A multi-sequence fidelity monitoring device and method for deep-sea microbial culture

Abstract

The invention discloses a multi-sequence fidelity monitoring device and method for deep-sea microorganism cultivation, which relates to the technical field of marine microorganisms, and comprises a gas boosting unit, a pressure-maintaining cultivation unit, a fidelity monitoring unit and a collection control unit; the gas boosting unit is used to provide high-pressure nitrogen to the pressure-maintaining cultivation unit and the fidelity monitoring unit, so as to realize a high-pressure cultivation environment for deep-sea cultivation samples, and to form a pressure difference by changing the pressure value of nitrogen injected into a pressure-maintaining cultivation cylinder and a pressure-maintaining buffer cylinder, so as to push a piston to realize the pressure-maintaining transfer of deep-sea cultivation samples; the switching sampling work of the multi-sequence pressure-maintaining cultivation cylinder is realized by opening and closing a second multi-way valve, and the demand for in-situ pressure-maintaining sampling and monitoring of multiple pressure-maintaining cultivation cylinders is met by using a single set of fidelity monitoring units, thereby reducing the equipment cost; the whole process of pressure-maintaining operation of sampling and recovering cultivation samples is realized by opening and closing a three-way valve and a four-way valve, thereby ensuring the authenticity of the cultivation samples in the monitoring process, reducing the loss of the cultivation samples in the monitoring process, improving the sampling frequency and efficiency of the cultivation process, and extending the cultivation cycle of deep-sea microorganisms.

Description

Technical Field

[0001] The present invention relates to the technical field of marine microorganisms, and more specifically, to a multi-sequence fidelity monitoring device and method for deep-sea microorganism cultivation. Background Art

[0002] The deep sea contains rich microbial resources, and its unique biochemical processes and metabolites are key links in exploring the biogeochemical cycles of extreme environments and even the origin of life. High pressure is an important environmental indicator that distinguishes the deep sea from other ecosystems. This characteristic makes deep-sea microorganisms generally pressure-resistant. Therefore, deep-sea microorganisms are closely related to pressure. When leaving the high-pressure environment, the growth and metabolic capacity of microorganisms may be seriously affected, and they may enter a dormant state or even rupture and die, which greatly limits the research and application of deep-sea microorganisms. Existing deep-sea microbial culture research mainly maintains the growth and metabolic activity of microorganisms through pressure-maintaining culture. However, during the sample monitoring process, the microbial culture samples must still be sampled after decompression for monitoring. Due to the sudden change in pressure during decompression, the microbial culture samples taken out of the normal pressure conditions for monitoring will cause microbial cell rupture and other phenomena, resulting in sample distortion. At the same time, since the key stages of the microbial culture process require high-frequency monitoring, the loss of the total amount and nutrients of the culture system caused by continuous decompression sampling cannot be ignored for the culture system. Summary of the invention

[0003] In order to overcome the defects of the above-mentioned prior art in deep-sea microbial culture monitoring operations, such as sample distortion and large sampling loss caused by the need for pressure relief sampling, the present invention provides a multi-sequence fidelity monitoring device and method for deep-sea microbial culture. The present invention performs low-loss sample transfer and fidelity monitoring on the multi-sequence culture system under pressure maintenance throughout the whole process, thereby meeting the high-fidelity, low-loss and multi-parameter monitoring of the culture samples and extending the pressure maintenance culture periodicity of the samples.

[0004] In order to solve the above technical problems, the technical solution of the present invention is as follows:

[0005] The present invention provides a multi-sequence fidelity monitoring device for deep-sea microbial culture, comprising a gas pressurization unit, a pressure-maintaining culture unit, a fidelity monitoring unit and a collection control unit;

[0006] The pressure-maintaining culture unit comprises a first multi-way valve, a plurality of pressure-maintaining culture cylinders and a second multi-way valve; the input end of the first multi-way valve is connected to the first output end of the gas pressurization unit, the plurality of output ends of the first multi-way valve are respectively connected to one end of a pressure-maintaining culture cylinder, and the other end of each pressure-maintaining culture cylinder is respectively connected to an input end of the second multi-way valve;

[0007] The fidelity monitoring unit includes a three-way valve, a four-way valve, a pressure-maintaining buffer cylinder and a monitoring compartment; the first port of the four-way valve is connected to the output end of the second multi-way valve, and the second port of the four-way valve is connected to the input end of the monitoring compartment; the third port of the four-way valve is connected to one end of the pressure-maintaining buffer cylinder, and the other end of the pressure-maintaining buffer cylinder is connected to the first port of the three-way valve; the fourth port of the four-way valve is connected to the second port of the three-way valve, and the third port of the three-way valve is connected to the second output end of the gas pressurizing unit;

[0008] The input end of the acquisition control unit is connected to the data output end of the monitoring compartment, and the output end of the acquisition control unit is respectively connected to the control end of the gas booster unit, the control end of the first multi-way valve, the control end of the second multi-way valve, the control end of the three-way valve, and the control end of the four-way valve.

[0009] The gas pressurization unit is used to provide high-pressure nitrogen to the pressure-maintaining culture unit and the fidelity monitoring unit to achieve a high-pressure culture environment for deep-sea culture samples; the gas pressurization unit realizes the transfer of deep-sea culture samples between the pressure-maintaining culture cylinder, the pressure-maintaining buffer cylinder and the monitoring compartment by changing the nitrogen pressure value injected into the pressure-maintaining culture cylinder and the pressure-maintaining buffer cylinder; the switching sampling of multi-sequence pressure-maintaining culture cylinders is realized by opening and closing the second multi-way valve, and the sampling and monitoring requirements of multiple pressure-maintaining culture cylinders are met by using a single set of fidelity monitoring units, thereby reducing equipment costs; the whole process of pressure-maintaining operation of sampling and recovery of culture samples is realized by opening and closing the three-way valve and the four-way valve, thereby ensuring the authenticity of the culture samples during the monitoring process, reducing the loss of culture samples during the monitoring process, improving the sampling frequency and efficiency of the culture process, and extending the culture cycle of deep-sea microorganisms.

[0010] Preferably, each of the pressure-maintaining culture cylinders comprises a gas-sealed end cap, a first cavity, a first piston and a seawater-sealed end cap;

[0011] The first piston is arranged inside the first cavity, dividing the first cavity into a first gas pressurization cavity and a pressure-maintaining culture cavity; the gas sealing end cover is arranged at one end of the first gas pressurization cavity, and the seawater sealing end cover is arranged at one end of the pressure-maintaining culture cavity;

[0012] The output ends of the first multi-way valve are respectively connected to the gas sealing end cover of a pressure-maintaining culture cylinder, and the seawater sealing end cover of each pressure-maintaining culture cylinder is respectively connected to an input end of the second multi-way valve.

