CN115478008A - Microplastic biofilm module culture device and application thereof - Google Patents

Microplastic biofilm module culture device and application thereof Download PDF

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Publication number
CN115478008A
CN115478008A CN202211177211.3A CN202211177211A CN115478008A CN 115478008 A CN115478008 A CN 115478008A CN 202211177211 A CN202211177211 A CN 202211177211A CN 115478008 A CN115478008 A CN 115478008A
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culture
micro
plastic
biomembrane
collecting cylinder
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朱宇恩
李华
闫秀荣
闫冠玉
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Shanxi University
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Abstract

The invention belongs to the technical field of environmental science and discloses a culture device for a micro plastic biofilm module and application thereof. Aiming at the problems of the prior biomembrane culture device taking the micro-plastic as the matrix, the utility model provides a module culture device of the micro-plastic biomembrane, which consists of a plurality of culture boxes, wherein each culture box consists of a biomembrane culture structure, a semi-automatic collection and separation structure, an aeration structure and a temperature control structure. Natural conditions required by biofilm culture are built, artificial control can be performed, static and dynamic culture, aerobic, anaerobic and facultative aerobic co-culture and temperature control culture of the micro plastic biofilm can be realized, and a relevant laboratory biofilm culture experiment under an accurate simulation environment condition is developed; can be applied to the culture of various micro plastic biofilms floating, suspending or sinking in water bodies to achieve the layered collection of the micro plastic on a multi-water layer; the volume and the number of the biomembrane culture chambers can be reasonably selected according to different requirements, and the batch production of the required biomembranes is realized.

Description

Microplastic biofilm module culture device and application thereof
Technical Field
The invention belongs to the technical field of environmental science, and particularly relates to a multi-type multi-batch multi-environment simulation micro-plastic biofilm module culture device and application thereof.
Background
The micro plastic has small volume, large specific surface area and stable chemical property, has longer service life in the environment compared with other particles, thus providing an ideal ecological niche for microorganisms, has light weight, is easy to drift under the transportation of water flow, and can widely spread a biofilm attached on the surface and influence the surrounding water environment in the process. In recent years, with the progress of research, researchers have attracted attention to complex biological communities growing on the surface of micro-plastics and ecological effects thereof, and in order to research a biofilm in a natural water environment, it is first required to obtain a biofilm with relatively uniform properties. The invention discloses a biological membrane culture device which is used in environmental water and takes micro-plastic as a substrate, and the invention patent with publication number CN 108751388A, the device places the micro-plastic in a culture bag, and connects the culture bag to a fixed disc by a plurality of scaling connecting ropes. The invention only plays a role of culturing the biological membrane by taking the micro plastic as a substrate, and the experimental equipment and the method have a plurality of limitations. Firstly, the device has a single design configuration, does not have an adjusting mechanism, is not convenient for simulating different culture environments, and further cannot obtain a biological membrane under different environmental conditions or obtain a micro-plastic biological membrane with relatively stable properties in multiple batches; secondly, the culture bag of the device is easily impacted by underwater foreign matters, and the scaling connecting rope is easily weathered and broken, so that the integrity of the morphological structure of the biological membrane is influenced. The device is characterized in that the device is a rapid natural water body biological membrane culture device, the publication number is CN 101928059A, the device is cultured in a static culture mode, and a culture solution which is contacted with the surface of a biological membrane in the culture process can not be rapidly updated and biological metabolic waste can not be removed, so that the growth of microorganisms is influenced.
Disclosure of Invention
The device mainly aims at the following problems of the existing biomembrane culture device taking micro plastic as a matrix:
1. the biofilm culturing device is inconvenient for comparing biofilms in different culturing environments, and a micro plastic biofilm with relatively stable properties cannot be obtained through the existing device.
2. The existing biomembrane culture device cannot solve the problem of culture of the plastic biomembranes in different states of floating, suspending and precipitating by the same device.
3. Some simple and easy biomembrane culture apparatus use static culture mode to cultivate more, and the difficult quick alternate change of biomembrane microorganism metabolite in the culture process, and then influence the biomembrane and grow.
The invention provides a multi-type multi-batch multi-environment simulated culture device for a micro plastic biofilm module and application thereof.
