Deep-sea microorganism enrichment culture and mineral reaction integrated device

Abstract

The invention discloses a deep-sea microorganism enrichment culture and mineral reaction integrated device, belongs to the field of microorganism equipment, and aims to solve the problems that deep-sea extreme microorganism resource development is limited and mineral reaction research and observation are difficult. A metabolite and an enzyme system of the strain have remarkable application value, but a traditional enrichment method is limited by extreme environments such as high pressure, low temperature and oligotrophic environments, and efficient development is difficult to realize; in addition, deep sea mineral reaction related research is blocked due to difficult direct observation and lack of in-situ observation equipment. The device comprises a microorganism enrichment culture device and a mineral reaction device, the microorganism enrichment culture device is used for suction filtration and enrichment of deep sea microorganism samples and can collect in-situ water samples; the mineral reaction device is used for researching the interaction between the deep sea extreme environment and a mineral sample.

Description

Technical Field

[0001] This invention relates to the field of microbial equipment technology, specifically to an integrated device for deep-sea microbial enrichment culture and mineral reaction. Background Technology

[0002] The deep sea is rich in extremophile microorganisms, whose metabolites and unique enzyme systems have irreplaceable applications in industrial catalysis, new drug development, and other fields. The enrichment of deep-sea microorganisms is the result of the combined effects of extreme environmental pressures, nutrient distribution, and biological interactions. This reflects both the "survival wisdom" of microorganisms adapting to the deep sea and is a core process for maintaining deep-sea ecological balance and global geochemical cycles. With the continuous development of deep-sea exploration technology, in-depth analysis of this phenomenon will not only reveal the mechanisms of extreme adaptation in life but also provide a new perspective for resource development and environmental regulation.

[0003] However, deep-sea microorganisms live in extreme environments such as high pressure, low temperature, and low nutrition, resulting in extremely low population densities in their natural state. Traditional enrichment methods face numerous challenges. Conventional laboratory cultures cannot simulate in-situ high-pressure environments, leading to the inhibition of barotropic microbial activity; insufficient nutrient substrate compatibility prevents the precise activation of functional groups such as chemosynthetic bacteria; and imbalanced competition among dominant populations during enrichment can easily lead to the loss of target microorganisms. Existing technologies mostly rely on static pressurization devices or single nutrient additions, resulting in low enrichment efficiency, poor community fidelity, and insufficient recovery rates of target strains. These issues severely restrict the development and utilization of deep-sea microbial resources.

[0004] Therefore, exploring mineral reactions and microbial enrichment under extreme deep-sea environments is of great significance for research on deep-sea mineral development, global climate change and element cycling, and deep-sea ecosystems. Deep-sea mineral reactions refer to the chemical, physical, and biochemical interactions between minerals and surrounding water, fluids, rocks, or organisms in the unique deep-sea environment (high pressure, low temperature / localized high temperature, high salinity, darkness, rich fluids, etc.). Driven by the unique environmental conditions of the deep sea, these reactions not only shape the distribution and morphology of deep-sea minerals but also profoundly participate in global element cycling, support deep-sea ecosystems, and form important mineral resources, especially in areas such as hydrothermal vent areas, cold seep areas, and deep-sea polymetallic nodule mining areas.

[0005] However, the extreme nature of the deep-sea environment makes direct observation extremely difficult. Currently, our understanding of mineral reactions largely relies on laboratory simulations, and we lack the necessary equipment for in-situ observation of deep-sea mineral reactions. Summary of the Invention

[0006] To address the shortcomings of existing technologies, this invention proposes an integrated device for deep-sea microbial enrichment culture and mineral reaction, characterized by comprising a microbial enrichment culture device and a mineral reaction device; The microbial enrichment and culture device is used for filtering and enriching deep-sea microbial samples and can collect in-situ water samples. The mineral reaction device is used to study the interaction between the extreme deep-sea environment and mineral samples.

[0007] As a further improvement of the present invention, the microbial enrichment and culture device includes a first battery and a control chamber and at least three microbial enrichment modules. Each microbial enrichment module is connected to a valve group unit via a connecting pipeline. The valve group unit is sequentially connected to a multi-channel valve body and a water pump via a hydraulic pipeline.

[0008] As a further improvement of the present invention, the valve group unit includes a 2-position solenoid valve and a second 4-position solenoid valve.

[0009] As a further improvement of the present invention, the microbial enrichment module includes a water sampling bottle, the upper end of which is connected to a one-way valve, and the lower end of which is connected to one end of a flow meter via a connecting pipe. The other end of the flow meter is connected to a microbial enrichment bottle via a pipe, which contains three layers of microbial filter membranes for microbial filtration. A first 4-position solenoid valve is installed on the connecting pipe. A fixed liquid bottle is provided between the water sampling bottle and the microbial enrichment bottle. The fixed liquid bottle is connected to the microbial enrichment bottle via a pipe that passes through the first 4-position solenoid valve and the flow meter in sequence.