[0013] The first cavity is a titanium alloy cylindrical structure with a movable first piston built in. The first piston is covered with an O-ring to isolate the first cavity into a first gas pressurization cavity and a pressure-maintaining culture cavity. The gas pressurization unit injects high-pressure nitrogen into the first gas pressurization cavity of the pressure-maintaining culture cylinder through a first multi-way valve to achieve a high-pressure culture environment for deep-sea culture samples.

[0014] Preferably, the pressure-maintaining buffer cylinder comprises a first sealing end cover, a second cavity, a second piston and a second sealing end cover;

[0015] The second piston is arranged inside the second cavity, dividing the second cavity into a second gas pressurization cavity and a pressure-maintaining buffer cavity; the first sealing end cover is arranged at one end of the pressure-maintaining buffer cavity, and the second sealing end cover is arranged at one end of the second gas pressurization cavity;

[0016] The third port of the four-way valve is connected to the first sealing end cover of the pressure-maintaining buffer cylinder, and the second sealing end cover of the pressure-maintaining buffer cylinder is connected to the first port of the three-way valve.

[0017] The second cavity is a titanium alloy cylindrical structure with a built-in movable second piston. The second piston is sleeved with an O-ring, which divides the second cavity into a second gas boosting cavity and a pressure-maintaining buffer cavity. The gas boosting unit injects high-pressure nitrogen into the second gas boosting cavity of the pressure-maintaining buffer cylinder through a three-way valve to achieve a high-pressure culture environment for deep-sea culture samples. The culture samples in the pressure-maintaining culture cylinder enter the pressure-maintaining buffer cavity of the pressure-maintaining culture cylinder through the second multi-way valve and the four-way valve to achieve pressure-maintaining caching of the culture samples.

[0018] Preferably, the monitoring compartment comprises a third sealing end cover, a fourth sealing end cover, a compartment body, a window, a Raman spectroscopic probe, an ultraviolet spectroscopic probe, a compartment pressure sensor, a water quality sensor and a liquid level sensor;

[0019] The third sealing end cover and the fourth sealing end cover are respectively arranged at the two ends of the cabin body, and the viewing window is arranged on the side wall of the cabin body; the liquid level sensors are arranged on the lower surface of the third sealing end cover, the water quality sensor and the compartment pressure sensor are arranged on the upper surface of the fourth sealing end cover, and the Raman spectrometer probe and the ultraviolet spectrometer probe are arranged on the viewing window;

[0020] The second port of the four-way valve is connected to the lower surface of the fourth sealing end cover of the monitoring compartment;

[0021] The data output ends of the Raman spectrum probe, the ultraviolet spectrum probe, the water quality sensor and the liquid level sensor are all connected to the input end of the acquisition control unit.

[0022] The cabin is a titanium alloy cylindrical structure. The fourth sealing end cover is connected to the four-way valve for the injection and outflow of culture samples. The compartment pressure sensor and liquid level sensor are used to monitor the pressure value and liquid level of the culture samples in the cabin in real time, and are used to reflect the sample quantity in the cabin. The Raman spectrometer probe, ultraviolet spectrometer probe and water quality sensor monitor the various physical and chemical parameters of the culture samples in the cabin in real time. The gas booster unit injects high-pressure nitrogen into the cabin through the three-way valve and the four-way valve to achieve a high-pressure monitoring environment for deep-sea culture samples.

[0023] Preferably, the water quality sensor includes any one or more of a methane sensor, a carbon dioxide sensor, a dissolved oxygen sensor, a temperature sensor, a conductivity sensor, and a pH sensor.

[0024] The methane sensor, carbon dioxide sensor, dissolved oxygen sensor, temperature sensor, conductivity sensor and pH sensor are used to detect the methane concentration, carbon dioxide concentration, dissolved oxygen concentration, temperature, conductivity and pH value of the culture sample, respectively.

[0025] Preferably, the dissolved oxygen sensor, temperature sensor, conductivity sensor and pH sensor are all probe sensors.

[0026] The probe-type sensor can reduce the area occupied on the fourth sealing end cover, making it easier to lay out.

[0027] Preferably, the device further comprises a plurality of sampling needle valves and a plurality of venting needle valves;

[0028] A vent needle valve is arranged between each output end of the first multi-way valve and one end of a pressure-maintaining culture cylinder;

[0029] A vent needle valve is provided between the other end of the pressure-maintaining buffer cylinder and the first port of the three-way valve;

[0030] A sampling needle valve is arranged between the other end of each pressure-maintaining culture cylinder and an input end of the second multi-way valve;

[0031] A sampling needle valve is provided between the third port of the four-way valve and one end of the pressure-maintaining buffer cylinder;

[0032] A sampling needle valve is arranged between the second port of the four-way valve and the input end of the monitoring compartment.

[0033] Preferably, the device further comprises a plurality of pressure sensors;

[0034] Each of the pressure sensors is correspondingly arranged at a sampling end of a sampling needle valve, and a data output end of each of the pressure sensors is connected to an input end of the acquisition control unit.

[0035] The ventilation needle valve is used to realize the pressurization operation of the pressure-maintaining culture cylinder and the pressure-maintaining buffer cylinder, and the sampling needle valve is used to realize the pressure-maintaining sampling operation of the pressure-maintaining culture cylinder, the pressure-maintaining buffer cylinder and the monitoring subcompartment; the pressure sensor is used to indicate the real-time pressure of the pressure-maintaining culture cylinder, the pressure-maintaining buffer cylinder and the monitoring subcompartment.

