Aerobic microorganism activity detection device based on micro-nano bubbles and evaluation method

Abstract

The invention discloses a micro-nano bubble-based aerobic microorganism activity detection device and an evaluation method. The micro-nano bubble-based aerobic microorganism activity detection device comprises a gas cylinder (1), a micro-nano bubble generator (2), a water storage tank (3), an electric atomizer (4), a maintenance bin (5), a reciprocating air pump (6), a gas chromatograph (7), a culture dish (8), a nutrient solution injector (9), a temperature sensor (10), an air pressure sensor (11), a constant-temperature water bath kettle (13) and a waterway valve (14). The micro-nano bubble generator (11) generates micro-nano bubble water rich in oxygen, the micro-nano bubble water is sprayed into the maintenance bin (5) through the electric atomizer (4) to culture aerobic microorganisms, the temperature is regulated and controlled through the constant-temperature water bath kettle, a culture solution is injected through the nutrient solution injector (9), and the metabolic activity of the microorganisms is promoted. After curing, gas in the curing bin (5) is collected through a reciprocating type sucking pump (6), and the oxygen consumption and the carbon dioxide generation amount are detected by utilizing a gas chromatograph (7), so that the microbial activity is evaluated.

Description

Technical Field

[0001] This invention relates to the field of microbial activity detection and evaluation technology, specifically to an aerobic microbial activity detection device and evaluation method based on micro-nano bubbles, which is particularly suitable for the quantitative evaluation of microbial activity in dust suppression and coal spontaneous combustion prevention projects in mining environments. Background Technology

[0002] The application of aerobic microorganisms in the field of mining safety has become an important direction for the construction of green mines. Through metabolic activities, they achieve core functions such as dust solidification (e.g., secreting extracellular polymers to form aggregates), oxygen consumption (reducing oxygen concentration in the coal spontaneous combustion environment), and degradation of active components (disrupting the coal spontaneous combustion chain reaction), directly determining the effectiveness of project implementation. However, the dynamic changes in microbial activity are significantly affected by environmental factors (such as mine temperature and humidity, mine dust composition, and harmful gases), requiring precise detection to guide on-site microbial community control.

[0003] Currently, traditional methods for detecting microbial activity mainly rely on liquid-phase culture or agar plate counting in laboratory environments, which suffer from problems such as low oxygen transfer efficiency, insufficient control of gas interference, and lengthy detection cycles. Conventional aeration methods produce bubbles with slow dissolved oxygen transfer rates, leading to inaccurate assessments of microbial metabolic activity. In open or semi-closed culture systems, external air can easily penetrate, affecting the accurate monitoring of oxygen and carbon dioxide concentrations within the culture chamber, especially with low sensitivity to trace gas changes. Plate counting methods require a 3-7 day culture cycle, and liquid-phase OD value methods are severely affected by mineral dust particles, failing to meet the engineering requirements of "rapid feedback and real-time adjustment" in mine operations.

[0004] The potential of micro / nanobubble technology in the detection of aerobic microbial activity remains largely untapped. Current technologies lack a systematic approach that integrates micro / nanobubble oxygen supply, atomized spraying, closed cultivation, and gas chromatography detection. Furthermore, a multi-parameter, gas-liquid-solid phase linkage detection system has not been established, making it difficult to distinguish between gas concentration changes caused by microbial metabolism and physical adsorption, thus hindering rapid and high-precision quantitative assessment of microbial activity. Therefore, there is an urgent need to develop an activity assessment device and method that can enhance oxygen transfer efficiency, precisely control culture conditions, and achieve non-destructive detection of gas components to support the large-scale application of microbial technology in mining safety. Summary of the Invention

[0005] The purpose of this invention is to address the aforementioned problems in the existing technology by providing an aerobic microbial activity detection device based on micro-nano bubbles, thereby solving the problems of low detection efficiency, difficulty in accurately quantifying microbial metabolic activity, interference of external gas components with test results, poor accuracy, and inability to monitor gas component changes in real time in traditional microbial activity detection technologies.

[0006] Another objective of this invention is to provide a method for detecting and evaluating the activity of aerobic microorganisms based on micro-nano bubbles.

[0007] To achieve the above-mentioned objectives of the present invention, the aerobic microbial activity detection device and evaluation method based on micro-nano bubbles of the present invention are implemented using the following technical solutions.