In order to solve the problems, the invention adopts the following technical scheme:
a micro plastic biofilm module culture device comprises a plurality of culture boxes, wherein each culture box comprises a biofilm culture structure, a semi-automatic collection and separation structure, an aeration structure and a temperature control structure;
the biological membrane culture structure comprises a biological membrane culture chamber, a micro-plastic adding port sealing cover, a nutrient substance feeding port and a micro-plastic substrate; the biofilm culturing chambers are independent structures and separated from each other, the number of the biofilm culturing chambers can be automatically selected according to the experiment requirement, and a certain amount of micro-plastic is put into the biofilm culturing chambers to carry out a biofilm culturing experiment;
the semi-automatic collecting and separating structure comprises a process sampling tube, an arc-shaped collecting cylinder baffle plate and a bottom micro-plastic discharge port; the process sampling tube is used for sampling in the experimental process and collecting a suspension layer sample when the experiment is finished; the arc-shaped collecting cylinder is mainly used for collecting a biological membrane sample of the floating layer; the bottom micro plastic discharge port is mainly used for collecting a micro plastic biofilm sample of the sediment layer so as to complete the layered collection of the biofilm sample;
the aeration structure comprises aeration components such as a gas aeration access port, an umbrella-shaped culture solution fluidization air nozzle, an aeration pipeline, a compressed gas or gas pump and the like; the aeration structure provides water flow in the vertical direction, so that the system is in a continuous mixed state, and the growth environment of a biological film in a natural water body is simulated;
the temperature control structure comprises a silica gel heating sheet, a temperature control sensor and a key-type temperature digital display controller; providing a temperature control condition for culturing the micro plastic biofilm, and aerating the culture box to uniformly conduct the temperature;
the top of the biological membrane culture chamber is provided with a micro-plastic adding port, a micro-plastic adding port sealing cover is correspondingly arranged, the micro-plastic adding port sealing cover is matched with the micro-plastic adding port to complete sealing, and the nutrient substance feeding port is arranged on one side of the top of the biological membrane culture chamber and is sealed through a rubber cap;
the sealing covers of the micro-plastic adding ports are divided into two types, one type is a single sealing type sealing cover, and the other type is a sampling sealing cover with a sampling tube arranged in the center of the sealing cover in a penetrating mode;
the arc-shaped collecting cylinder is similar to a hollow cylinder in whole and is of a semi-closed arc structure, the arc surface of the cylinder is 240/360 degrees, a sliding groove is formed in the vertical edge of the arc surface and used for installing a collecting cylinder baffle, the size of the cylinder top of the arc-shaped collecting cylinder can be matched with a micro-plastic adding port, and a lifting rod A is fixedly arranged and used for carrying out layered collection on a suspended layer micro-plastic biofilm sample;
the collecting cylinder baffle is of a rectangular structure, two sides of the collecting cylinder baffle are matched with the sliding grooves and can be inserted into the sliding grooves to slide, a lifting rod B is fixedly arranged at the top of the collecting cylinder baffle, the collecting cylinder baffle is adjusted through the lifting rod B to achieve sampling and closing of the arc-shaped collecting cylinder, and the collecting cylinder baffle is matched with the arc-shaped collecting cylinder to achieve sealing of the arc-shaped collecting cylinder;
the bottom of the biomembrane culture chamber is provided with a bottom micro-plastic discharge port and two gas aeration access ports; the bottom micro plastic discharge port is funnel-shaped, continuously shrinks and narrows from the top to the bottom, and the lower end of the bottom micro plastic discharge port is provided with a sealing cover for sealing and is used for collecting a micro plastic biofilm sample on a sediment layer;
the gas aeration access is externally connected with an aeration pipeline and internally connected with an umbrella-shaped culture solution fluidizing air nozzle;
the strip-shaped silica gel heating sheet is provided with gum and is adhered to the outside of the biomembrane culture chamber;
furthermore, the sealing cover of the micro-plastic feeding port and the sealing cover of the bottom micro-plastic discharge port are connected with the biological film culture chamber through a buckle structure so as to achieve the closed state of the biological film culture chamber.
Further, the support base is arranged below the culture device of the micro plastic biofilm module, the support base is of an open type trapezoid structure, a clamping groove is formed in the upper end of the support base, a silica gel heating sheet is pasted on the inner side of the clamping groove, and a pipeline opening is formed in an inclined plane on one side of the trapezoid and used for penetrating through an aeration pipeline and being arranged below the biofilm culture chamber.
A method for culturing a micro plastic biofilm of a micro plastic biofilm module culture device comprises the following steps:
step 1, connection of a culture device: connecting a biological membrane culture device, connecting a micro-plastic feeding port sealing cover and a bottom micro-plastic discharge port sealing cover with a biological membrane culture chamber through a buckle structure so as to achieve a closed state of the biological membrane culture chamber, continuously connecting a gas aeration inlet at the lower end of the biological membrane culture chamber to be communicated with one end of an aeration pipeline, connecting the other end of the aeration pipeline to a compressed gas or a gas pump through a support base, and separately placing a plurality of culture chambers in support base clamping grooves;
step 2, culturing: when the culture starts, the culture chambers in the biomembrane culture chamber are mutually separated, a micro-plastic adding port is opened to add culture liquid, such as natural lake water, river water or self-made culture liquid, into each culture chamber, the culture liquid is continuously added until the liquid level reaches four fifths of the height of the culture chamber, and a certain amount of micro-plastic particles are added; if a natural water body is used for culturing, the culture liquid needs to be replaced at set time intervals; if the prepared solution is used for culturing, simple medicine adding devices such as an injector and the like can be used for adding medicine and nutrient salt solution at the nutrient substance feeding port, so that the medicine and the nutrient salt solution are convenient to mix, and the prepared solution can be used for overall replacement until the biological membrane is cultured to be mature or the experimental requirements are met;
step 3, setting and simulating culture conditions: simulating the culture of natural water body conditions, providing aeration culture conditions, wherein the lower end of a biofilm culture chamber is communicated with one end of an aeration pipeline through an air aeration access port, the other end of the aeration pipeline is connected to a compressed air or an air pump through a supporting base, different culture chambers are mutually separated, a switch of an aeration component is turned on to realize the dynamic culture of the micro plastic biofilm, and the switch of the aeration component is turned off to realize the static culture of the micro plastic biofilm; by controlling the type of the gas introduced, oxygen is introduced into an aerobic biomembrane culture experiment, nitrogen is introduced into an anaerobic biomembrane culture experiment, and air is introduced into a facultative biomembrane culture experiment, so that aerobic, anaerobic and facultative co-culture of the micro-plastic biomembrane is realized; providing temperature control culture conditions, connecting a power supply, turning on a temperature control switch on a key-type temperature digital display controller, and carrying out temperature regulation on a silica gel heating sheet through temperature feedback of a temperature control sensor, or respectively placing different culture chambers in an incubator and setting temperature for culture;
