Cable bacteria and long-distance electron transmission research device and method thereof

Abstract

The invention discloses cable bacteria and a long-distance electron transmission research device and method thereof, and belongs to the field of cable bacteria research equipment.The cable bacteria research device comprises a groove body, a static box cover is arranged above the groove body in a covering mode, the groove body comprises an inner-layer groove, a middle-layer groove and an outer-layer groove which are sequentially arranged from inside to outside, the inner-layer groove is filled with sediment samples, and the middle-layer groove is filled with sediment samples; the upper portion of the sediment sample is filled with overlying water, nanowires distributed in a crossed mode are arranged in the sediment sample, the ends of the nanowires penetrate out of the inner groove body and then are located outside the inner groove body, and the diameter of the nanowires is smaller than that of cable bacteria; an emptying pipe, a water inlet pipe and a water outlet pipe are arranged in the tank body, the middle-layer tank is filled with a sediment sample and overlying water with the same height as the inner-layer tank, and the outer-layer tank is a heat preservation layer. The method can be used for culturing and cutting the cable bacteria in the sediment sample, and the problems that the research means of the cable bacteria and the long-distance electron transmission process of the cable bacteria are complex, and the repeatability of experimental results is low are solved.

Description

Technical Field

[0001] This invention relates to the field of equipment for studying cable bacteria, and more particularly to a research apparatus and method for studying cable bacteria and their long-distance electron transmission. Background Technology

[0002] Previous studies suggested that due to mass transfer resistance, deep-seated microorganisms in wetland sediments had difficulty simultaneously accessing electron donors and acceptors, resulting in low activity. Recent research has discovered that a type of filamentous fungus with conductive sheaths (i.e., cable bacteria) can extend the range of microbial activity from the nanoscale to the centimeter scale through long-distance electron transport. This discovery has sparked a surge of research into wetland microecology in recent years.

[0003] However, current research methods on *Cyclopyralid* and its long-distance electron transport processes still have limitations, especially the methods for differentiating the experimental and control groups of *Cyclopyralid* are not yet perfect. Current methods for controlling *Cyclopyralid* activity include hydraulic disturbance, laser cutting, and membrane isolation, but these generally suffer from operational difficulties, high costs, and low reproducibility. Specifically, hydraulic disturbance may cause changes in sediment profile characteristics, resulting in low precision in variable control; laser cutting requires a high thickness of sediment samples, which cannot meet the requirements of larger-scale experiments; and membrane isolation requires maintaining the membrane across the cross-section throughout the experimental period, preventing the simultaneous conduct of other experimental tests (such as microelectrodes), thus lacking flexibility. Based on this, this invention proposes a research device and method for *Cyclopyralid* and its long-distance electron transport, ensuring the stability and reproducibility of the relevant experimental research process. Summary of the Invention

[0004] The purpose of this invention is to provide a research apparatus and method for cable bacteria and their long-distance electron transmission, thereby solving the problems mentioned above.

[0005] To solve the above-mentioned technical problems, the present invention adopts the following technical solution:

[0006] This invention discloses a research device for *Cable Bacteria* and its long-distance electron transmission, comprising a tank body covered by a static enclosure. The tank body includes an inner layer tank, a middle layer tank, and an outer layer tank arranged sequentially from the inside out. The inner layer tank contains an inner layer tank, the middle layer tank is located between the inner and middle layer tanks, and the outer layer tank is located between the outer and middle layer tanks. The inner layer tank is filled with a sediment sample, and the sediment sample is covered with overlying water. Cross-distributed nanowires are disposed within the sediment sample, with the ends of the nanowires extending beyond the inner layer tank. The diameter of the nanowires is smaller than that of the cable bacteria outside the inner layer tank. An empty pipe is installed in the inner layer tank at the interface between the sediment sample and the overlying water. An inlet pipe is installed above the empty pipe. The other ends of the empty pipe and the inlet pipe extend out of the outer layer tank and are located outside the outer layer tank. An outlet pipe is also installed inside the inner layer tank. The outlet section of the outlet pipe extends out of the outer layer tank and its height is level with the overlying water surface. The middle layer tank is filled with sediment sample and overlying water at the same height as the inner layer tank. The outer layer tank serves as an insulation layer.