[0008] This invention relates to an aerobic microbial activity detection device based on micro-nano bubbles, characterized by comprising: a gas cylinder, a micro-nano bubble generator, a water tank, an electric nebulizer, a curing chamber, a reciprocating air pump, a gas chromatograph, a petri dish, a nutrient solution syringe, a temperature sensor, a pressure sensor, a constant temperature water bath, and water valves; the gas cylinder is connected to the air inlet of the micro-nano bubble generator via a gas pipe, the water inlet of the micro-nano bubble generator is connected to a water source, and the water outlet of the micro-nano bubble generator is connected to the water tank; the outlet of the water tank is connected to the electric nebulizer via a water supply pipeline, and the water valves are installed on the water supply pipeline to control the on / off state of the water supply pipeline; the electric nebulizer is equipped with an electric nebulizer water storage container; the curing chamber is sealed. The container is made of transparent glass or corrosion-resistant plastic to facilitate observation of the culture process; the aerobic microbial culture dish for inoculation is placed inside the culture chamber, and the outside of the culture chamber is connected to the nutrient solution syringe; the culture chamber is placed in a constant temperature water bath, and the temperature sensor and the pressure sensor are installed inside the culture chamber. The outlet of the culture chamber is connected to a reciprocating air pump through an air pipe, and the outlet of the reciprocating air pump is connected to a gas chromatograph; the outlet of the electric nebulizer is connected to the inlet of the culture chamber through a pipeline; the connecting pipeline between the reciprocating air pump (6) and the culture chamber (5) adopts a sealed design, and the air extraction volume is adjustable; the airtightness of the culture chamber (5) is monitored in real time by the pressure sensor (11), and the air pressure inside the chamber is maintained within ±5% of the outside atmospheric pressure difference.

[0009] Preferably, the reciprocating air pump is a unidirectional pump.

[0010] Preferably, the water storage tank has a volume of 5L to 10L and is used for temporary storage of micro-nano bubble water; the water storage container of the electric atomizer has a fixed capacity of 100mL to 200mL; and the maintenance chamber has a volume of 1L to 5L.

[0011] The present invention employs the following steps in its method for detecting and evaluating the activity of aerobic microorganisms based on micro / nanobubbles:

[0012] S1. Preparation and storage of micro-nano bubble water: Oxygen gas from the gas cylinder is introduced into the micro-nano bubble generator. The gas and water intake are controlled to generate oxygen-rich micro-nano bubble water, which is then temporarily stored in a water tank. The gas cylinder can be replaced to adjust the gas composition in the micro-nano bubble water, for example, by using pure oxygen (concentration not less than 99%) to enhance the dissolved oxygen content.

[0013] S2. Micro-nano bubble water atomization: Micro-nano bubble water in the water storage tank is transported to the electric atomizer through the water supply pipeline for atomization and sprayed into the maintenance chamber; the on-off of the water supply pipeline is adjusted by the water valve to ensure the stability of the atomization process; the atomized particle size is controlled within the range of 50μm to 200μm to increase the contact area and efficiency between the water mist and microorganisms.

[0014] S3. Aerobic microbial culture: Aerobic microorganisms for suppressing mine dust are inoculated onto the culture dish in the curing chamber. Using a nutrient solution injector connected to the outside of the curing chamber, the same amount of culture solution is injected each time by controlling the scale to provide nutrition for the microorganisms. The culture solution contains carbon source, nitrogen source, inorganic salts and growth factors to maintain the normal metabolism of the microorganisms.

[0015] S4. Real-time monitoring and control of the curing environment: The curing chamber is placed in a constant temperature water bath. The temperature inside the curing chamber is regulated by the constant temperature water bath to simulate the mine environment and study the effect of temperature on microbial activity. At the same time, temperature and pressure sensors are used to monitor the temperature and pressure inside the curing chamber in real time. Environmental parameters are monitored and adjusted in real time to ensure stable culture conditions, maintain the airtightness of the curing chamber (the pressure difference with the outside atmospheric pressure is maintained within ±5%), avoid interference from outside air, and improve the accuracy of gas detection.

[0016] S5. Metabolic gas collection: After the set curing time (e.g., 24 hours), use a reciprocating air pump to extract gas from the curing chamber; the reciprocating air pump is a one-way pump to prevent outside air from entering.

[0017] S6. Gas Component Detection: The extracted gas is passed into a gas chromatograph to analyze the gas components and their content, with a focus on detecting changes in the concentration of oxygen and carbon dioxide; the gas chromatograph provides high-precision data for quantifying microbial metabolic activities.