[0036] The present invention also provides a multi-sequence fidelity monitoring method for deep-sea microbial culture, which is applied to the above-mentioned device and comprises:

[0037] S1: The gas pressurization unit is connected to all the pressure-maintaining culture cylinders through the first multi-way valve, connected to the pressure-maintaining buffer cylinder through the three-way valve, connected to the monitoring compartment through the three-way valve and the four-way valve, and all the pressure-maintaining culture cylinders, pressure-maintaining buffer cylinders and monitoring compartments are maintained at the first pressure value through nitrogen pressurization;

[0038] S2: Selecting a pressure-maintaining culture cylinder to be monitored, opening the output end corresponding to the first multi-way valve, and causing the gas pressurizing unit to pressurize the pressure-maintaining culture cylinder to be monitored with nitrogen, so that the pressure of the first gas pressurizing chamber is increased to a second pressure value;

[0039] S3: Open the input end and the output end corresponding to the second multi-way valve, and the first port and the third port of the four-way valve, to connect the pressure-maintaining culture cylinder and the pressure-maintaining buffer cylinder; under the pressure difference between the second pressure value and the first pressure value, the first piston in the pressure-maintaining culture cylinder pushes the culture sample in the pressure-maintaining culture chamber to be injected into the pressure-maintaining buffer chamber of the pressure-maintaining buffer cylinder;

[0040] S4: closing the input end and the output end corresponding to the second multi-way valve and the first port of the four-way valve, opening the first port and the third port of the three-way valve, and the gas boosting unit boosts nitrogen to the pressure-maintaining buffer cylinder to increase the pressure of the second gas boosting chamber to a second pressure value;

[0041] S5: Open the second port of the four-way valve to connect the pressure-maintaining buffer cylinder and the monitoring sub-cabin; under the pressure difference between the second pressure value and the first pressure value, the second piston in the pressure-maintaining buffer cylinder injects the culture sample in the pressure-maintaining buffer cavity into the body of the monitoring sub-cabin;

[0042] S6: When the culture sample reaches the liquid level sensor in the monitoring compartment, the second port and the third port of the four-way valve are closed; the Raman spectroscopic probe, the ultraviolet spectroscopic probe, the compartment pressure sensor, and the water quality sensor monitor the culture sample, and transmit the obtained monitoring results to the acquisition control unit;

[0043] S7: The gas pressurizing unit decompresses the pressure-maintaining buffer cylinder to reduce the pressure of the second gas pressurizing chamber to a third pressure value;

[0044] S8: Open the second port and the third port of the four-way valve to connect the pressure-maintaining buffer cylinder and the monitoring compartment; under the pressure difference between the first pressure value and the third pressure value, the second piston in the pressure-maintaining buffer cylinder sucks the culture sample in the monitoring compartment back to the pressure-maintaining buffer chamber, and close the second port and the third port of the four-way valve;

[0045] S9: The gas pressurizing unit pressurizes the pressure-maintaining buffer cylinder with nitrogen, so that the pressure of the second gas pressurizing chamber is increased to the second pressure value; the gas pressurizing unit depressurizes the pressure-maintaining culture cylinder to be monitored, so that the pressure of the first gas pressurizing chamber is reduced to the first pressure value;

[0046] S10: opening the input end and the output end corresponding to the second multi-way valve, and the first port and the third port of the four-way valve, connecting the pressure-maintaining culture cylinder and the pressure-maintaining buffer cylinder; under the pressure difference between the second pressure value and the first pressure value, the second piston in the pressure-maintaining buffer cylinder pushes the culture sample in the pressure-maintaining buffer cavity to be injected into the pressure-maintaining culture cavity of the pressure-maintaining buffer cavity;

[0047] S11: closing the input end and the output end corresponding to the second multi-way valve (4), and the first port and the third port of the four-way valve (5), isolating the pressure-maintaining culture cylinder (3), and completing single-sequence fidelity monitoring;

[0048] S12: Update the selected pressure-maintaining culture cylinder (3) to be monitored, and repeat steps S2-S11 to achieve multi-sequence fidelity monitoring.

[0049] Preferably, the Raman spectroscopy probe monitors the culture sample to obtain the sulfate concentration, hydrogen sulfide concentration and formic acid concentration of the culture sample;

[0050] The ultraviolet spectrophotometer monitors the culture sample to obtain the OD600, nitrate concentration, dissolved organic matter and total organic carbon concentration of the culture sample;

[0051] The water quality sensor monitors the culture sample to obtain one or more of the methane concentration, carbon dioxide concentration, dissolved oxygen concentration, pH value, conductivity, and temperature of the culture sample.

[0052] Preferably, after a single fidelity monitoring is completed, the fidelity monitoring unit is disassembled and cleaned, and after reconnection, the input end corresponding to the second multi-way valve is switched on to perform sampling fidelity monitoring on the remaining pressure-maintaining culture cylinders.

[0053] Compared with the prior art, the technical solution of the present invention has the following beneficial effects:

[0054] The present invention utilizes a gas pressurizing unit to provide high-pressure nitrogen to a pressure-maintaining culture unit and a fidelity monitoring unit, thereby realizing a high-pressure culture environment for deep-sea culture samples; utilizes the gas pressurizing unit to change the nitrogen pressure value injected into a pressure-maintaining culture cylinder and a pressure-maintaining buffer cylinder, thereby realizing pressure-maintaining transfer of deep-sea culture samples among the pressure-maintaining culture cylinder, the pressure-maintaining buffer cylinder and the monitoring compartment; realizes switching sampling of multiple-sequence pressure-maintaining culture cylinders by opening and closing a second multi-way valve, utilizes a single set of fidelity monitoring units to meet the needs of sampling and monitoring of multiple pressure-maintaining culture cylinders, and reduces equipment costs; realizes the whole process of pressure-maintaining operation of sampling and recovering culture samples by opening and closing a three-way valve and a four-way valve, thereby ensuring the authenticity of the culture samples during the monitoring process, reducing the loss of the culture samples during the monitoring process, improving the sampling frequency and efficiency of the culture process, and extending the culture cycle of deep-sea microorganisms. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] Figure 1 This is a schematic diagram of the structure of a multi-sequence fidelity monitoring device for deep-sea microbial culture described in Example 1;

[0056] Figure 2 This is a schematic diagram of the structure of a multi-sequence fidelity monitoring device for deep-sea microbial culture described in Example 2;

[0057] Figure 3 This is a schematic diagram of the structure of the pressure-maintaining culture cylinder described in Example 2;

[0058] Figure 4 This is a schematic structural diagram of the pressure-maintaining buffer cylinder described in Example 2;

[0059] Figure 5 This is a schematic diagram of the structure of the monitoring compartment described in Example 2;

[0060] Figure 6 This is a flow chart of a multi-sequence fidelity monitoring method for deep-sea microbial culture described in Example 3;

[0061] In the figure: 1-gas booster unit, 2-first multi-way valve, 3-pressure-maintaining culture cylinder, 4-second multi-way valve, 5-four-way valve, 6-pressure-maintaining buffer cylinder, 7-monitoring compartment, 8-three-way valve, 9-collection control unit, 10-sampling needle valve, 11-sampling needle valve, 12-pressure sensor, 31-gas sealing end cover, 32-first cavity, 33-first piston, 34-seawater sealing end cover, 61-first sealing end cover, 62-second cavity, 63-second piston, 64-second sealing end cover, 71-third sealing end cover, 72-fourth sealing end cover, 73-cabin, 74-window, 75-Raman spectrometer probe, 76-ultraviolet spectrometer probe, 77-compartment pressure sensor, 78-water quality sensor, 79-liquid level sensor. DETAILED DESCRIPTION

[0062] The drawings are for illustrative purposes only and should not be construed as limiting the present patent;

[0063] In order to better illustrate the present embodiment, some parts in the drawings may be omitted, enlarged or reduced, and do not represent the size of the actual product;

[0064] It is understandable to those skilled in the art that some well-known structures and their descriptions may be omitted in the drawings.