[0007] Furthermore, valve three and valve two are respectively installed on the drain pipe and the water outlet pipe.

[0008] Furthermore, a water pump and a valve are installed on the water inlet pipe.

[0009] Furthermore, the inner groove has several operating holes for the nanowires to pass through, and a sleeve matching the nanowires is fitted inside the operating holes.

[0010] Furthermore, the intersections of the nanowires are fixed by knotting or fusing.

[0011] Furthermore, the insulation layer includes an electric heating device and an internal circulation device that fills the entire outer layer groove with heat.

[0012] Furthermore, the static enclosure includes a cover body, on which symmetrically arranged locking blocks are provided on the inner sidewall of the cover body, and the top of the inner layer groove is integrally formed with an outer edge that matches the locking blocks, and the bottom of the cover body extends into the overlying water to form a water seal; a gas sampling tube is provided on one side of the cover body above the locking blocks; and a fan assembly is provided on the top of the cover body.

[0013] Furthermore, the gas sampling tube is equipped with valve four.

[0014] Furthermore, the fan assembly includes fan blades, a fan motor that drives the fan blades to rotate, and a battery pack that supplies power to the fan motor.

[0015] A method for using a research device for cable bacteria and its long-distance electron transmission.

[0016] Step 1: Preliminary preparation and environment setup; First, according to the experimental requirements, fill the inner layer tank with sediment samples. During the filling process, place the cross-distributed nanowires inside the sediment samples, ensuring that the ends of the nanowires protrude from the inner layer tank and are located outside it; then, inject overlying water on top of the sediment samples; next, to ensure stable pressure differential, fill the middle layer tank with sediment samples and overlying water of the same height; finally, set and maintain the target temperature in the outer layer tank.

[0017] Step 2: Experimental scheme initiation and water sample collection; Select the appropriate scheme to start the experiment according to the experimental requirements, and carry out water sample collection simultaneously: When using the static experimental scheme, inject overlying water through the inlet pipe, periodically collect overlying water through the inlet pipe, and drain the overlying water through the drain pipe after collection; When using the dynamic experimental scheme, inject overlying water through the inlet pipe, control the height of overlying water through the outlet pipe and collect overlying water, and drain the overlying water through the drain pipe after collection.

[0018] Step 3, sediment cutting and disturbance control; During the experiment, one end of the nanowire is pulled at a preset frequency, and the other three nanowires are moved by the tension balance, so that the nanowire perpendicular to the pulled end horizontally cuts the sediment sample at the preset position; Because the diameter of the nanowire is smaller than that of the cable bacterium, this cutting method will not significantly disturb other microorganisms other than the cable bacterium and the physicochemical characteristics of the sediment sample profile, cut off the vertically growing cable bacterium, and suspend the long-distance electron transport of the cable bacterium;

[0019] Step 4, Gas Sample Collection; According to experimental requirements, fasten the static chamber cover to the inner tank and turn on the fan assembly; When collecting gas samples according to the experimental cycle requirements, collect samples through the gas sampling tube for subsequent gas analysis.

[0020] Step 5: Analysis of the physicochemical properties of sediment profiles; depending on the experimental requirements, when analyzing the physicochemical properties of sediment profiles, the static box cover must be removed first, and tests should be conducted during the intermittent periods of nanowire cutting using gradient diffusion thin film technology or microelectrode technology.

[0021] Step 6: Sediment sample collection and analysis; according to experimental requirements, drain the overlying water in the inner tank through the drain pipe and water outlet pipe, and then collect sediment samples for analysis.

[0022] Compared with the prior art, the beneficial technical effects of the present invention are as follows:

[0023] The present invention provides a research apparatus and method for studying cable bacteria and their long-distance electron transport. This apparatus and method can be used to cultivate and cut cable bacteria in sediment samples and can improve the compatibility with related research methods. It solves the problems of complex research methods and low reproducibility of experimental results in the study of cable bacteria and their long-distance electron transport process. Attached Figure Description

[0024] The present invention will be further described below with reference to the accompanying drawings.

[0025] Figure 1 This is a structural diagram of the research device for cable bacteria and its long-distance electron transmission according to the present invention;

[0026] Figure 2 This is a schematic diagram of the tank structure;

[0027] Figure 3 This is a schematic diagram of the static enclosure structure;

[0028] Figure 4 This is a schematic diagram of the cable cutting process.