[0018] S7. Microbial Activity Assessment: Based on changes in gas composition, oxygen consumption and carbon dioxide production are calculated, and the microbial respiratory quotient (RQ) is further calculated: RQ = (carbon dioxide production / oxygen consumption) × 100%. When the RQ value is greater than the baseline RQ value for microorganisms cultured in ordinary water, it indicates that micro / nano bubble water has a significant promoting effect on microbial activity. This assessment method is applicable to mining engineering, such as enhancing the effect of microorganisms in inhibiting dust and spontaneous combustion of coal through activity enhancement.

[0019] Furthermore, the water storage tank has a volume of 5L to 10L and is used for temporary storage of micro-nano bubble water to ensure bubble stability; the water storage container of the electric atomizer has a fixed capacity of 100mL to 200mL to ensure consistent water volume for each atomization; and the maintenance chamber has a volume of 1L to 5L.

[0020] Furthermore, in step S4, the constant temperature water bath is used to regulate the temperature inside the curing chamber, with the temperature range controlled between 15℃ and 75℃ and the temperature control accuracy being ±0.1℃.

[0021] Furthermore, in step S3, the culture medium comprises glucose as a carbon source, ammonium salt as a nitrogen source, phosphate as an inorganic salt, and vitamins as growth factors.

[0022] Furthermore, in step S1, the concentration of oxygen in the gas cylinder is not less than 99%; in step S5, the pumping rate of the reciprocating pump is controlled at 10 mL / min to 100 mL / min, and the pumping time is fixed at 1 minute to ensure the representativeness and repeatability of the gas sample.

[0023] Furthermore, the airtightness of the maintenance chamber is monitored in real time by a pressure sensor, and the air pressure inside the chamber is maintained within ±5% of the outside atmospheric pressure difference to avoid gas leakage and ensure the accuracy of gas detection.

[0024] Furthermore, the culture medium injected by the nutrient solution syringe contains glucose as the carbon source at a concentration of 10 g / L to 20 g / L; ammonium sulfate as the nitrogen source at a concentration of 5 g / L to 10 g / L; potassium dihydrogen phosphate as the inorganic salt at a concentration of 1 g / L to 2 g / L; and B vitamins as the growth factor at a concentration of 0.1 g / L to 0.5 g / L, to ensure the consistency of microbial growth.

[0025] In practical applications, the gas cylinder can be replaced with different gases (such as pure oxygen, air, or oxygen-enriched air) to study the effects of different gas sources on microbial activity. The air and water intake of the micro / nano bubble generator can be precisely controlled to optimize the size and stability of the micro / nano bubbles, thereby enhancing oxygen solubility and transfer efficiency. For example, the air intake can be set to 0.00288 m³ / h. 3 / h~0.0033m 3 / h, influent flow rate is 0.0168m³ 3 / h~0.0192m 3 The system operates at a rate of / h to ensure uniform bubble distribution. The electric nebulizer atomizes the droplets to ensure even coverage of the culture dish, increasing the contact area between water and microorganisms. Data from the temperature and pressure sensors are recorded every 5 minutes and adjusted by regulating the parameters of the constant-temperature water bath to study the impact of external factors on microbial activity and improve the applicability of the assay.

[0026] In the aforementioned evaluation method, data from gas chromatography can also be used to calculate the oxygen consumption rate and carbon dioxide production rate of microorganisms. Based on the metabolic characteristics of aerobic microorganisms (consuming oxygen and producing carbon dioxide), the activity of microorganisms can also be quantitatively evaluated by calculating the O2 consumption rate and CO2 production rate, as shown in the following formula:

[0027] Microbial activity index = (ΔCO2 / Δt) / (ΔO2 / Δt)

[0028] Where ΔCO2 and ΔO2 are the volume concentration changes of CO2 and O2 per unit time, respectively, in percentage (%); Δt is the incubation time, in hours (h).

[0029] Compared with existing technologies, this invention is based on the detection of aerobic microbial activity using micro-nano bubbles.