step 4, collecting samples in culture: sampling in the experimental process of the micro plastic biofilm, taking out a single micro plastic feeding port sealing cover, replacing the single micro plastic feeding port sealing cover with a sterilized micro plastic feeding port sealing cover with a process sampling tube, putting the process sampling tube into a biofilm culturing chamber, extending one end of the process sampling tube into a set sampling position, connecting the other end of the process sampling tube with an injector, drawing the injector to absorb a certain amount of water sample and the micro plastic biofilm sample, pulling out the injector after sampling is finished, and replacing the single micro plastic feeding port sealing cover;
step 5, collecting the suspension and sinking tri-state micro-plastic: the collection and separation process of the micro-plastic biomembrane comprises the steps of pushing and pulling a collecting cylinder baffle plate downwards to the bottommost end along an arc-shaped collecting cylinder side sliding groove through a lifting rod B, enabling the lower end of the arc-shaped collecting cylinder to be in a closed state, cleaning and sterilizing the arc-shaped collecting cylinder baffle plate in advance when the collection device is used, putting the collection device into a biomembrane culture chamber through a micro-plastic adding port in advance, pushing the collection device downwards along the arc-shaped collecting cylinder side sliding groove through the lifting rod B of the collecting cylinder baffle plate when the arc-shaped collecting cylinder sinks to a preset position in the biomembrane culture chamber, just submerging the collection device baffle plate into the water surface, automatically flowing an upper water sample of the biomembrane culture chamber and a suspended micro-plastic biomembrane into the arc-shaped collecting cylinder, pulling the collection cylinder baffle plate upwards along the arc-shaped collecting cylinder side sliding groove after the upper water sample and the suspended micro-plastic biomembrane are completely collected, closing the arc-shaped collecting cylinder, finally pulling the arc-shaped collecting cylinder upwards through the lifting rod A, taking the collection cylinder out of the biomembrane culture chamber, sucking the sample by using an injector, and realizing the collection of the floating layer. And placing the sterilized micro plastic feeding port sealing cover with the process sampling tube from the micro plastic feeding port, and placing the process sampling tube into the biomembrane culture chamber to collect the suspended layer biomembrane sample. Finally, opening a sealing cover of a bottom micro-plastic discharge port to enable bottom liquid to flow out from the lower part, so as to realize collection of a sediment layer biofilm sample;
step 6, dismantling and cleaning the device: after the cultivation, close aeration structure and accuse temperature structure, collect the inside biomembrane sample collection separation of isolation structure with the biomembrane culture room through semi-automatization, cultivate the biomembrane room with supporting the base separation, then upwards promote the biomembrane culture room, biomembrane culture room bottom is equipped with gaseous aeration and inserts the mouth, and gaseous aeration inserts the mouth and cup joint each other with the aeration pipeline, along with the upwards promotion of biomembrane culture room, be convenient for insert the separation of mouth and aeration pipeline to gaseous aeration, carry out the separation that biomembrane culture room and little plastics add the sealed lid of mouth and bottom little plastics discharge port afterwards, culture apparatus separates promptly and accomplishes, be convenient for to the washing of each biomembrane culture assembly.
An application of a culture device of a micro plastic biomembrane module in the culture of various micro plastic biomembranes which float, suspend and sink in water.
Compared with the prior art, the invention has the following advantages:
1. through the cooperation of other auxiliary devices, natural conditions required by the culture of the biological membrane are constructed, manual control can be performed, and the static and dynamic culture of the micro plastic biological membrane is realized by controlling and adjusting the switch of the aeration component; the aerobic, anaerobic and facultative co-culture of the micro plastic biomembrane is realized by controlling the type of the introduced gas; the temperature-controlled culture of the micro plastic biofilm is realized by adjusting the switch of the temperature-controlled assembly or placing different culture rooms in an incubator, and a biofilm culture experiment of a related laboratory under the condition of accurately simulating the environment is carried out;
2. can be applied to the culture of various micro plastic biofilms floating, suspending or sinking in water body, and achieves the layered collection of the micro plastic on a multi-water layer;
3. the culture device adopts the flowing water body to culture the biological membrane, thereby being beneficial to timely updating the liquid on the surface layer of the biological membrane and being beneficial to the growth of the biological membrane;
4. the device is designed in a separation mode, and is suitable for separation and co-culture of various types of micro plastics such as PP, PE, PS and the like and micro plastic biofilms with various particle sizes;
5. the volume and the number of the whole culture device can be adjusted, and the volume and the number of the biomembrane culture chambers can be reasonably selected according to different experimental requirements, so that the batch production of the required biomembranes is realized.
Drawings
FIG. 1 is a schematic view of the whole culture apparatus of a module of a micro plastic biofilm according to the present invention;
FIG. 2 is a schematic view of a process sampling state culture chamber of a culture apparatus for a micro plastic biofilm module according to the present invention;
FIG. 3 is a schematic view of a collection and separation state culture chamber of a culture apparatus for a micro plastic biofilm module according to the present invention;
FIG. 4 is a schematic view of a process coupon seal cap and an arcuate collection canister of a microbial biofilm module culture apparatus of the present invention;
FIG. 5 is a development view of a temperature control structure of a culture apparatus for a micro plastic biofilm module according to the present invention.
In the figure:
1. a biofilm culturing chamber; 2. a micro-plastic addition port; 3. a micro plastic adding port sealing cover; 4. a nutrient substance feeding port; 5. a micro-plastic matrix; 6. a process sampling tube; 7. a support base; 8. an arc-shaped collecting cylinder; 801. lifting a pull rod A; 9. a collecting cylinder baffle; 901. lifting a pull rod B; 10. a chute; 11. a bottom micro plastic discharge port; 12. a gas aeration access port; 13. an umbrella-shaped culture fluid fluidization air nozzle; 14. an aeration pipeline; 15. a silica gel heating plate; 16. a temperature control sensor probe; 17. a push-button temperature digital display controller.
Detailed Description
For the convenience of understanding the above technical aspects of the present invention, the following detailed description will be given of the above technical aspects of the present invention in terms of specific modes of use.