[0065] The technical solution of the present invention is further described below in conjunction with the accompanying drawings and embodiments.

[0066] Example 1

[0067] This embodiment provides a multi-sequence fidelity monitoring device for deep-sea microbial cultivation, such as Figure 1As shown, it includes a gas pressurization unit 1, a pressure-maintaining culture unit, a fidelity monitoring unit and a collection control unit 9;

[0068] The pressure-maintaining culture unit comprises a first multi-way valve 2, a plurality of pressure-maintaining culture cylinders 3 and a second multi-way valve 4; the input end of the first multi-way valve 2 is connected to the first output end of the gas pressurizing unit 1, the plurality of output ends of the first multi-way valve 2 are respectively connected to one end of a pressure-maintaining culture cylinder 3, and the other end of each pressure-maintaining culture cylinder 3 is respectively connected to an input end of the second multi-way valve 4;

[0069] The fidelity monitoring unit includes a three-way valve 8, a four-way valve 5, a pressure-maintaining buffer cylinder 6 and a monitoring compartment 7; the first port of the four-way valve 5 is connected to the output end of the second multi-way valve 4, and the second port of the four-way valve 5 is connected to the input end of the monitoring compartment 7; the third port of the four-way valve 5 is connected to one end of the pressure-maintaining buffer cylinder 6, and the other end of the pressure-maintaining buffer cylinder 6 is connected to the first port of the three-way valve 8; the fourth port of the four-way valve 5 is connected to the second port of the three-way valve 8, and the third port of the three-way valve 8 is connected to the second output end of the gas booster unit 1;

[0070] The input end of the acquisition control unit 9 is connected to the data output end of the monitoring compartment 7, and the output end of the acquisition control unit 9 is respectively connected to the control end of the gas booster unit 1, the control end of the first multi-way valve 2, the control end of the second multi-way valve 4, the control end of the three-way valve 8, and the control end of the four-way valve 5.

[0071] During the specific implementation process, the gas booster unit 1 is used to provide high-pressure nitrogen to the pressure-maintaining culture unit and the fidelity monitoring unit to achieve a high-pressure culture environment for deep-sea culture samples; the gas booster unit 1 realizes the transfer of deep-sea culture samples between the pressure-maintaining culture cylinder 3, the pressure-maintaining buffer cylinder 6 and the monitoring compartment 7 by changing the nitrogen pressure value injected into the pressure-maintaining culture cylinder 3 and the pressure-maintaining buffer cylinder 6; the switching sampling of multi-sequence pressure-maintaining culture cylinders 3 is realized by opening and closing the second multi-way valve 4, and the sampling and monitoring requirements of multiple pressure-maintaining culture cylinders 3 are met by using a single set of fidelity monitoring units, thereby reducing the equipment cost; the whole process of pressure-maintaining operation of sampling and recovery of culture samples is realized by opening and closing the three-way valve 8 and the four-way valve 5, thereby ensuring the authenticity of the culture samples during the monitoring process, reducing the loss of the culture samples during the monitoring process, improving the sampling frequency and efficiency of the culture process, and extending the culture cycle of deep-sea microorganisms.

[0072] Example 2

[0073] This embodiment provides a multi-sequence fidelity monitoring device for deep-sea microbial cultivation, such as Figure 2 As shown, it includes a gas pressurization unit 1, a pressure-maintaining culture unit, a fidelity monitoring unit, a collection control unit 9, a plurality of sampling needle valves 10, a plurality of ventilation needle valves 11 and a plurality of pressure sensors 12;

[0074] The pressure-maintaining culture unit comprises a first multi-way valve 2, a plurality of pressure-maintaining culture cylinders 3 and a second multi-way valve 4; the input end of the first multi-way valve 2 is connected to the first output end of the gas pressurizing unit 1, the plurality of output ends of the first multi-way valve 2 are respectively connected to one end of a pressure-maintaining culture cylinder 3, and the other end of each pressure-maintaining culture cylinder 3 is respectively connected to an input end of the second multi-way valve 4; a ventilation needle valve 11 is arranged between each output end of the first multi-way valve 2 and one end of a pressure-maintaining culture cylinder 3;

[0075] like Figure 3 As shown, each of the pressure-maintaining culture cylinders 3 includes a gas-sealed end cap 31, a first cavity 32, a first piston 33 and a seawater-sealed end cap 34;

[0076] The first piston 33 is disposed inside the first cavity 32, dividing the first cavity 32 into a first gas pressurization cavity and a pressure-maintaining culture cavity; the gas sealing end cover 31 is disposed at one end of the first gas pressurization cavity, and the seawater sealing end cover 34 is disposed at one end of the pressure-maintaining culture cavity;

[0077] The output ends of the first multi-way valve 2 are respectively connected to the gas sealing end cap 31 of a pressure-maintaining culture cylinder 3, and the seawater sealing end cap 34 of each pressure-maintaining culture cylinder 3 is respectively connected to an input end of the second multi-way valve 4;

[0078] The first cavity 32 is a titanium alloy cylindrical structure, with a movable first piston 33 built in. The first piston 33 is sleeved with an O-ring to isolate the first cavity 32 into a first gas pressurization cavity and a pressure-maintaining culture cavity. The gas pressurization unit 1 injects high-pressure nitrogen into the first gas pressurization cavity of the pressure-maintaining culture cylinder 3 through the first multi-way valve 2, thereby realizing a high-pressure culture environment for deep-sea culture samples.