[0030] The device and evaluation method enhance dissolved oxygen transfer efficiency through micro-nano bubble water, combined with atomization technology and closed-loop curing, to achieve efficient and controllable microbial cultivation and gas detection. This provides a reliable evaluation method for engineering applications such as microbial suppression of dust and coal spontaneous combustion in mines. This invention has the advantages of high detection sensitivity, simple operation, and strong environmental adaptability. Specifically, it has the following beneficial effects:

[0031] (1) By using micro-nano bubble technology to enhance oxygen dissolution and transfer efficiency, the problem of slow oxygen dissolution in traditional aeration methods is solved, significantly improving the metabolic activity of aerobic microorganisms and shortening the required culture time.

[0032] (2) Synergistic effect of atomization + micro-nano bubbles: Atomization technology increases the contact area between water and microorganisms, accelerating metabolic reactions. The atomized micro-nano bubble water forms droplets of 50μm~200μm, which not only retains the high dissolved oxygen characteristics of micro-nano bubbles, but also allows microorganisms in every part of the chamber to come into contact with sufficient oxygen and water through the uniform diffusion of droplets, thus solving the problems of uneven contact of microorganisms and local hypoxia in traditional liquid immersion methods.

[0033] (3) Combining a sealed curing chamber and real-time sensors effectively reduces external interference, reduces gas component detection error, achieves accurate detection of gas components, and significantly improves the accuracy of activity assessment.

[0034] (4) The device features modular design, high degree of automation, simple structure, and convenient operation. Precise control of multiple parameters: temperature, air pressure, dissolved oxygen, and nutrient solution supply are all controlled in a closed-loop manner. For example, when the temperature sensor detects a deviation of the chamber temperature from the set value, the constant temperature water bath automatically adjusts; when the air pressure sensor detects an abnormal pressure difference, the system can automatically trigger gas replenishment or exhaust to ensure the stability of the culture environment, and the coefficient of variation of experimental data can be controlled within 5%. By adjusting the gas source, temperature, and culture solution parameters, it is suitable for simulating different microbial strains and mine environments, providing a basis for the large-scale application of microbial technology in mining safety; the device integrates a gas chromatograph and a vacuum pump to achieve automated and continuous gas detection, reducing manual operation and making it suitable for long-term experiments or on-site deployment.

[0035] (5) The evaluation method is based on quantitative gas data (such as RQ value) to provide objective and repeatable activity indicators, providing a scientific basis for optimizing the effect of microorganisms in inhibiting dust and coal spontaneous combustion. Attached Figure Description

[0036] Figure 1 This is a schematic diagram of the system structure of the aerobic microbial activity detection device based on micro-nano bubbles according to the present invention.

[0037] Figure 2 This is a schematic diagram of the electric atomizer structure used in the aerobic microbial activity detection device based on micro-nano bubbles of the present invention.

[0038] Figure 3 This is a flowchart illustrating the principle process of the aerobic microbial activity detection and evaluation method based on micro-nano bubbles according to the present invention.

[0039] The attached diagram is labeled as follows: 1-Gas cylinder; 2-Micro-nano bubble generator; 3-Water storage tank; 4-Electric nebulizer; 5-Cure chamber; 6-Reciprocating air pump; 7-Gas chromatograph; 8-Cultural dish; 9-Nutrient solution syringe; 10-Temperature sensor; 11-Gas pressure sensor; 12-Electric nebulizer water storage container; 13-Constant temperature water bath; 14-Water valve. Detailed Implementation