Example 1
A micro plastic biofilm module culture device comprises a plurality of culture boxes, wherein each culture box comprises a biofilm culture structure, a semi-automatic collection and separation structure, an aeration structure and a temperature control structure;
the biological membrane culture structure comprises a biological membrane culture chamber 1, a micro-plastic adding port 2, a micro-plastic adding port sealing cover 3, a nutrient substance feeding port 4 and a micro-plastic substrate 5;
the semi-automatic collecting and separating structure comprises a process sampling tube 6, an arc-shaped collecting cylinder 8, a collecting cylinder baffle plate 9 and a bottom micro-plastic discharge port 11;
the aeration structure comprises a gas aeration inlet 12, an umbrella-shaped culture solution fluidization air nozzle 13 and an aeration pipeline 14;
the temperature control structure comprises a silica gel heating sheet 15, a temperature control sensor 16 and a key-type temperature digital display controller 17;
the top of the biomembrane culture chamber 1 is provided with a micro-plastic adding port 2, a micro-plastic adding port sealing cover 3 is correspondingly arranged, the micro-plastic adding port sealing cover 3 is matched with the micro-plastic adding port 2 to complete sealing, and the nutrient substance feeding port 4 is arranged on one side of the top of the biomembrane culture chamber 1 and is sealed by a rubber cap;
the sealing covers 3 of the micro-plastic adding openings are divided into two types, one type is a single sealing type sealing cover, and the other type is a sampling sealing cover with a sampling pipe 6 arranged in the center of the sealing cover in a penetrating way;
the arc-shaped collecting cylinder 8 is similar to a hollow cylinder as a whole and is of a semi-closed arc structure, the arc surface of the cylinder is 240/360 degrees, a sliding groove 10 is formed in the vertical edge of the arc surface and used for installing a collecting cylinder baffle 9, the size of the cylinder top of the arc-shaped collecting cylinder 8 can be matched with the micro-plastic adding port 2, and a lifting rod A801 is fixedly arranged and used for conducting layered collection of suspended layer micro-plastic biofilm samples;
the collecting cylinder baffle 9 is of a rectangular structure, two sides of the collecting cylinder baffle 9 are matched with the sliding grooves 10 and can be inserted into the sliding grooves to slide, a lifting rod B901 is fixedly arranged at the top of the collecting cylinder baffle 9, the collecting cylinder baffle 9 is adjusted through the lifting rod B901 to achieve sampling and closing of the arc-shaped collecting cylinder 8, and the collecting cylinder baffle 9 is matched with the arc-shaped collecting cylinder 8 to achieve sealing of the arc-shaped collecting cylinder 8;
the bottom of the biomembrane culture chamber 1 is provided with a bottom micro plastic discharge port 11 and two gas aeration access ports 12; the bottom micro plastic discharge port 11 is funnel-shaped, continuously shrinks and narrows from the top to the bottom, and the lower end is provided with a sealing cover for sealing and is used for collecting a micro plastic biofilm sample on a sediment layer;
the gas aeration inlet 12 is externally connected with an aeration pipeline 14, and an umbrella-shaped culture solution fluidization air nozzle 13 is connected in the gas aeration inlet 12;
the silica gel heating sheet 15 is an upper layer and a lower layer of insulating silica gel cloth, a nichrome wire is laid in the middle of the silica gel heating sheet, and a power line and a temperature control sensor 16 are arranged in the silica gel heating sheet.
Example 2 composition, material, structure and function of Microplastic biofilm Module culturing apparatus
A micro plastic biofilm module culture device comprises a plurality of culture boxes, wherein each culture box comprises a biofilm culture structure, a semi-automatic collection and separation structure, an aeration structure and a temperature control structure;
the biological membrane culture structure comprises a biological membrane culture chamber 1, a micro-plastic adding port 2, a micro-plastic adding port sealing cover 3 and a nutrient substance feeding port 4;
the semi-automatic collecting and separating structure comprises a process sampling tube 6, an arc-shaped collecting cylinder 8, a collecting cylinder baffle 9 and a bottom micro-plastic discharge port 11;
the aeration structure comprises a gas aeration inlet 12, an umbrella-shaped culture solution fluidization air nozzle 13 and an aeration pipeline 14;
the temperature control structure comprises a silica gel heating sheet 15, a temperature control sensor 16 and a key-type temperature digital display controller 17;
the biomembrane culture chamber 1 is made of glass and has the diameter: a semi-closed cylinder with the height =3, wherein the biomembrane culture chambers 1 are arranged in the clamping grooves of the supporting base 7 in a mutually separated mode; the interior of the biomembrane culture chamber 1 is filled with a micro-plastic matrix 5 for carrying out a floating and sinking micro-plastic biomembrane co-culture experiment.
The micro-plastic adding port 2 is made of glass and arranged at the top of each biomembrane culture chamber 1, a silica gel sealing ring is arranged at the edge of the micro-plastic adding port 2, a micro-plastic adding port sealing cover 3 is arranged on the upper part of the micro-plastic adding port, and the micro-plastic adding port sealing cover 3 and the micro-plastic adding port 2 are fixedly connected through a buckle structure and are used for adding micro-plastic and sampling micro-plastic samples.
The micro plastic adding port sealing cover 3 is made of glass, and the micro plastic adding port sealing cover 3 is divided into two types, one type is a single sealing type sealing cover, and the other type is a sampling sealing cover with a sampling tube 6 in the process of penetrating through the center of the sealing cover; the single sealed sealing cover is used for sealing the micro plastic adding port 2 in the biomembrane culture process, and the sampling sealing cover with the sampling tube 6 penetrating through the center of the sealing cover is used for sealing the micro plastic adding port in the biomembrane sampling process.