[0010] As a further improvement of the present invention, the microbial enrichment bottle includes a first enrichment bottle body, a second enrichment bottle body, a third enrichment bottle body, a fourth enrichment bottle body, and a fifth enrichment bottle body that are interconnected. The lower end of the fifth enrichment bottle body is provided with a multi-channel valve interface. The protrusion of the first enrichment bottle body is provided with a water sampling pipeline interface and a fixative pipeline interface. The top of the protrusion of the first enrichment bottle body is provided with a water injection cap.

[0011] As a further improvement of the present invention, the mineral reaction device includes a support frame, with linear bearings around the support frame and a motor compartment connected to its top. A second battery and a control compartment are connected above the motor compartment. A motor is sealed inside the motor compartment. A reaction tank with an open top is provided on the inner end face of the support frame. A reaction tank cover is fixedly installed below the motor compartment and corresponds to the position of the reaction tank. A mineral basket is connected to the lower end of the reaction tank cover and can be placed inside the reaction tank. A rubber bladder is provided on one side of the reaction tank. Guide columns are provided around the support frame.

[0012] As a further improvement of the present invention, the lifting mechanism is a trapezoidal lead screw and a lead screw nut; One end of the trapezoidal lead screw is connected to the motor output shaft in the motor compartment, and the other end is fixedly connected to the pressure plate of the reaction vessel through the lead screw nut and the pressure plate sealing O-ring. The lower end of the trapezoidal lead screw is provided with a lead screw protective sleeve and a lead screw sealing O-ring.

[0013] The beneficial effects of this invention are: 1. Design a deep-sea mineral reaction device to fill the gap in deep-sea mineral reaction equipment.

[0014] 2. All parts of the equipment body that come into contact with seawater are made of non-metallic materials to avoid interference from the materials of the equipment body with the mineral reaction and microbial extraction process.

[0015] 3. Before and after the in-situ reaction of deep-sea minerals, the minerals, microorganisms, and collected water samples are all sealed to prevent seawater exchange during equipment deployment and recovery from interfering with the reaction results.

[0016] 4. Multiple reaction devices can be deployed simultaneously to conduct time-series observations and study the mineral reactions and microbial distribution status in different time periods.

[0017] 5. The microbial enrichment device can collect water while performing biological enrichment, and retain the water sample during the enrichment process.

[0018] 6. While achieving water collection and bioaccumulation in multiple modules, each module is sealed and isolated from the others to avoid interference.

[0019] 7. By comparing the results of microbial and mineral reactions obtained by filtration at the same time, the organic influence of microbial species and content on mineral reactions was studied, and the interaction mechanism between the two was investigated. Attached Figure Description

[0020] Figure 1 This is a structural diagram of the microbial enrichment and culture device in this embodiment; Figure 2 This is a structural diagram of the microbial enrichment module in this embodiment; Figure 3 This is a structural diagram of the valve assembly unit in this embodiment; Figure 4 This is a structural diagram of the microbial enrichment bottle in this embodiment; Figure 5 This is a structural diagram of the mineral reaction device in this embodiment, wherein (a) is a structural diagram of the mineral reaction device when the lifting mechanism is in the raised state, and (b) is a structural diagram of the mineral reaction device when the lifting mechanism is in the lowered state; Figure 6 This is an enlarged sectional view of the lifting mechanism; Figure 7 This is a structural diagram of the mineral basket in this embodiment.

[0021] In the diagram: 1. First battery and control compartment; 2. Microbial enrichment module; 3. Valve unit; 4. Multi-channel valve body; 5. Water pump; 201. One-way valve; 202. Water sampling bottle; 203. First 4-position solenoid valve; 204. Flow meter; 205. Microbial enrichment bottle; 206. Fixative bottle; 207. Piping; 2051. Water sampling pipe interface; 2052. Fixative pipe interface; 2053. First enrichment bottle; 2054. Second enrichment bottle; 2055. Third enrichment bottle; 2056. Fourth enrichment bottle; 2057. ... Five enrichment bottles, 2058, multi-channel valve interface, 2059, water injection cap, 301, 2-position solenoid valve, 302, second 4-position solenoid valve, 101, second battery and control compartment, 102, motor compartment, 103, trapezoidal lead screw, 104, guide column, 105, linear bearing, 106, lead screw nut, 107, rubber bladder, 108, mineral basket, 109, cap sealing O-ring, 1010, reaction vessel cap, 1011, bracket, 1012, reaction vessel, 1013, lead screw protective sleeve, 1014, lead screw sealing O-ring. Detailed Implementation

[0022] To facilitate understanding of this application, a more comprehensive description will be provided below with reference to the accompanying drawings. Embodiments of this application are illustrated in the drawings. These embodiments are provided to provide a more thorough and complete understanding of the disclosure of this application.