[0079] The fidelity monitoring unit includes a three-way valve 8, a four-way valve 5, a pressure-maintaining buffer cylinder 6 and a monitoring compartment 7; the first port of the four-way valve 5 is connected to the output end of the second multi-way valve 4, and the second port of the four-way valve 5 is connected to the input end of the monitoring compartment 7; the third port of the four-way valve 5 is connected to one end of the pressure-maintaining buffer cylinder 6, and the other end of the pressure-maintaining buffer cylinder 6 is connected to the first port of the three-way valve 8; the fourth port of the four-way valve 5 is connected to the second port of the three-way valve 8, and the third port of the three-way valve 8 is connected to the second output end of the gas booster unit 1 A venting needle valve 11 is arranged between the other end of the pressure-maintaining buffer cylinder 6 and the first port of the three-way valve 8; a sampling needle valve 10 is arranged between the other end of each pressure-maintaining culture cylinder 3 and an input end of the second multi-way valve 4; a sampling needle valve 10 is arranged between the third port of the four-way valve 5 and one end of the pressure-maintaining buffer cylinder 6; a sampling needle valve 10 is arranged between the second port of the four-way valve 5 and the input end of the monitoring compartment 7; each of the pressure sensors 12 is arranged at a sampling end of a sampling needle valve 10;

[0080] like Figure 4 As shown, the pressure-maintaining buffer cylinder 6 includes a first sealing end cover 61, a second cavity 62, a second piston 63 and a second sealing end cover 64;

[0081] The second piston 63 is disposed inside the second cavity 62, dividing the second cavity 62 into a second gas pressurization cavity and a pressure-maintaining buffer cavity; the first sealing end cover 61 is disposed at one end of the pressure-maintaining buffer cavity, and the second sealing end cover 64 is disposed at one end of the second gas pressurization cavity;

[0082] The third port of the four-way valve 5 is connected to the first sealing end cover 61 of the pressure-maintaining buffer cylinder 6, and the second sealing end cover 64 of the pressure-maintaining buffer cylinder 6 is connected to the first port of the three-way valve 8;

[0083] The second cavity 62 is a titanium alloy cylindrical structure, with a movable second piston 63 built in. The second piston 63 is sleeved with an O-ring, dividing the second cavity 62 into a second gas boosting cavity and a pressure maintaining buffer cavity; the gas boosting unit 1 injects high-pressure nitrogen into the second gas boosting cavity of the pressure maintaining buffer cylinder 6 through the three-way valve 8, so as to realize a high-pressure culture environment for deep-sea culture samples; the culture samples in the pressure maintaining culture cylinder 3 enter the pressure maintaining buffer cavity of the pressure maintaining culture cylinder 3 through the second multi-way valve 4 and the four-way valve 5, so as to realize pressure maintaining cache of the culture samples.

[0084] like Figure 5 As shown, the monitoring compartment 7 includes a third sealing end cover 71, a fourth sealing end cover 72, a compartment body 73, a window 74, a Raman spectroscopic probe 75, an ultraviolet spectroscopic probe 76, a compartment pressure sensor 77, a water quality sensor 78 and a liquid level sensor 79;

[0085] The third sealing end cover 71 and the fourth sealing end cover 72 are respectively arranged at the two ends of the cabin body 73, and the viewing window 74 is arranged on the side wall of the cabin body 73; the liquid level sensor 79 is arranged on the lower surface of the third sealing end cover 71, the water quality sensor 78 and the compartment pressure sensor 77 are arranged on the upper surface of the fourth sealing end cover 72, and the Raman spectrometer probe 75 and the ultraviolet spectrometer probe 76 are arranged on the viewing window 74;

[0086] The second port of the four-way valve 5 is connected to the lower surface of the fourth sealing end cover 72 of the monitoring compartment 7;

[0087] The data output ends of the Raman spectroscopic probe 75, the ultraviolet spectroscopic probe 76, the water quality sensor 78 and the liquid level sensor 79 are all connected to the input end of the acquisition control unit 9;

[0088] The cabin 73 is a titanium alloy cylindrical structure, and the fourth sealing end cover 72 is connected to the four-way valve 5 for the injection and outflow of culture samples; the compartment pressure sensor 77 and the liquid level sensor 79 are used to monitor the pressure value and liquid level of the culture samples in the cabin 73 in real time, and are used to reflect the sample quantity in the cabin 73; the Raman spectrometer probe 75, the ultraviolet spectrometer probe 76 and the water quality sensor 78 monitor the various physical and chemical parameters of the culture samples in the cabin 73 in real time; the gas booster unit 1 injects high-pressure nitrogen into the cabin 73 through the three-way valve 8 and the four-way valve 5 to achieve a high-pressure monitoring environment for deep-sea culture samples.

[0089] The water quality sensor 78 includes any one or more of a methane sensor, a carbon dioxide sensor, a dissolved oxygen sensor, a temperature sensor, a conductivity sensor, and a pH sensor; the dissolved oxygen sensor, the temperature sensor, the conductivity sensor, and the pH sensor are all probe sensors;

[0090] The data output end of each of the pressure sensor 12, Raman spectrometer probe 75, ultraviolet spectrometer probe 76, water quality sensor 78 and liquid level sensor 79 is connected to the input end of the acquisition control unit 9, and the output end of the acquisition control unit 9 is respectively connected to the control end of the gas boosting unit 1, the control end of the first multi-way valve 2, the control end of the second multi-way valve 4, the control end of the three-way valve 8, and the control end of the four-way valve 5.

[0091] Example 3

[0092] This embodiment also provides a multi-sequence fidelity monitoring method for deep-sea microbial culture, which is applied to the device described in Example 1 or 2, such as Figure 6 As shown, including:

[0093] S1: The gas pressurization unit is connected to all the pressure-maintaining culture cylinders through the first multi-way valve, connected to the pressure-maintaining buffer cylinder through the three-way valve, connected to the monitoring compartment through the three-way valve and the four-way valve, and all the pressure-maintaining culture cylinders, pressure-maintaining buffer cylinders and monitoring compartments are maintained at the first pressure value through nitrogen pressurization;

[0094] S2: Selecting a pressure-maintaining culture cylinder to be monitored, opening the output end corresponding to the first multi-way valve, and causing the gas pressurizing unit to pressurize the pressure-maintaining culture cylinder to be monitored with nitrogen, so that the pressure of the first gas pressurizing chamber is increased to a second pressure value;

[0095] S3: Open the input end and the output end corresponding to the second multi-way valve, and the first port and the third port of the four-way valve, to connect the pressure-maintaining culture cylinder and the pressure-maintaining buffer cylinder; under the pressure difference between the second pressure value and the first pressure value, the first piston in the pressure-maintaining culture cylinder pushes the culture sample in the pressure-maintaining culture chamber to be injected into the pressure-maintaining buffer chamber of the pressure-maintaining buffer cylinder;