[0029] Figure 5 This is a diagram of the initial state of the nanowires;

[0030] Figure 6 This is a diagram showing the state at which the nanowires begin to be cut.

[0031] Figure 7 This is a diagram showing the state of the nanowires after being cut into a half-plane.

[0032] Figure 8 This is a diagram showing the reverse cutting state of the nanowires;

[0033] Explanation of reference numerals in the attached diagram: 1. Inner layer groove; 2. Middle layer groove; 3. Outer layer groove; 4. Nanofiber; 5. Static enclosure; 6. Intact cable mold; 7. Cut cable mold;

[0034] 1-1 Sediment sample; 1-2 Overlying water; 1-3 Drain pipe; 1-4 Operating port; 1-5 Casing; 1-6 Inlet pipe; 1-7 Outlet pipe; 1-8 Water pump; 1-9 Valve 1; 1-10 Valve 2; 1-11 Valve 3;

[0035] 3-1. Electric heating device; 3-2. Internal circulation device;

[0036] 4-1. Intersection;

[0037] 5-1. Block; 5-2. Gas sampling tube; 5-3. Valve 4; 5-4. Fan assembly. Detailed Implementation

[0038] like Figure 1As shown, a research device for cable bacteria and its long-distance electronic transmission includes a tank. A static enclosure 5 is provided above the tank. The tank includes an inner tank, a middle tank, and an outer tank installed sequentially from the inside to the outside. The inner tank contains an inner tank 1, the middle tank is located between the inner and middle tanks, and the outer tank is located between the middle and outer tanks. The inner, middle, and outer tanks have the same height, and the volumes of the inner, middle, and outer tanks increase sequentially.

[0039] The inner layer groove 1 is filled with sediment sample 1-1, and the sediment sample 1-1 is filled with top layer water 1-2. Cross-distributed nanowires 4 are installed within the sediment sample 1-1. The intersection points 4-1 of the nanowires 4 are fixed by knotting or fusion, ensuring that pulling one end will move the remaining nanowires 4. The ends of the nanowires 4 extend out of the inner layer groove 1 and are located outside the inner layer groove. The diameter of the nanowires 4 is smaller than that of the cable-cutting bacteria. Several operating holes 1-4 are provided on the inner layer groove for the nanowires 4 to pass through. A sleeve 1-5 matching the nanowire 4 is fitted inside each operating hole 1-4. The sleeve 1-5 is made of silicone or similar material, providing both a sealing effect and ensuring the operation of the nanowires 4. Furthermore, after exiting the inner layer groove 1, the nanowires 4 extend into the middle layer groove 2 and are fixed by springs, rubber bands, etc., ensuring that the portion within the sediment sample 1-1 remains taut, thus guaranteeing the cutting of the cable-cutting bacteria.

[0040] The inner tank 1 is located at the junction of the sediment sample 1-1 and the overlying water 1-2, and is equipped with a drain pipe 1-3 for draining the overlying water 1-2. An inlet pipe 1-6 is installed above the drain pipe 1-3, preferably at a height that does not disturb the sediment sample 1-1 when water is introduced. A water pump 1-8 and a valve 1-9 are installed on the inlet pipe 1-6 for water intake. The other ends of the drain pipe 1-3 and the inlet pipe 1-6 extend out of the outer tank 3 and are located outside the outer tank body. An outlet pipe 1-7 is also installed inside the inner tank 1. The outlet section of the outlet pipe 1-7 extends out of the outer tank body and its height is flush with the liquid level of the overlying water 1-2. The liquid level of the overlying water 1-2 is adjusted through the outlet pipe 1-7. The drain pipe 1-3, outlet pipe 1-7, and inlet pipe 1-6 are all perpendicular to the side wall of the tank body.

[0041] The middle layer trough 2 is filled with sediment sample 1-1 and overlying water 1-2 at the same height as the inner layer trough 1 to ensure stable pressure difference.