The nutrient substance feeding port 4 is made of glass, the nutrient substance feeding port 4 is arranged on one side of the top of the biological membrane culture chamber 1, a section of circular tube protrudes outwards from the edge of the feeding port along the direction vertical to the surface of the culture box, and a sealing rubber cap is arranged at the upper part of the feeding port; is used for feeding nutrient substances and also serves as an air pressure balancing port.
The micro plastic matrix 5 is made of plastic, is various micro plastics with various particle sizes such as PP, PE, PS and the like, is put into the biomembrane culture chamber 1 through the micro plastic adding port 2 and is used for providing microorganism attachment and carrying out a biomembrane culture experiment taking the micro plastic as the matrix.
The process sampling tube 6 is made of silica gel, and the process sampling tube 6 is arranged at the center of the sealing cover 3 of the micro-plastic adding port in a penetrating way and can be adjusted and positioned for sampling the micro-plastic in the culture process.
The supporting base 7 is made of acrylic, the supporting base 7 is of an open type trapezoid structure, a clamping groove is formed in the upper end of the supporting base, a silica gel heating sheet 16 is pasted on the inner side of the clamping groove, a pipeline opening is formed in the inclined surface of the supporting base and placed below the biological membrane culture chamber 1, a compressed gas or air pump is arranged outside the supporting base, the biological membrane culture structure is of a column type and is supported through the supporting base; the gas pipeline is arranged in the supporting base, so that the space is saved.
Arc-shaped collecting cylinder 8 adopts glass to make, wholly be similar to hollow cylinder, for semi-enclosed arc structure, the cylinder cambered surface is 240/360, spout 10 has been seted up on the inside wall of cambered surface vertical edge both ends, be used for installing collecting cylinder baffle 9, a sample chamber has been seted up to arc-shaped collecting cylinder 8 inboard, arc-shaped collecting cylinder 8's bobbin deck size can add mouthful 2 cooperatees with little plastics, arc-shaped collecting cylinder 8's bobbin deck is fixed to be set up and to carry pull rod A801 and place the inside of biomembrane culture chamber 1 in when collecting the separation biomembrane sample, carry out the layering of suspension layer little plastics biomembrane sample and collect.
The collecting cylinder baffle 9 is made of glass and is of a rectangular structure, two sides of the collecting cylinder baffle 9 are matched with the sliding grooves 10 and can be inserted into the sliding grooves to slide, a lifting rod B901 is fixedly arranged at the top of the collecting cylinder baffle 9, the collecting cylinder baffle 9 is adjusted through the lifting rod B901 to achieve sampling and closing of the arc-shaped collecting cylinder 8, and the collecting cylinder baffle 9 is matched with the arc-shaped collecting cylinder 8 to achieve sealing of the arc-shaped collecting cylinder 8;
the bottom micro plastic discharge port 11 is made of glass, the bottom micro plastic discharge port 11 is arranged at the bottom of the biomembrane culture chamber 1, the bottom micro plastic discharge port is horn-shaped and is continuously contracted and narrowed from the top to the bottom, and the lower end of the bottom micro plastic discharge port is provided with a sealing cover for collecting a sediment layer micro plastic biomembrane sample.
The gas aeration inlet 12 is made of glass and is arranged at the bottom of the biomembrane culture chamber 1, the gas aeration inlet 12 is externally connected with an aeration pipeline 14 and internally connected with an umbrella-shaped culture fluid fluidization air nozzle 13; a filter screen is also arranged in the gas aeration inlet 12; aeration is carried out through aeration ports in different biomembrane culture chambers to realize static and dynamic culture of the biomembrane; different gases such as air, oxygen and nitrogen are provided through the gas aeration access port, so that aerobic-anaerobic-aerobic co-culture of the biomembrane is realized.
Umbrella-shaped culture solution fluidization air nozzle 13 is made of silica gel, umbrella-shaped culture solution fluidization air nozzle 13 is of an elastic structure and is connected with gas aeration access port 12, flow equalization of air flow inside the device is achieved during aeration, microorganisms on the micro plastic are in full contact with gas, and the micro plastic is automatically closed due to pressure relief effect when aeration is closed.
The aeration pipeline 14 is made of silica gel, one end of the aeration pipeline is sleeved with the lower end of the gas aeration inlet 12, and then the other end of the gas pipeline is combined and connected to a compressed gas or a gas pump through the supporting base 7; used for providing aeration conditions for culturing the micro plastic biofilm.
The silica gel heating sheet 15 is made of silica gel, the silica gel heating sheet 15 is divided into an upper layer and a lower layer of insulating silica gel cloth, a nichrome wire is laid in the middle, and a power line and a temperature control sensor 16 are arranged in the silica gel heating sheet. The silica gel heating plate 15 is provided with gum and is stuck to the outer side of the biomembrane culture chamber 1. Is used for providing temperature control conditions for culturing the micro plastic biofilm, and leads the temperature to be uniformly conducted by aerating the culture box.
A temperature control sensor 16, wherein a probe of the temperature control sensor 16 is arranged in the silica gel heating sheet 15, and a temperature measuring line of the temperature control sensor is connected into the key-type temperature digital display controller 17; used for feeding back the temperature of the culture box to regulate and control the temperature.
The key-type temperature digital display controller 17 is provided with a silica gel heating temperature control switch, a temperature adjusting key, a temperature display screen and an automatic start-stop program, and a circuit of the key-type temperature digital display controller is connected with one end of the silica gel heating sheet 15; the temperature control device is used for displaying the real-time temperature of the culture box, automatically controlling the switch of the temperature control assembly and adjusting the required temperature.
