the trapezoid for passing an aeration pipeline (14) and is placed under the biofilm culture chamber (1).

4. The microplastic biofilm cultivation method of a microplastic biofilm module cultivation device according to claim 1, characterized in that: Step 1, connection of the culture device: connect the biofilm culture device, connect the sealing cover of the microplastic delivery port (3) and the sealing cover of the bottom microplastic discharge port (11) to the biofilm culture chamber (1) through a snap-fit ​​structure to achieve a closed state of the biofilm culture chamber, continue to connect the gas aeration inlet (12) at the lower end of the biofilm culture chamber (1) to one end of the aeration pipeline (14), and connect the other end of the aeration pipeline (14) to the compressed gas or air pump through the support base (7), and separate the multiple culture chambers and place them in the card slot of the support base (7); Step 2, culture: at the beginning of the culture, the culture chambers in the biofilm culture chamber (1) are separated from each other, the microplastic addition port (2) is opened to add culture liquid to each culture chamber, and the addition is continued until the liquid level reaches four-fifths of the height of the culture chamber, and a certain amount of microplastic particles are added; if natural water is used for culture, the culture liquid needs to be replaced at a set time interval; if a prepared solution is used for culture, the drug and nutrient salt solution can be added at the nutrient addition port (4) for easy mixing, or the prepared liquid can be used for overall replacement until the biofilm culture matures or meets the experimental requirements; Step 3, setting and simulating culture conditions: simulating culture under natural water conditions, providing aeration culture conditions, two gas aeration inlets (12) at the lower end of the biofilm culture chamber (1) are connected to one end of the aeration pipeline (14), and the other end of the aeration pipeline (14) is connected to compressed gas or an air pump through a support base (7). Different culture chambers are separated from each other. Turning on the switch of the aeration component realizes dynamic culture of microplastic biofilms, and turning off the switch of the aeration component realizes static culture of microplastic biofilms. By controlling the type of gas introduced, oxygen is introduced in aerobic biofilm culture experiments, nitrogen is introduced in anaerobic biofilm culture experiments, and air is introduced in facultative biofilm culture experiments, so as to realize aerobic, anaerobic, and facultative co-culture of microplastic biofilms. Providing temperature-controlled culture conditions, connecting a power supply, turning on the temperature control switch on the push-button temperature digital display controller (17), and adjusting the temperature of the silicone heating plate (15) through temperature feedback from the temperature control sensor (16), or placing different culture chambers in an incubator and setting the temperature for culture. Step 4, collecting samples during culture: sampling during the microplastic biofilm experiment, taking out the single microplastic addition port sealing cover (3), and replacing it with a sterilized microplastic addition port sealing cover (3) with a process sampling tube, placing the process sampling tube into the biofilm culture chamber (1), extending one end into the set sampling position, and connecting the other end to a syringe, drawing the syringe to absorb a certain amount of water sample and microplastic biofilm sample, pulling out the syringe after sampling, and replacing the single microplastic addition port sealing cover; Step 5, collection of floating, suspended and sinking microplastics: During the process of collecting and separating microplastic biofilms, the collection tube baffle (9) is pushed and pulled downward along the side slide groove of the arc-shaped collection tube (8) to the bottom by the lifting rod B (901), so that the lower end of the arc-shaped collection tube (8) is in a closed state. When in use, the arc-shaped collection tube (8) and the collection tube baffle (9) are cleaned and sterilized in advance, and are placed in the biofilm culture chamber (1) through the microplastic addition port (2) two days in advance. When the arc-shaped collection tube (8) sinks to a preset position in the biofilm culture chamber (1), the collection tube baffle (9) is lifted by the lifting rod B (901) ) push it downward along the side slide groove of the arc-shaped collecting tube (8) until the collecting tube baffle (9) just submerges into the water surface, and the upper water sample of the biofilm culture box and the microplastic biofilm in a suspended state automatically flow into the arc-shaped collecting tube (8). After the upper water sample and the microplastic biofilm in a floating state are completely collected, pull the collecting tube baffle (9) upward along the side slide groove of the arc-shaped collecting tube (8) to close the arc-shaped collecting tube (8). Finally, pull the arc-shaped collecting tube (8) upward by using the lifting rod A (801) to remove it from the biofilm culture chamber (1), and use a syringe to suck out the sample to collect the floating layer water sample; A sterilized microplastic injection port sealing cover (3) with a process sampling tube is placed from the microplastic injection port (2), and the process sampling tube is placed in the biofilm culture chamber (1) to collect suspended layer biofilm samples; finally, the sealing cover of the bottom microplastic discharge port (11) is opened to allow the bottom layer liquid to flow out from below, thereby collecting the sediment layer biofilm samples; Step 6, device dismantling and cleaning: After the cultivation is completed, the aeration structure and the temperature control structure are closed, and the biofilm sample inside the biofilm culture chamber (1) is collected and separated by the semi-automatic collection and separation structure, and the biofilm culture chamber (1) is separated from the support base (7), and then the biofilm culture chamber (1) is lifted upward. A gas aeration access port (12) is provided at the bottom of the biofilm culture chamber (1), and the gas aeration access port (12) and the aeration pipeline (14) are mutually connected. As the biofilm culture chamber (1) is lifted upward, it is convenient to separate the gas aeration access port (12) from the aeration pipeline (14), and then the biofilm culture chamber (1) is separated from the microplastic addition port (3) and the sealing cover of the bottom microplastic discharge port (11). The cultivation device is separated, and it is convenient to clean each biofilm culture component.

5. An application of the microplastic biofilm module culture device as claimed in claim 1 in the culture of various microplastic biofilms floating, suspended or sunk in water.

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