[0096] S4: closing the input end and the output end corresponding to the second multi-way valve and the first port of the four-way valve, opening the first port and the third port of the three-way valve, and the gas boosting unit boosts nitrogen to the pressure-maintaining buffer cylinder to increase the pressure of the second gas boosting chamber to a second pressure value;

[0097] S5: Open the second port of the four-way valve to connect the pressure-maintaining buffer cylinder and the monitoring sub-cabin; under the pressure difference between the second pressure value and the first pressure value, the second piston in the pressure-maintaining buffer cylinder injects the culture sample in the pressure-maintaining buffer cavity into the body of the monitoring sub-cabin;

[0098] S6: When the culture sample reaches the liquid level sensor in the monitoring compartment, the second port and the third port of the four-way valve are closed; the Raman spectroscopic probe, the ultraviolet spectroscopic probe, the compartment pressure sensor, and the water quality sensor monitor the culture sample, and transmit the obtained monitoring results to the acquisition control unit;

[0099] S7: The gas pressurizing unit decompresses the pressure-maintaining buffer cylinder to reduce the pressure of the second gas pressurizing chamber to a third pressure value;

[0100] S8: Open the second port and the third port of the four-way valve to connect the pressure-maintaining buffer cylinder and the monitoring compartment; under the pressure difference between the first pressure value and the third pressure value, the second piston in the pressure-maintaining buffer cylinder sucks the culture sample in the monitoring compartment back to the pressure-maintaining buffer chamber, and close the second port and the third port of the four-way valve;

[0101] S9: The gas pressurizing unit pressurizes the pressure-maintaining buffer cylinder with nitrogen, so that the pressure of the second gas pressurizing chamber is increased to the second pressure value; the gas pressurizing unit depressurizes the pressure-maintaining culture cylinder to be monitored, so that the pressure of the first gas pressurizing chamber is reduced to the first pressure value;

[0102] S10: opening the input end and the output end corresponding to the second multi-way valve, and the first port and the third port of the four-way valve, connecting the pressure-maintaining culture cylinder and the pressure-maintaining buffer cylinder; under the pressure difference between the second pressure value and the first pressure value, the second piston in the pressure-maintaining buffer cylinder pushes the culture sample in the pressure-maintaining buffer cavity to be injected into the pressure-maintaining culture cavity of the pressure-maintaining buffer cavity;

[0103] S11: closing the input end and the output end corresponding to the second multi-way valve, and the first port and the third port of the four-way valve, isolating the pressure-maintaining culture cylinder, and completing single-sequence fidelity monitoring;

[0104] S12: Update the selected pressure-maintaining culture cylinder to be monitored, and repeat steps S2-S11 to achieve multi-sequence fidelity monitoring.

[0105] In the specific implementation process, during the long-term pressure culture of deep-sea culture samples, the pressure value of the pressure sensor in the pressure-maintaining culture unit is checked regularly, and the gas pressurization unit is connected through the first multi-way valve to ensure that the pressure environment of the multi-sequence culture system is maintained at the deep-sea original pressure value, i.e., the first pressure value; during the fidelity monitoring process, the fidelity monitoring unit is connected to the gas pressurization unit through the three-way valve and the four-way valve, and the pressure value of the second gas pressurization chamber of the monitoring compartment and the pressure buffer cylinder is also increased to the first pressure value through nitrogen pressurization. At this time, there is no pressure difference between the pressure-maintaining culture unit and the fidelity monitoring unit.

[0106] A pressure-maintaining culture cylinder to be monitored is selected, and the output end corresponding to the first multi-way valve is opened. The gas boosting unit performs nitrogen boosting on the pressure-maintaining culture cylinder to be monitored, so that the pressure of the first gas boosting chamber is increased to a second pressure value. At the same time, the input end and the output end corresponding to the second multi-way valve, as well as the first port and the third port of the four-way valve are opened to connect the pressure-maintaining culture cylinder and the pressure-maintaining buffer cylinder. Since the pressure value of the pressure-maintaining culture chamber is higher than the pressure value of the pressure-maintaining buffer chamber, the first piston pushes the culture sample in the pressure-maintaining culture chamber into the pressure-maintaining buffer chamber under the action of the pressure difference.

[0107] After the transfer of the culture sample is completed, the input and output ends corresponding to the second multi-way valve and the first port of the four-way valve are closed, and the gas booster unit performs nitrogen boosting to the pressure-maintaining buffer cylinder to increase the pressure of the second gas booster chamber to the second pressure value; the second port of the four-way valve is opened to connect the pressure-maintaining buffer cylinder and the monitoring compartment. Since the pressure value of the pressure-maintaining buffer chamber is higher than the pressure value of the monitoring compartment, the second piston pushes the culture sample in the pressure-maintaining buffer chamber to be injected into the monitoring compartment under the action of the pressure difference; the liquid level sensor is used to determine whether the culture sample is full. After it is full, the second and third ports of the four-way valve are closed to stop the injection. The material of the window of the monitoring compartment is sapphire. The Raman spectroscopic probe outside the window monitors the sulfate concentration, hydrogen sulfide concentration and formic acid concentration of the culture sample. The ultraviolet spectroscopic probe monitors the OD600, nitrate concentration, dissolved organic matter and total organic carbon concentration of the culture sample. The water quality sensor monitors the methane concentration, carbon dioxide concentration, dissolved oxygen concentration, pH value, conductivity and temperature of the culture sample. The data output ends of the water quality sensor, Raman spectroscopic probe and ultraviolet spectroscopic probe are connected to the acquisition control unit to transmit the monitoring results to the acquisition control unit for solution.

[0108] After the fidelity monitoring operation is completed, the culture samples in the monitoring compartment need to be recovered into the pressure-maintaining culture cylinder; the gas booster unit decompresses the pressure-maintaining buffer cylinder to reduce the pressure of the second gas booster chamber to a third pressure value, and opens the second port and the third port of the four-way valve to connect the pressure-maintaining buffer cylinder and the monitoring compartment. Since the pressure value of the monitoring compartment is higher than the pressure value of the pressure-maintaining buffer chamber, the second piston moves under the pressure difference to suck the culture samples in the monitoring compartment back into the pressure-maintaining buffer chamber, and closes the second port and the third port of the four-way valve.