Example 2
The method for culturing the micro plastic biofilm of the micro plastic biofilm module culture device in the embodiment 1 comprises the following steps:
step 1, connection of a culture device: connecting a biological membrane culture device, connecting a sealing cover 3 of a micro plastic feeding port and a sealing cover of a bottom micro plastic discharge port 11 with a biological membrane culture chamber 1 through a buckle structure so as to achieve a closed state of the biological membrane culture chamber, continuously connecting a gas aeration inlet 12 at the lower end of the biological membrane culture chamber 1 to be communicated with one end of an aeration pipeline 14, connecting the other end of the aeration pipeline 14 to a compressed gas or gas pump through a support base 7, and separately placing a plurality of culture chambers in clamping grooves of the support base 7;
step 2, culturing: when the culture starts, the culture chambers in the biomembrane culture chamber 1 are mutually separated, the micro-plastic adding port 2 is opened to add culture liquid into each culture chamber, and the culture liquid is continuously added until the liquid level reaches four fifths of the height of the culture chambers, and a certain amount of micro-plastic particles are added; if a natural water body is used for culturing, the culture liquid needs to be replaced at set time intervals; if the prepared solution is used for culturing, the nutrient substance feeding port 4 can be added with medicine and nutrient salt solution for mixing, or the prepared solution can be used for overall replacement until the biological membrane is cultured to be mature or the experimental requirements are met;
step 3, setting and simulating culture conditions: simulating the culture of natural water conditions, providing aeration culture conditions, wherein 2 gas aeration inlets 12 at the lower end of a biofilm culture chamber 1 are communicated with one end of an aeration pipeline 14, the other end of the aeration pipeline 14 is connected to a compressed gas or an air pump through a supporting base 7, different culture chambers are mutually separated, a switch of an aeration component is turned on to realize the dynamic culture of the micro plastic biofilm, and the switch of the aeration component is turned off to realize the static culture of the micro plastic biofilm; by controlling the type of the gas introduced, oxygen is introduced into an aerobic biomembrane culture experiment, nitrogen is introduced into an anaerobic biomembrane culture experiment, and air is introduced into a facultative biomembrane culture experiment, so that aerobic, anaerobic and facultative co-culture of the micro-plastic biomembrane is realized; providing temperature control culture conditions, connecting a power supply, turning on a temperature control switch on a key-type temperature digital display controller 17, and carrying out temperature regulation on the silica gel heating sheet 15 through temperature feedback of a temperature control sensor 16, or respectively placing different culture chambers in an incubator and setting temperature for culture;
step 4, collecting samples in culture: sampling in the experimental process of the micro plastic biofilm, taking out the single micro plastic feeding port sealing cover 3, replacing the single micro plastic feeding port sealing cover 3 with a sterilized micro plastic feeding port sealing cover 3 with a process sampling tube, putting the process sampling tube into the biofilm culturing chamber 1, extending one end into a set sampling position, connecting the other end with an injector, drawing the injector to absorb a certain amount of water sample and a micro plastic biofilm sample, pulling out the injector after sampling is finished, and replacing the single micro plastic feeding port sealing cover;
and 5, collecting the floating and sinking three-state micro-plastic: the collection and separation process of the micro plastic biofilms comprises the steps that a collecting cylinder baffle plate 9 is pushed and pulled downwards along a side sliding groove of an arc-shaped collecting cylinder 8 to the bottommost end through a lifting rod B801, the lower end of the arc-shaped collecting cylinder 8 is in a closed state, the arc-shaped collecting cylinder 8 and the collecting cylinder baffle plate 9 are cleaned and sterilized in advance when the collection cylinder 8 is used, the collection cylinder 8 and the collecting cylinder baffle plate 9 are placed into a biofilm culture chamber 1 through a micro plastic adding port 2 in advance, when the arc-shaped collecting cylinder 8 sinks to a preset position in the biofilm culture chamber 1, the collection cylinder baffle plate 9 is pushed downwards along the side sliding groove of the arc-shaped collecting cylinder 8 to the position where the collecting cylinder baffle plate 9 just sinks into the water surface through a lifting rod B901 on the collecting cylinder baffle plate 9, an upper water sample and a suspended micro plastic biofilm in the biofilm culture chamber automatically flow into the arc-shaped collecting cylinder 8, when the upper water sample and the floating micro plastic biofilm are completely collected, the collecting cylinder baffle plate 9 is pulled upwards along the side sliding groove of the arc-shaped collecting cylinder 8, the arc-shaped collecting cylinder 8 is closed, and finally the collection cylinder 8 is pulled upwards through the lifting rod A to be taken out of the arc-shaped collecting cylinder 8 from the biofilm culture chamber 1, and the floating water sample is sucked out by an injector, and the floating water sample is sucked out. And placing the sterilized micro plastic feeding port sealing cover 3 with the process sampling tube from the micro plastic feeding port 2, placing the process sampling tube into the biomembrane culture chamber 1, and collecting the suspended layer biomembrane sample. Finally, opening a sealing cover of a bottom micro-plastic discharge port 11 to enable bottom layer liquid to flow out from the lower part, and collecting a biological membrane sample of a sediment layer;
step 6, dismantling and cleaning the device: after the cultivation, close aeration structure and accuse temperature structure, collect the separation with the inside biomembrane sample collection of biomembrane culture room 1 through semi-automatization collection separation structure, cultivate the biomembrane with the room 1 and support base 7 separation, then upwards promote biomembrane culture room 1, biomembrane culture room 1 bottom is equipped with gaseous aeration incoming port 12, and gaseous aeration incoming port 12 cup joints each other with aeration pipe 14, along with the upwards promotion of biomembrane culture room 1, be convenient for to the separation of gaseous aeration incoming port 12 and aeration pipe 14, carry out the separation that biomembrane culture room 1 and little plastics interpolation mouth 3 and bottom little plastics discharge port 11 sealed lid afterwards, culture apparatus separates promptly and accomplishes, be convenient for to the washing of each biomembrane culture subassembly.