[0042] The outer tank 3 is an insulation layer, which includes an electric heating device 3-1 and an internal circulation device 3-2 that fills the entire outer tank 3 with heat. Specifically, the electric heating device 3-1 uses an ARH-J variable frequency heating rod as the temperature control component, and its core technical features are PID variable frequency temperature control (accuracy ±0.1℃), dry-burn power-off protection, and resistance to sulfide corrosion. The internal circulation device 3-2 uses an XQP-300 circulation pump, and its core technical features are variable frequency adjustable flow rate (120-300L / h), operating noise ≤30dB, and low-voltage waterproof power supply (12V). Its inlet and outlet are connected to the insulation layer, and together with the electric heating device 3-1, it provides a stable temperature field of 29±0.1℃ for cable bacteria growth and electron transmission experiments, avoiding temperature fluctuations and ensuring the stability of the experimental environment.

[0043] like Figure 2 As shown, the static enclosure 5 includes a cover body located between the inner layer trough 1 and the intermediate trough 2. Clamping blocks 5-1 are symmetrically installed on the inner sidewall of the cover body. The top of the inner layer trough has an integrally formed outer edge that matches the clamping blocks 5-1, and the bottom of the cover body extends into the upper water layer 1-2 to form a water seal. A gas sampling tube 5-2 is installed on one side of the cover body above the clamping blocks 5-1, and a valve 5-3 is installed on the gas sampling tube 5-2. A fan assembly 5-4 is installed on the top of the cover body. The fan assembly 5-4 includes fan blades, a fan motor that drives the fan blades to rotate, and a battery pack that supplies power to the fan motor. Specifically, the power of the fan assembly 5-4 is determined according to experimental requirements, but its airflow should not cause significant disturbance to the upper water layer 1-2 in the inner layer trough 1.

[0044] The method of using a research device for cable bacteria and its long-distance electron transmission is as follows:

[0045] Step 1: Preliminary preparation and environment setup; First, according to the experimental requirements, fill the inner layer tank 1 with sediment sample 1-1. During the filling process, the cross-distributed nanowires 4 should be placed inside the sediment sample 1-1, and the ends of the nanowires 4 should be inserted through the inner layer tank and fixed outside it. The nanowires 4 should be in a taut state; then, inject the overlying water 1-2 into the upper part of the sediment sample 1-1; then, to ensure stable pressure difference, fill the middle layer tank 2 with sediment sample 1-1 and overlying water 1-2 of the same height; finally, turn on the electric heating device 3-1, set the target temperature in the outer layer tank 3, and turn on the internal circulation device 3-2 to maintain the temperature balance in the insulation layer;

[0046] Step 2: Experimental Procedure Start-up and Water Sample Collection; Select the appropriate procedure to start the experiment according to experimental requirements, and simultaneously carry out water sample collection: When using the static experimental procedure, inject the overlying water 1-2 through the inlet pipe 1-6, and keep valve 1-9 normally closed; when water samples need to be collected, periodically (e.g., after injecting the overlying water 1-2, it needs to stand for 1-2 days) open valve 1-9 and collect the overlying water 1-2 through the inlet pipe 1-6, and close valve 1-9 after the collection is completed; after the periodic collection is completed, open valve 3-11 and drain the overlying water 1-2 through the drain pipe 1-3;

[0047] When using the dynamic experimental scheme, open valve 1-9 and valve 2-10, inject the overlying water 1-2 through the inlet pipe 1-6, and control the height of the overlying water 1-2 through the outlet pipe 1-7; when water samples need to be collected, collect the overlying water 1-2 through the outlet pipe 1-7, and use water pump 1-8 to pump subsequent overlying water 1-2 from the preset water source into the inner tank 1 through the inlet pipe 1-6; after the collection is completed, close valve 1-9 and valve 2-10, open valve 3-11, and drain the overlying water 1-2 through the drain pipe 1-3.

[0048] If a static experimental scheme is adopted, after injecting initial overlying water 1-2 into the inner tank 1, keep valve 1-9 normally closed. When water samples need to be collected, at the beginning of the cycle, open valve 1-9 to introduce water through inlet pipe 1-6, and at the end of the cycle, open valve 1-11 to drain the water through drain pipe 1-13. When collecting water samples, valve 1-9 can be opened, and samples can be collected at any time through inlet pipe 1-6. After collection, valve 1-9 can be closed. If a dynamic experimental scheme is adopted, after injecting initial overlying water 1-2 into the inner tank 1, open valve 1-9, valve 2-10, and valve 3-11. Use water pump 1-8 to pump subsequent overlying water 1-2 into the inner tank 1 from a preset water source through water inlet pipe 1-6. At the same time, keep valve 2-10 open, control the liquid level and discharge water through outlet pipe 1-7. When drainage is required, open valve 3-11 to drain the water through drain pipe 1-13. Water samples can be collected through outlet pipe 1-7.