[0040] The aerobic microbial activity detection device and evaluation method based on micro / nanobubbles of the present invention will be further described clearly and completely below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0041] Depend on Figure 1 The diagram shown is a schematic of the system structure of the aerobic microbial activity detection device based on micro-nano bubbles according to the present invention, and is combined with... Figure 2As can be seen, the aerobic microbial activity detection device based on micro-nano bubbles of the present invention is composed of a gas cylinder 1, a micro-nano bubble generator 2, a water storage tank 3, an electric nebulizer 4, a curing chamber 5, a reciprocating air pump 6, a gas chromatograph 7, a petri dish 8, a nutrient solution syringe 9, a temperature sensor 10, a pressure sensor 11, a constant temperature water bath 13, and a water valve 14 connected together. The gas cylinder 1 is connected to the air inlet of the micro-nano bubble generator 2 via a gas pipe, the water inlet of the micro-nano bubble generator 2 is connected to a water source, and the water outlet of the micro-nano bubble generator 2 is connected to the water storage tank 3. The outlet of the water storage tank 3 is connected to the electric nebulizer 4 via a water supply pipeline, and the water valve 14 is installed on the water supply pipeline for... The electric nebulizer 4, equipped with an electric nebulizer water storage container 12, controls the opening and closing of the water supply pipeline. The curing chamber 5 is a sealed container made of transparent glass or corrosion-resistant plastic to facilitate observation of the cultivation process. An aerobic microbial culture dish 8 is placed inside the curing chamber 5, and a nutrient solution syringe 9 is connected to the outside of the curing chamber 5. The curing chamber 5 is placed in a constant temperature water bath 13. A temperature sensor 10 and a pressure sensor 11 are installed inside the curing chamber 5. The outlet of the curing chamber 5 is connected to a reciprocating air pump 6 via a gas pipe. The outlet of the reciprocating air pump 6 is connected to a gas chromatograph 7. The reciprocating air pump 6 is a unidirectional pump. The outlet of the electric nebulizer 4 is connected to the inlet of the curing chamber 5 via a pipeline. These components achieve integrated operation through pipelines and electrical connections, ensuring a closed and controllable detection process. The airtightness of the curing chamber 5 is monitored in real time by the air pressure sensor 11, and the air pressure inside the chamber is maintained within ±5% of the external atmospheric pressure difference to avoid gas leakage and ensure the accuracy of gas detection; the connecting pipeline between the reciprocating air pump 6 and the curing chamber 5 adopts a sealed design, and the air extraction volume is adjustable; the airtightness of the curing chamber 5 is monitored in real time by the air pressure sensor 11, and the air pressure inside the chamber is maintained within ±5% of the external atmospheric pressure difference.

[0042] The relevant technical parameters are set as follows: the water storage tank 3 has a volume of 5L to 10L and is used for temporary storage of micro-nano bubble water; the water storage container 12 of the electric atomizer has a fixed capacity of 100mL to 200mL; and the maintenance chamber 5 has a volume of 1L to 5L.

[0043] Depend on Figure 3 The diagram shown is a flowchart illustrating the principle and process flow of the aerobic microbial activity detection and evaluation method based on micro / nanobubbles of this invention, combined with... Figure 1 , Figure 2 As can be seen, in the embodiments, the aerobic microbial activity detection device and evaluation method based on micro-nano bubbles of the present invention are implemented using the following technical solutions:

[0044] First, assemble and initialize the device: Connect gas cylinder 1 to the air inlet of micro / nano bubble generator 2 via a gas tube. Gas cylinder 1 can be replaced; for example, pure oxygen (concentration not less than 99%) or air can be used as the gas source to adjust the gas composition in the micro / nano bubble water. Connect the water inlet of micro / nano bubble generator 2 to a water source, controlling the water inlet flow rate to 0.0168 m³ / h. 3 / h~0.0192m 3 / h, intake volume is 0.00288m³ 3 / h~0.0033m 3 The generator produces oxygen-rich micro / nano bubble water at a rate of [per hour]. The outlet of the micro / nano bubble generator 2 is connected to a water storage tank 3, with a volume of 5L–10L, for temporary storage of the micro / nano bubble water to ensure bubble stability. The outlet of the water storage tank 3 is connected to an electric atomizer 4 via a water supply pipeline. A water valve 14 is installed on the water supply pipeline to control its opening and closing. The electric atomizer 4 is equipped with an electric atomizer water storage container 12, with a fixed capacity of 100mL–200mL to ensure consistent water volume per atomization. The atomized particle size is controlled within the range of 50μm–200μm to increase the contact area and efficiency between the water mist and microorganisms.

[0045] The curing chamber 5 is a sealed container with a volume of 1L to 5L, made of transparent glass or corrosion-resistant plastic for easy observation of the cultivation process. Inside the curing chamber 5, a petri dish 8 is placed for inoculating aerobic microorganisms (such as strains used for suppressing mine dust). The curing chamber 5 is externally connected to a nutrient solution injector 9, which is a graduated syringe to ensure that the volume of culture medium injected each time is the same. The culture medium consists of a carbon source (such as glucose, concentration 10g / L to 20g / L), a nitrogen source (such as ammonium sulfate, concentration 1g / L to 2g / L), inorganic salts (such as potassium dihydrogen phosphate, concentration 1g / L to 2g / L), and growth factors (such as B vitamins, concentration 0.1g / L to 0.5g / L) to maintain normal microbial metabolism. The curing chamber 5 is placed in a constant temperature water bath 13. The temperature control accuracy of the constant temperature water bath 13 is ±0.1℃, and the temperature range can be adjusted from 15℃ to 75℃ to simulate the mine environment and study the effect of temperature on microbial activity. Temperature sensor 10 and air pressure sensor 11 are installed inside the curing chamber (5) to monitor the temperature and air pressure inside the chamber in real time. The data is recorded every 5 minutes and adjusted by adjusting the parameters of the constant temperature water bath 13 to ensure that the air pressure inside the chamber is maintained within ±5% of the outside atmospheric pressure difference, so as to avoid gas leakage or outside air intrusion.