[0109] The gas boosting unit boosts the pressure-maintaining buffer cylinder with nitrogen, so that the pressure of the second gas boosting chamber is increased to the second pressure value. At the same time, the gas boosting unit decompresses the pressure-maintaining culture cylinder to be monitored, so that the pressure of the first gas boosting chamber is reduced to the first pressure value. The input end and the output end corresponding to the second multi-way valve, as well as the first port and the third port of the four-way valve are opened to connect the pressure-maintaining culture cylinder and the pressure-maintaining buffer cylinder. Since the pressure value of the pressure-maintaining buffer chamber is higher than the pressure value of the pressure-maintaining culture chamber, the second piston in the pressure-maintaining buffer cylinder pushes the culture sample in the pressure-maintaining buffer chamber to be injected into the pressure-maintaining culture chamber of the pressure-maintaining buffer chamber. The input end and the output end corresponding to the second multi-way valve, as well as the first port and the third port of the four-way valve are closed to isolate the pressure-maintaining culture cylinder, and word-fidelity monitoring is completed.

[0110] Based on the above sample recovery process, culture medium or nutrients are added to the pressure-maintaining buffer chamber to realize the feeding operation of the pressure-maintaining culture unit; after a single fidelity monitoring is completed, the fidelity monitoring unit is disassembled and cleaned, and after reconnection, the input end corresponding to the second multi-way valve is switched on to perform sampling fidelity monitoring on the remaining pressure-maintaining culture cylinders.

[0111] The method provided in this embodiment can realize the enrichment culture, low-loss fidelity sampling and monitoring of deep-sea microorganisms, improve the authenticity of culture sample monitoring, and low loss can increase the sampling frequency of culture samples and extend the time period of the deep-sea microorganism enrichment culture process.

[0112] The same or similar reference numerals correspond to the same or similar components;

[0113] The terms used in the drawings to describe positional relationships are only used for illustrative purposes and should not be construed as limiting this patent;

[0114] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the embodiments here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. A multi-sequence fidelity monitoring device for deep-sea microbial culture, characterized in that: It comprises a gas pressurizing unit (1), a pressure-maintaining culture unit, a fidelity monitoring unit and a collection control unit (9); The pressure-maintaining culture unit comprises a first multi-way valve (2), a plurality of pressure-maintaining culture cylinders (3) and a second multi-way valve (4); the input end of the first multi-way valve (2) is connected to the first output end of the gas pressurizing unit (1), the plurality of output ends of the first multi-way valve (2) are respectively connected to one end of a pressure-maintaining culture cylinder (3), and the other end of each pressure-maintaining culture cylinder (3) is respectively connected to an input end of the second multi-way valve (4); The fidelity monitoring unit comprises a four-way valve (5), a three-way valve (8), a pressure-maintaining buffer cylinder (6) and a monitoring compartment (7); the first port of the four-way valve (5) is connected to the output end of the second multi-way valve (4), and the second port of the four-way valve (5) is connected to the input end of the monitoring compartment (7); the third port of the four-way valve (5) is connected to one end of the pressure-maintaining buffer cylinder (6), and the other end of the pressure-maintaining buffer cylinder (6) is connected to the first port of the three-way valve (8); the fourth port of the four-way valve (5) is connected to the second port of the three-way valve (8), and the third port of the three-way valve (8) is connected to the second output end of the gas pressurizing unit (1); The input end of the acquisition control unit (9) is connected to the data output end of the monitoring compartment (7), and the output end of the acquisition control unit (9) is respectively connected to the control end of the gas booster unit (1), the control end of the first multi-way valve (2), the control end of the second multi-way valve (4), the control end of the three-way valve (8), and the control end of the four-way valve (5); Each of the pressure-maintaining culture cylinders (3) comprises a gas-sealed end cover (31), a first cavity (32), a first piston (33) and a seawater-sealed end cover (34); The first piston (33) is arranged inside the first cavity (32) to divide the first cavity (32) into a first gas pressurization cavity and a pressure-maintaining culture cavity; the gas sealing end cover (31) is arranged at one end of the first gas pressurization cavity, and the seawater sealing end cover (34) is arranged at one end of the pressure-maintaining culture cavity; The plurality of output ends of the first multi-way valve (2) are respectively connected to a gas-sealed end cover (31) of a pressure-maintaining culture cylinder (3), and the seawater-sealed end cover (34) of each pressure-maintaining culture cylinder (3) is respectively connected to an input end of the second multi-way valve (4); The pressure-maintaining buffer cylinder (6) comprises a first sealing end cover (61), a second cavity (62), a second piston (63) and a second sealing end cover (64); The second piston (63) is arranged inside the second cavity (62) to divide the second cavity (62) into a second gas pressurization cavity and a pressure-maintaining buffer cavity; the first sealing end cover (61) is arranged at one end of the pressure-maintaining buffer cavity, and the second sealing end cover (64) is arranged at one end of the second gas pressurization cavity; The third port of the four-way valve (5) is connected to the first sealing end cover (61) of the pressure-maintaining buffer cylinder (6), and the second sealing end cover (64) of the pressure-maintaining buffer cylinder (6) is connected to the first port of the three-way valve (8).

2. The multi-sequence fidelity monitoring device for deep-sea microbial cultivation according to claim 1, characterized in that: The monitoring compartment (7) comprises a third sealing end cover (71), a fourth sealing end cover (72), a compartment body (73), a viewing window (74), a Raman spectroscopic probe (75), an ultraviolet spectroscopic probe (76), a compartment pressure sensor (77), a water quality sensor (78) and a liquid level sensor (79); The third sealing end cover (71) and the fourth sealing end cover (72) are respectively arranged at two ends of the cabin body (73), and the viewing window (74) is arranged on the side wall of the cabin body (73); the liquid level sensor (79) is arranged on the lower surface of the third sealing end cover (71), the water quality sensor (78) and the compartment pressure sensor (77) are arranged on the upper surface of the fourth sealing end cover (72), and the Raman spectrometer probe (75) and the ultraviolet spectrometer probe (76) are arranged on the viewing window (74); The second port of the four-way valve (5) is connected to the lower surface of the fourth sealing end cover (72) of the monitoring compartment (7); The data output ends of the Raman spectrum probe (75), the ultraviolet spectrum probe (76), the water quality sensor (78) and the liquid level sensor (79) are all connected to the input end of the acquisition control unit (9).

3. The multi-sequence fidelity monitoring device for deep-sea microbial cultivation according to claim 2, characterized in that: The water quality sensor (78) includes any one or more of a methane sensor, a carbon dioxide sensor, a dissolved oxygen sensor, a temperature sensor, a conductivity sensor, and a pH sensor.