[0049] Step 3, Sediment Cutting and Disturbance Control; During the experiment, one end of nanowire 4 is pulled at a preset frequency. The tension balance effect drives the other three nanowire segments 4 to move, causing the nanowire 4 perpendicular to the pulled end to horizontally cut the sediment sample 1-1 at a preset position, as shown below. Figure 4-7 The diagram shows the cutting direction and area of ​​nanowire 4 during its movement. Because the diameter of nanowire 4 is smaller than that of *C. cableiformis*, this cutting method does not significantly disturb other microorganisms besides *C. cableiformis* or the physicochemical characteristics of the sediment sample 1-1 profile. It cuts off the vertically growing *C. cableiformis*, suspending its long-distance electron transport (e.g., ...). Figure 3 The complete cable bacteria 6 and the cut cable bacteria 7 are shown).

[0050] Step 4, Gas Sample Collection; According to experimental requirements, fasten the static chamber cover 5 with valve 4 5-3 in the open position on the inner tank. After turning on the fan assembly 5-4, close valve 4 5-3. When collecting gas samples according to the experimental cycle requirements, open valve 4 5-3 and collect samples through the gas sampling tube 5-2 for subsequent gas analysis.

[0051] Step 5, analysis of the physicochemical properties of sediment profiles; according to experimental requirements, when analyzing the physicochemical properties of sediment profiles, the static box cover 5 must be removed first, and tests should be conducted using gradient diffusion thin film technology or microelectrode technology during the gaps between the cutting of nanowires 4.

[0052] Step 6: Sediment sample collection and analysis; According to experimental requirements, drain the overlying water 1-2 in the inner layer tank 1 through the drain pipe 1-3 and the water outlet pipe 1-7, and then collect sediment sample 1-1 for analysis.

[0053] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. A research device for cable bacteria and their long-distance electron transmission, characterized in that: The system includes a tank body, with a static enclosure (5) covering the top of the tank body. The tank body comprises an inner tank body, a middle tank body, and an outer tank body arranged sequentially from the inside to the outside. The inner tank body is an inner tank (1), the middle tank body is located between the inner tank body and the middle tank body, and the outer tank body is located between the middle tank body and the outer tank body. The inner tank body (1) is filled with a sediment sample (1-1), and the sediment sample (1-1) is filled with overlying water (1-2). The sediment sample (1-1) is provided with cross-distributed nanowires (4). The ends of the nanowires (4) protrude from the inner tank body (1) and are located outside the inner tank body. The diameter of the nanowires (4) is smaller than that of the cable bacteria. The inner tank (1) is located at the junction of the sediment sample (1-1) and the overlying water (1-2) and is provided with a drain pipe (1-3). A water inlet pipe (1-6) is provided above the drain pipe (1-3). The other ends of the drain pipe (1-3) and the water inlet pipe (1-6) pass through the outer tank (3) and are located outside the outer tank. The inner tank (1) is also provided with a water outlet pipe (1-7). The water outlet section of the water outlet pipe (1-7) passes through the outer tank and its height is level with the liquid level of the overlying water (1-2). The middle tank (2) is filled with sediment sample (1-1) and overlying water (1-2) at the same height as the inner tank (1). The outer tank (3) is a heat insulation layer.

2. The research apparatus for cable bacteria and its long-distance electron transmission according to claim 1, characterized in that: The drain pipe (1-3) and the water outlet pipe (1-7) are respectively equipped with valve three (1-11) and valve two (1-10).

3. The research apparatus for cable bacteria and its long-distance electron transmission according to claim 1, characterized in that: The water inlet pipe (1-6) is equipped with a water pump (1-8) and a valve (1-9).