[0046] The outlet of the curing chamber 5 is connected to a reciprocating air pump 6 via a gas tube. The reciprocating air pump 6 is a one-way pump to prevent external air from entering. The pumping rate is adjustable within the range of 10 mL / min to 100 mL / min, and the pumping time is fixed at 1 minute to ensure the representativeness and consistency of the gas samples. The outlet of the reciprocating air pump 6 is connected to a gas chromatograph 7 for analyzing gas components and content, with a focus on detecting changes in oxygen and carbon dioxide concentrations.

[0047] The operating procedure is as follows: First, turn on gas cylinder 1 and micro / nano bubble generator 2 to generate micro / nano bubble water and store it in water tank 3; then, open water valve 14, start electric atomizer 4, and spray the micro / nano bubble water into curing chamber 5, with droplets evenly covering petri dishes 8. Inoculate aerobic microorganisms onto petri dishes 8 in curing chamber 5, and inject culture medium through nutrient solution injector 9. The culture medium injected by nutrient solution injector 9 contains glucose as the carbon source (concentration 10g / L~20g / L), ammonium sulfate as the nitrogen source (concentration 5g / L~10g / L), potassium dihydrogen phosphate as the inorganic salt (concentration 1g / L~2g / L), and B vitamins as the growth factor (concentration 0.1g / L~0.5g / L) to ensure consistent microbial growth. After sealing curing chamber 5, start constant temperature water bath 13 and set the required temperature (e.g., 25℃), monitoring environmental parameters in real time through temperature sensor 10 and pressure sensor 11. After a certain period of curing (e.g., 24 hours), the reciprocating air pump 6 is started to extract gas samples from the curing chamber 5 and pass them into the gas chromatograph 7 for analysis.

[0048] Gas detection data was used to assess microbial activity: oxygen consumption and carbon dioxide production were calculated, and the microbial respiratory quotient (RQ) was further calculated, i.e., RQ = (carbon dioxide production / oxygen consumption) × 100%. When the RQ value is greater than the baseline RQ value for microorganisms cultured in ordinary water, it indicates that micro / nano bubble water has a significant promoting effect on microbial activity. In addition, a microbial activity index can be calculated using the formula: Microbial activity index = (ΔCO2 / Δt) / (ΔO2 / Δt), where ΔCO2 and ΔO2 are the volume concentration changes of CO2 and O2 per unit time, respectively, %; and Δt is the culture time, h.

[0049] In this embodiment, oxygen dissolution efficiency is enhanced by micro-nano bubble technology. Combined with atomized spraying and closed curing, rapid and high-precision detection of aerobic microbial activity is achieved. The modular design of the device facilitates parameter adjustment and is suitable for simulating different mine environments, providing a reliable basis for engineering applications of microbial inhibition of dust and coal spontaneous combustion.

[0050] This invention solves the problems of low oxygen supply efficiency, large interference from gas components, and inaccurate results in traditional microbial activity detection. It enhances dissolved oxygen transfer efficiency through micro-nano bubble technology and expands the contact area through atomized spraying, achieving rapid and high-precision activity characterization. This provides a reliable evaluation method for engineering applications such as microbial suppression of dust and coal spontaneous combustion in mines. This invention has the advantages of high detection sensitivity, simple operation, and strong environmental adaptability.

[0051] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Furthermore, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