Those skilled in the art will appreciate that the invention may be practiced without these specific details. Although illustrative embodiments of the present invention have been described above to facilitate the understanding of the present invention by those skilled in the art, it should be understood that the present invention is not limited to the scope of the embodiments, and various changes may be made apparent to those skilled in the art as long as they are within the spirit and scope of the present invention as defined and defined by the appended claims, and all matters of the invention which utilize the inventive concepts are protected.

Claims (5)

1. A little plastic biofilm module culture apparatus which characterized in that: the device consists of a plurality of culture boxes, wherein each culture box consists of a biological membrane culture structure, a semi-automatic collection and separation structure, an aeration structure and a temperature control structure;
the biological membrane culture structure comprises a biological membrane culture chamber (1), a micro-plastic adding port (2), a micro-plastic adding port sealing cover (3), a nutrient substance putting port (4) and a micro-plastic substrate (5);
the semi-automatic collecting and separating structure comprises a process sampling tube (6), an arc-shaped collecting cylinder (8), a collecting cylinder baffle (9) and a bottom micro-plastic discharge port (11);
the aeration structure comprises a gas aeration inlet (12), an umbrella-shaped culture solution fluidization air nozzle (13) and an aeration pipeline (14);
the temperature control structure comprises a silica gel heating sheet (15), a temperature control sensor (16) and a key type temperature digital display controller (17);
the top of the biological membrane culture chamber (1) is provided with a micro-plastic adding port (2), a micro-plastic adding port sealing cover (3) is correspondingly arranged, the micro-plastic adding port sealing cover (3) is matched with the micro-plastic adding port (2) to complete sealing, and the nutrient substance feeding port (4) is arranged on one side of the top of the biological membrane culture chamber (1) and is sealed through a rubber cap;
the sealing covers (3) of the micro-plastic adding openings are divided into two types, one type is a single sealing type sealing cover, and the other type is a sampling sealing cover for a sampling tube (6) in the process of penetrating the center of the sealing cover;
the integral arc-shaped collecting cylinder (8) is of a semi-closed arc structure, the arc surface of the cylinder is 240/360 degrees, a sliding groove (10) is formed in the vertical edge of the arc surface and used for installing a collecting cylinder baffle (9), the size of the cylinder top of the arc-shaped collecting cylinder (8) can be matched with the micro plastic adding port (2), and a lifting rod A (801) is fixedly arranged on the cylinder top of the arc-shaped collecting cylinder (8) and used for collecting suspended layer micro plastic biofilm samples in a layered mode;
the collecting cylinder baffle (9) is of a rectangular structure, two sides of the collecting cylinder baffle (9) are matched with the sliding grooves (10) and can be inserted into the sliding grooves to slide, a lifting rod B (901) is fixedly arranged at the top of the collecting cylinder baffle (9), the collecting cylinder baffle (9) is adjusted through the lifting rod B (901) to achieve sampling and closing of the arc-shaped collecting cylinder (8), and the collecting cylinder baffle (9) is matched with the arc-shaped collecting cylinder (8) to achieve sealing of the arc-shaped collecting cylinder (8);
the bottom of the biomembrane culture chamber (1) is provided with a bottom micro-plastic discharge port (11) and two gas aeration access ports (12); the bottom micro plastic discharge port (11) is funnel-shaped, continuously shrinks and narrows from the top to the bottom, and the lower end of the bottom micro plastic discharge port is provided with a sealing cover for sealing and is used for collecting a micro plastic biofilm sample on a sediment layer;
the gas aeration inlet (12) is externally connected with an aeration pipeline (14), and an umbrella-shaped culture solution fluidization air nozzle (13) is connected in the gas aeration inlet (12);
the silica gel heating sheet (15) is an upper layer of insulating silica gel cloth and a lower layer of insulating silica gel cloth, a nichrome wire is laid in the middle of the silica gel heating sheet, and a power line and a temperature control sensor (16) are arranged in the silica gel heating sheet.
2. A module culture apparatus for a micro plastic bio film according to claim 1, wherein: the sealing cover (3) of the micro-plastic feeding port and the sealing cover of the bottom micro-plastic discharge port (11) are connected with the biomembrane culture chamber (1) through a buckle structure so as to achieve the closed state of the biomembrane culture chamber.
3. A module culture apparatus of a micro plastic biofilm according to claim 1, wherein: the utility model discloses a little plastic biomembrane module culture apparatus, including support base (7), support base (7) are open trapezium structure, and the upper end is equipped with the draw-in groove, and the draw-in groove inboard is pasted and is had silica gel heating plate (16), is equipped with the pipeline opening on trapezoidal one side inclined plane for pass aeration pipeline (14), arrange biomembrane culture chamber (1) below in.