4. The multi-sequence fidelity monitoring device for deep-sea microbial cultivation according to claim 3, characterized in that: The dissolved oxygen sensor, temperature sensor, conductivity sensor and pH sensor are all probe-type sensors.

5. The multi-sequence fidelity monitoring device for deep-sea microbial cultivation according to claim 1, characterized in that: The device also includes a plurality of sampling needle valves (10) and a plurality of ventilation needle valves (11); A ventilation needle valve (11) is arranged between each output end of the first multi-way valve (2) and one end of a pressure-maintaining culture cylinder (3); A vent needle valve (11) is provided between the other end of the pressure-maintaining buffer cylinder (6) and the first port of the three-way valve (8); A sampling needle valve (10) is arranged between the other end of each pressure-maintaining culture cylinder (3) and an input end of the second multi-way valve (4); A sampling needle valve (10) is provided between the third port of the four-way valve (5) and one end of the pressure-maintaining buffer cylinder (6); A sampling needle valve (10) is arranged between the second port of the four-way valve (5) and the input end of the monitoring compartment (7).

6. The multi-sequence fidelity monitoring device for deep-sea microbial cultivation according to claim 1, characterized in that: The device also includes a plurality of pressure sensors (12); Each of the pressure sensors (12) is correspondingly arranged at a sampling end of a sampling needle valve (10), and a data output end of each of the pressure sensors (12) is connected to an input end of a collection control unit (9).

7. A multi-sequence fidelity monitoring method for deep-sea microbial culture, applied to the device of claim 2, characterized in that: include: S1: The gas pressurizing unit (1) is connected to all the pressure-maintaining culture cylinders (3) through the first multi-way valve (2), connected to the pressure-maintaining buffer cylinder (6) through the three-way valve (8), and connected to the monitoring compartment (7) through the three-way valve (8) and the four-way valve (5), and all the pressure-maintaining culture cylinders (3), the pressure-maintaining buffer cylinder (6) and the monitoring compartment (7) are maintained at the first pressure value through nitrogen pressurization; S2: selecting a pressure-maintaining culture cylinder (3) to be monitored, opening the output end corresponding to the first multi-way valve (2), and causing the gas pressurizing unit (1) to pressurize the pressure-maintaining culture cylinder (3) to be monitored with nitrogen, so that the pressure of the first gas pressurizing chamber is increased to a second pressure value; S3: opening the input end and the output end corresponding to the second multi-way valve (4), and the first port and the third port of the four-way valve (5), so as to connect the pressure-maintaining culture cylinder (3) and the pressure-maintaining buffer cylinder (6); under the pressure difference between the second pressure value and the first pressure value, the first piston (33) in the pressure-maintaining culture cylinder (3) pushes the culture sample in the pressure-maintaining culture chamber to be injected into the pressure-maintaining buffer chamber of the pressure-maintaining buffer cylinder (6); S4: the input end and the output end corresponding to the second multi-way valve (4) and the first port of the four-way valve (5) are closed, the first port and the third port of the three-way valve (8) are opened, and the gas boosting unit (1) boosts the nitrogen pressure in the pressure-maintaining buffer cylinder (6), so that the pressure of the second gas boosting chamber is increased to a second pressure value; S5: opening the second port of the four-way valve (5) to connect the pressure-maintaining buffer cylinder (6) and the monitoring sub-cabin (7); under the pressure difference between the second pressure value and the first pressure value, the second piston (63) in the pressure-maintaining buffer cylinder (6) injects the culture sample in the pressure-maintaining buffer cavity into the cabin (73) of the monitoring sub-cabin (7); S6: When the culture sample reaches the liquid level sensor (79) in the monitoring compartment (7), the second port and the third port of the four-way valve (5) are closed; the Raman spectroscopic probe (75), the ultraviolet spectroscopic probe (76), the compartment pressure sensor (77), and the water quality sensor (78) monitor the culture sample, and transmit the obtained monitoring results to the acquisition control unit (9); S7: the gas pressurizing unit (1) decompresses the pressure-maintaining buffer cylinder (6), so that the pressure of the second gas pressurizing chamber is reduced to a third pressure value; S8: opening the second port and the third port of the four-way valve (5) to connect the pressure-maintaining buffer cylinder (6) and the monitoring compartment (7); under the pressure difference between the first pressure value and the third pressure value, the second piston (63) in the pressure-maintaining buffer cylinder (6) sucks the culture sample in the monitoring compartment (7) back to the pressure-maintaining buffer chamber, and closing the second port and the third port of the four-way valve (5); S9: the gas pressurizing unit (1) pressurizes the pressure-maintaining buffer cylinder (6) with nitrogen, so that the pressure of the second gas pressurizing chamber is increased to the second pressure value; the gas pressurizing unit (1) depressurizes the pressure-maintaining culture cylinder (3) to be monitored, so that the pressure of the first gas pressurizing chamber is reduced to the first pressure value; S10: opening the input end and the output end corresponding to the second multi-way valve (4), and the first port and the third port of the four-way valve (5), so as to connect the pressure-maintaining culture cylinder (3) and the pressure-maintaining buffer cylinder (6); under the pressure difference between the second pressure value and the first pressure value, the second piston (63) in the pressure-maintaining buffer cylinder (6) pushes the culture sample in the pressure-maintaining buffer cavity to be injected into the pressure-maintaining culture cavity of the pressure-maintaining culture cylinder (3); S11: closing the input end and the output end corresponding to the second multi-way valve (4), and the first port and the third port of the four-way valve (5), isolating the pressure-maintaining culture cylinder (3), and completing single-sequence fidelity monitoring; S12: Update the selected pressure-maintaining culture cylinder (3) to be monitored, and repeat steps S2-S11 to achieve multi-sequence fidelity monitoring.

8. The multi-sequence fidelity monitoring method for deep-sea microbial culture according to claim 7, characterized in that: The Raman spectroscopy probe (75) monitors the culture sample to obtain the sulfate concentration, hydrogen sulfide concentration and formic acid concentration of the culture sample; The ultraviolet spectrophotometer (76) monitors the culture sample to obtain the OD600, nitrate concentration, dissolved organic matter and total organic carbon concentration of the culture sample; The water quality sensor (78) monitors the culture sample to obtain one or more of the methane concentration, carbon dioxide concentration, dissolved oxygen concentration, pH value, conductivity, and temperature of the culture sample.

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