Claims

1. A device for detecting the activity of aerobic microorganisms based on micro / nanobubbles, characterized in that: It includes a gas cylinder (1), a micro-nano bubble generator (2), a water tank (3), an electric atomizer (4), a curing chamber (5), a reciprocating air pump (6), a gas chromatograph (7), a petri dish (8), a nutrient solution syringe (9), a temperature sensor (10), a pressure sensor (11), a constant temperature water bath (13), and a water valve (14); the gas cylinder (1) is connected to the air inlet of the micro-nano bubble generator (2) through a gas pipe, the water inlet of the micro-nano bubble generator (2) is connected to a water source, and the water outlet of the micro-nano bubble generator (2) is connected to the water tank (3); the outlet of the water tank (3) is connected to the electric atomizer (4) through a water supply pipeline, and the water valve (14) is installed on the water supply pipeline to control the opening and closing of the water supply pipeline; the electric atomizer (4) is equipped with an electric atomizer water storage container (12); the curing chamber (5) is a sealed The container is made of transparent glass or corrosion-resistant plastic to facilitate observation of the culture process; the culture dish (8) for inoculating aerobic microorganisms is placed inside the culture chamber (5), and the outside of the culture chamber (5) is connected to the nutrient solution syringe (9); the culture chamber (5) is placed in a constant temperature water bath (13), and the temperature sensor (10) and the pressure sensor (11) are installed inside the culture chamber (5). The outlet of the culture chamber (5) is connected to a reciprocating air pump (6) through an air pipe, and the outlet of the reciprocating air pump (6) is connected to a gas chromatograph (7); the outlet of the electric nebulizer (4) is connected to the inlet of the culture chamber (5) through a pipeline; the connecting pipeline between the reciprocating air pump (6) and the culture chamber (5) adopts a sealed design, and the air pumping volume is adjustable; the airtightness of the culture chamber (5) is monitored in real time by the pressure sensor (11), and the air pressure inside the chamber is maintained within ±5% of the outside atmospheric pressure difference.

2. The aerobic microbial activity detection device based on micro / nanobubbles as described in claim 1, characterized in that: The reciprocating air pump (6) is a unidirectional pump.

3. The aerobic microbial activity detection device based on micro / nanobubbles as described in claim 1, characterized in that: The water storage tank (3) has a volume of 5L to 10L and is used for temporary storage of micro-nano bubble water; the water storage container (12) of the electric atomizer has a fixed capacity of 100mL to 200mL; and the maintenance chamber (5) has a volume of 1L to 5L.

4. A method for detecting and evaluating the activity of aerobic microorganisms based on micro / nanobubbles, characterized in that: A self-designed aerobic microbial activity detection device based on micro-nano bubbles is used. This device includes a gas cylinder (1), a micro-nano bubble generator (2), a water tank (3), an electric atomizer (4), a curing chamber (5), a reciprocating air pump (6), a gas chromatograph (7), a petri dish (8), a nutrient solution syringe (9), a temperature sensor (10), a pressure sensor (11), a constant temperature water bath (13), and a water valve (14). The gas cylinder (1) is connected to the air inlet of the micro-nano bubble generator (2) via a gas pipe. The water inlet of the micro-nano bubble generator (2) is connected to a water source, and the water outlet of the micro-nano bubble generator (2) is connected to the water tank (3). The outlet of the water tank (3) is connected to the electric atomizer (4) via a water supply pipeline. The water valve (14) is installed... The electric atomizer (4) is equipped with an electric atomizer water storage container (12) installed on the water supply pipeline to control the opening and closing of the water supply pipeline; the maintenance chamber (5) is a sealed container made of transparent glass or corrosion-resistant plastic to facilitate observation of the culture process; the culture dish (8) for inoculating aerobic microorganisms is placed inside the maintenance chamber (5), and the outside of the maintenance chamber (5) is connected to the nutrient solution syringe (9); the maintenance chamber (5) is placed in a constant temperature water bath (13), and the temperature sensor (10) and the pressure sensor (11) are installed inside the maintenance chamber (5). The outlet of the maintenance chamber (5) is connected to the reciprocating air pump (6) through the air pipe, and the outlet of the reciprocating air pump (6) is connected to the gas chromatograph (7). The reciprocating air pump (6) is a one-way pump; the outlet of the electric atomizer (4) is connected to the inlet of the maintenance chamber (5) through the pipeline. The aforementioned method for detecting and evaluating the activity of aerobic microorganisms comprises the following steps: S1. Preparation and storage of micro-nano bubble water: The oxygen in the gas cylinder (1) is introduced into the micro-nano bubble generator (2), the gas intake and water intake are controlled to generate oxygen-rich micro-nano bubble water, and the water flows into the storage tank (3) for temporary storage. S2, Micro-nano bubble water atomization: The micro-nano bubble water in the water storage tank (3) is transported to the electric atomizer (4) through the water supply pipeline for atomization and sprayed into the maintenance chamber (5); the opening and closing of the water supply pipeline is adjusted by the water valve (14) to ensure the stability of the atomization process; the atomization particle size is controlled within the range of 50μm to 200μm; S3. Aerobic microbial culture: Aerobic microorganisms for suppressing mine dust are inoculated on the culture dish (8) in the maintenance chamber (5), and the same amount of culture solution is injected each time by the nutrient solution syringe (9) connected to the outside of the maintenance chamber (5) through the scale control to provide nutrition for the microorganisms; The culture solution contains carbon source, nitrogen source, inorganic salt and growth factor to maintain the normal metabolism of microorganisms. S4. Real-time monitoring and control of the curing environment: The curing chamber (5) is placed in a constant temperature water bath (13). The temperature inside the curing chamber (5) is adjusted by the constant temperature water bath (13) to simulate the mine environment and study the effect of temperature on microbial activity. At the same time, the temperature and air pressure inside the curing chamber (5) are monitored in real time by temperature sensor (10) and air pressure sensor (11). The environmental parameters are monitored and adjusted in real time to ensure stable culture conditions. S5. Metabolic gas collection: After the set curing time, use a reciprocating air pump (6) to extract gas from the curing chamber (5); S6. Gas component detection: The extracted gas is passed into the gas chromatograph (7) to analyze the gas components and content, with a focus on detecting changes in the concentration of oxygen and carbon dioxide; the gas chromatograph (7) provides high-precision data for quantifying microbial metabolic activities; S7. Microbial activity assessment: ① Based on changes in gas composition, calculate oxygen consumption and carbon dioxide production, and further calculate the microbial respiratory quotient (RQ): RQ = (carbon dioxide production / oxygen consumption) × 100%). ② Based on the metabolic characteristics of aerobic microorganisms, the activity of microorganisms is quantitatively assessed by calculating the O2 consumption rate and CO2 generation rate, as shown in the following formula: Microbial activity index = (ΔCO2 / Δt) / (ΔO2 / Δt) Where ΔCO2 and ΔO2 are the volume concentration changes of CO2 and O2 per unit time, respectively, in percentage (%); Δt is the incubation time, in hours (h).