4. The method for culturing a micro plastic biofilm of a micro plastic biofilm module culture apparatus according to claim 1, wherein:
step 1, connection of a culture device: connecting a biological membrane culture device, connecting a sealing cover (3) of a micro-plastic feeding port and a sealing cover of a bottom micro-plastic discharge port (11) with a biological membrane culture chamber (1) through a buckle structure so as to achieve the closed state of the biological membrane culture chamber, continuously connecting a gas aeration inlet (12) at the lower end of the biological membrane culture chamber (1) with one end of an aeration pipeline (14), connecting the other end of the aeration pipeline (14) to a compressed gas or an air pump through a support base (7), and separately placing a plurality of culture chambers in clamping grooves of the support base (7);
step 2, culturing: when the culture starts, the culture chambers in the biomembrane culture chamber (1) are mutually separated, a micro-plastic adding port (2) is opened to add culture liquid into each culture chamber, the culture liquid is continuously added until the liquid level reaches four fifths of the height of the culture chamber, and a certain amount of micro-plastic particles are added; if a natural water body is used for culturing, the culture liquid needs to be replaced at set time intervals; if the prepared solution is used for culturing, the nutrient substance feeding port (4) can be added with medicine and nutrient salt solution for mixing, and the prepared solution can be used for overall replacement until the biological membrane is cultured to be mature or the experimental requirements are met;
step 3, setting and simulating culture conditions: simulating the condition culture of a natural water body, providing an aeration culture condition, wherein 2 gas aeration access ports (12) at the lower end of a biomembrane culture chamber (1) are communicated with one end of an aeration pipeline (14), the other end of the aeration pipeline (14) is connected to a compressed gas or an air pump through a supporting base (7), different culture chambers are mutually separated, a switch of an aeration component is turned on to realize the dynamic culture of the micro-plastic biomembrane, and the switch of the aeration component is turned off to realize the static culture of the micro-plastic biomembrane; by controlling the type of the gas introduced, oxygen is introduced into an aerobic biomembrane culture experiment, nitrogen is introduced into an anaerobic biomembrane culture experiment, and air is introduced into a facultative biomembrane culture experiment, so that aerobic, anaerobic and facultative co-culture of the micro-plastic biomembrane is realized; providing temperature control culture conditions, connecting a power supply, turning on a temperature control switch on a key-type temperature digital display controller (17), and carrying out temperature regulation on a silica gel heating sheet (15) through temperature feedback of a temperature control sensor (16), or respectively placing different culture chambers in an incubator and setting temperature for culture;
step 4, collecting samples in culture: sampling in the experimental process of the micro plastic biofilm, taking out a single micro plastic feeding port sealing cover (3), replacing the single micro plastic feeding port sealing cover (3) with a sterilized micro plastic feeding port sealing cover (3) with a process sampling tube, putting the process sampling tube into a biofilm culturing chamber (1), extending one end of the process sampling tube into a set sampling position, connecting the other end of the process sampling tube with an injector, drawing the injector to absorb a certain amount of water sample and a micro plastic biofilm sample, pulling out the injector after sampling is finished, and replacing the single micro plastic feeding port sealing cover;
step 5, collecting the suspension and sinking tri-state micro-plastic: the collection and separation process of the micro plastic biomembrane comprises the steps that a collecting cylinder baffle (9) is pushed and pulled downwards to the bottommost end along an arc-shaped collecting cylinder (8) side sliding groove through a lifting rod B (901), the lower end of the arc-shaped collecting cylinder (8) is in a closed state, the collection cylinder baffle (9) is cleaned and sterilized in advance along the arc-shaped collecting cylinder (8) during use, the collection cylinder (8) is placed into a biomembrane culture chamber (1) through a micro plastic adding port (2) in advance, when the arc-shaped collecting cylinder (8) sinks to a preset position in the biomembrane culture chamber (1), the collection cylinder baffle B (901) is pushed downwards along the arc-shaped collecting cylinder (8) side sliding groove to the position that the collecting cylinder baffle (9) just sinks into the water surface, the upper water sample of the biomembrane culture chamber and the suspended micro plastic biomembrane automatically flow into the arc-shaped collecting cylinder (8) through the lifting rod B (901), the upper water sample and the floating state are completely collected by the upper-layer micro plastic biomembrane, the collection cylinder baffle (9) is pulled upwards along the arc-shaped collecting cylinder (8), and the collection chamber (801) is finally pulled to the upper-shaped collecting cylinder to draw the floating chamber, and the floating injector to draw the upper layer of the collecting cylinder (8) to the upper-shaped collecting cylinder (1). And placing the sterilized micro plastic feeding port sealing cover (3) with the process sampling tube from the micro plastic feeding port (2), placing the process sampling tube into the biomembrane culture chamber (1), and collecting the suspended layer biomembrane sample. Finally, opening a sealing cover of a bottom micro-plastic discharge port (11) to enable bottom liquid to flow out from the lower part, so as to realize collection of a sediment layer biofilm sample;
step 6, dismantling and cleaning the device: after the culture is finished, the aeration structure and the temperature control structure are closed, a biomembrane sample inside the biomembrane culture chamber (1) is collected and separated through the semi-automatic collection and separation structure, the biomembrane culture chamber (1) is separated from the support base (7), then the biomembrane culture chamber (1) is lifted upwards, the bottom of the biomembrane culture chamber (1) is provided with a gas aeration access port (12), the gas aeration access port (12) is sleeved with an aeration pipeline (14), the separation of the gas aeration access port (12) and the aeration pipeline (14) is convenient along with the upward lifting of the biomembrane culture chamber (1), then the separation of the biomembrane culture chamber (1) from the sealing covers of the micro-plastic adding port (3) and the bottom micro-plastic discharge port (11) is carried out, the culture device finishes the separation, and is convenient for cleaning each biomembrane culture component.
5. Use of the apparatus of claim 1 for culturing multiple kinds of plastic biofilms floating, suspending, sinking in water.
CN202211177211.3A 2022-09-26 2022-09-26 Microplastic biofilm module culture device and application thereof Pending CN115478008A (en)

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CN202211177211.3A CN115478008A (en) 2022-09-26 2022-09-26 Microplastic biofilm module culture device and application thereof

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Application Number Priority Date Filing Date Title
CN202211177211.3A CN115478008A (en) 2022-09-26 2022-09-26 Microplastic biofilm module culture device and application thereof

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CN115478008A true CN115478008A (en) 2022-12-16

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