4. The research apparatus for cable bacteria and its long-distance electron transmission according to claim 1, characterized in that: The inner layer groove has several operating holes (1-4) for the nanowires (4) to pass through, and a sleeve (1-5) matching the nanowires (4) is fitted inside the operating holes (1-4).

5. The research apparatus for cable bacteria and its long-distance electron transmission according to claim 1, characterized in that: The intersections (4-1) of the nanowires (4) are fixed by knotting or fusion.

6. The research apparatus for cable bacteria and its long-distance electron transmission according to claim 1, characterized in that: The insulation layer includes an electric heating device (3-1) and an internal circulation device (3-2) that fills the entire outer layer groove (3) with heat.

7. The research apparatus for cable bacteria and its long-distance electron transmission according to claim 1, characterized in that: The static enclosure (5) includes an enclosure body, on which symmetrically arranged locking blocks (5-1) are provided on the inner sidewall of the enclosure body. The top of the inner layer groove body is integrally formed with an outer edge that matches the locking blocks (5-1), and the bottom of the enclosure body extends into the overlying water (1-2) to form a water seal. A gas sampling tube (5-2) is provided on one side of the enclosure body above the locking blocks (5-1). A fan assembly (5-4) is provided on the top of the enclosure body.

8. The research apparatus for cable bacteria and its long-distance electron transmission according to claim 7, characterized in that: The gas sampling tube (5-2) is equipped with valve four (5-3).

9. The research apparatus for cable bacteria and its long-distance electronic transmission according to claim 7, characterized in that: The fan assembly (5-4) includes fan blades, a fan motor that drives the fan blades to rotate, and a battery pack that supplies power to the fan motor.

10. A method of using a research apparatus for cable bacteria and their long-distance electron transmission, as described in claim 7, characterized in that: Step 1, Preliminary preparation and environment setup; First, according to the experimental requirements, fill the inner layer tank (1) with sediment sample (1-1). During the filling process, the cross-distributed nanowires (4) should be placed inside the sediment sample (1-1), and the ends of the nanowires (4) should be placed outside the inner layer tank after passing through it; then, inject the overlying water (1-2) into the upper part of the sediment sample (1-1); then, in order to ensure the pressure difference is stable, fill the middle layer tank (2) with sediment sample (1-1) and overlying water (1-2) of the same height; finally, set the target temperature in the outer layer tank (3) and maintain it. Step 2: Experimental Scheme Initiation and Water Sample Collection; Select the appropriate scheme to start the experiment according to the experimental requirements, and carry out water sample collection simultaneously: When using the static experimental scheme, inject the overlying water (1-2) through the inlet pipe (1-6), periodically collect the overlying water (1-2) through the inlet pipe (1-6), and drain the overlying water (1-2) through the drain pipe (1-3) after collection; When using the dynamic experimental scheme, inject the overlying water (1-2) through the inlet pipe (1-6), control the height of the overlying water (1-2) through the outlet pipe (1-7) and collect the overlying water (1-2), and drain the overlying water (1-2) through the drain pipe (1-3) after collection. Step 3, sediment cutting and disturbance control; During the experiment, one end of the nanowire (4) is pulled at a preset frequency, and the other three nanowires (4) are moved by the tension balance effect, so that the nanowire (4) perpendicular to the pulling end horizontally cuts the sediment sample (1-1) at the preset position; This cutting method will not significantly disturb other microorganisms other than the cable bacteria and the physicochemical characteristics of the sediment sample (1-1) profile, cut off the vertically growing cable bacteria, and suspend the long-distance electron transmission of the cable bacteria; Step 4, gas sample collection; according to experimental requirements, fasten the static box cover (5) to the inner tank and turn on the fan assembly (5-4); when collecting gas samples according to the experimental cycle requirements, collect the samples through the gas sampling tube (5-2) for subsequent gas analysis; Step 5, analysis of the physicochemical properties of sediment profiles; according to experimental requirements, when analyzing the physicochemical properties of sediment profiles, the static box cover (5) must be removed first, and the test is carried out by gradient diffusion thin film technology or microelectrode technology during the gap period of nanowire (4) cutting. Step 6, sediment sample collection and analysis; according to experimental requirements, drain the overlying water (1-2) of the inner layer tank (1) through the drain pipe (1-3) and the water outlet pipe (1-7), and then collect sediment samples (1-1) for analysis.

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