5. The method for detecting and evaluating the activity of aerobic microorganisms based on micro / nanobubbles as described in claim 4, characterized in that: The water storage tank (3) has a volume of 5L to 10L and is used for temporary storage of micro-nano bubble water; the water storage container (12) of the electric atomizer has a fixed capacity of 100mL to 200mL; and the maintenance chamber (5) has a volume of 1L to 5L.

6. The method for detecting and evaluating the activity of aerobic microorganisms based on micro / nanobubbles as described in claim 4, characterized in that: In step S4, the constant temperature water bath (13) is used to regulate the temperature inside the curing chamber (5), with the temperature range controlled between 15℃ and 75℃ and the temperature control accuracy being ±0.1℃.

7. The method for detecting and evaluating the activity of aerobic microorganisms based on micro / nanobubbles as described in claim 4, characterized in that: In step S3, the culture medium consists of glucose as a carbon source, ammonium salt as a nitrogen source, phosphate as an inorganic salt, and vitamins as growth factors.

8. The method for detecting and evaluating the activity of aerobic microorganisms based on micro / nanobubbles as described in claim 4, characterized in that: In step S1, the concentration of oxygen in the gas cylinder (1) is not less than 99%; in step S5, the pumping rate of the reciprocating pump (6) is controlled at 10 mL / min to 100 mL / min, and the pumping time is fixed at 1 minute to ensure the representativeness and repeatability of the gas sample.

9. The method for detecting and evaluating the activity of aerobic microorganisms based on micro / nanobubbles as described in claim 4, characterized in that: The airtightness of the maintenance chamber (5) is monitored in real time by a pressure sensor (11). The pressure inside the chamber is maintained within ±5% of the external atmospheric pressure difference to avoid gas leakage and ensure the accuracy of gas detection.

10. The method for detecting and evaluating the activity of aerobic microorganisms based on micro / nanobubbles as described in claim 4, characterized in that: The culture medium injected by the nutrient solution syringe (9) contains glucose as the carbon source at a concentration of 10 g / L to 20 g / L; ammonium sulfate as the nitrogen source at a concentration of 5 g / L to 10 g / L; potassium dihydrogen phosphate as the inorganic salt at a concentration of 1 g / L to 2 g / L; and B vitamins as the growth factor at a concentration of 0.1 g / L to 0.5 g / L, to ensure consistent microbial growth.

Images