[0023] To facilitate understanding of this application, a more comprehensive description will be provided below with reference to the accompanying drawings. Embodiments of this application are illustrated in the drawings. Rather, these embodiments are provided to provide a more thorough and complete understanding of the disclosure of this application.

[0024] In this embodiment, to study the relationship between microbial changes and mineral reactions in the deep-sea environment, a time series of microbial enrichment and mineral reactions was established. Specifically, filtration was performed on designated enrichment bottles and mineral reaction units simultaneously. By comparing the results of the microbial samples obtained from filtration and the mineral reaction, the specific impact of microbial species and content on the mineral reaction was explored, thereby investigating the mechanism of action between the two. In addition, the integrated device for deep-sea microbial enrichment culture and mineral reaction can be tested on the seabed using an unmanned remotely operated vehicle (ROV) or carried on a deep-sea lander.

[0025] like Figures 1-7 As shown, an integrated device for deep-sea microbial enrichment culture and mineral reaction includes a microbial enrichment culture device and a mineral reaction device. The microbial enrichment and culture device is used for filtering and enriching deep-sea microbial samples and can collect in-situ water samples. The mineral reaction device is used to study the interaction between the extreme deep-sea environment and mineral samples.

[0026] The microbial enrichment and culture device includes a first battery and control chamber 1 for sealing the battery and control system devices, and at least three microbial enrichment modules 2. Each microbial enrichment module 2 is connected to a valve group unit 3 via a connecting pipe 207. The valve group unit 3 is sequentially connected to a multi-channel valve body 4 and a water pump 5 via a hydraulic pipe. The multi-channel valve body 4 merges the pipes of multiple microbial enrichment modules 2 together and connects to a single outlet connected to the water pump 5. The water pump 5 is used to filter seawater. The microbial enrichment module 2 is used for water sampling and microbial enrichment. It includes a water sampling bottle 202 for storing seawater samples during filtration. The upper end of the water sampling bottle 202 is connected to a one-way valve 201 to control the unidirectional flow of the filtered seawater. The lower end of the water sampling bottle 202 is connected to one end of a flow meter 204 via a connecting pipe. The other end of the flow meter 204 is connected to a microbial enrichment bottle 205 via a pipe 207. The microbial enrichment bottle 205 contains three layers of microbial filter membranes for microbial filtration. The flow meter 204 counts the volume of filtered seawater. A first 4-position solenoid valve 203 is installed on the connecting pipe. A fixative bottle 206 is provided between the water sampling bottle 202 and the microbial enrichment bottle 205. The fixative bottle 206 is connected to the microbial enrichment bottle 205 via a pipe that passes through the first 4-position solenoid valve 203 and the flow meter 204 in sequence. The first 4-position solenoid valve 203 is used to control the opening and closing of the pipe and control the extraction of seawater and fixative. The fixative can fix the microorganisms present on the filter membrane.

[0027] The microbial enrichment bottle 205 includes a first enrichment bottle 2053, a second enrichment bottle 2054, a third enrichment bottle 2055, a fourth enrichment bottle 2056, and a fifth enrichment bottle 2057 that are interconnected. Each enrichment bottle is sealed with an O-ring. Each of the first enrichment bottle 2053, second enrichment bottle 2054, third enrichment bottle 2055, and fourth enrichment bottle 2056 is equipped with three layers of filter membranes. These three microbial filter membranes are of three different sizes: 5 micrometers, 0.45 micrometers, and 0.22 micrometers. After the microbial enrichment culture device reaches the designated water depth, the microbial enrichment bottle 205 operates according to a set time sequence of the water pump, pumping water to allow deep-sea seawater to pass through the filter membranes. This filters bacteria and other microorganisms from the deep-sea seawater onto the filter membranes. Then, the water pump extracts a fixative solution, which passes through the filter membrane to fix the filtered microorganisms on the membrane, preventing microbial movement over a long period and ensuring accurate statistical results. After all filtration is completed, the recovery device is brought to the water surface, and the filter membrane is removed for microbial statistics.

[0028] The fifth enrichment bottle 2057 has a multi-channel valve interface 2058 at its lower end for connecting to the pipeline leading to the multi-channel valve. The protrusion of the first enrichment bottle 2053 has a water sampling pipeline interface 2051 and a fixed liquid pipeline interface 2052. The top of the protrusion of the first enrichment bottle 2053 is provided with a water injection cap 2059 for filling the equipment with pure water during the initial preparation of the equipment.

[0029] The microbial enrichment unit has a total of 6 enrichment pipelines, which are controlled by 6 solenoid valves.

[0030] The microbial enrichment unit consists of three microbial enrichment modules 2, one valve group unit 3, one multi-channel valve body 4, one water pump 5, a first battery, and a control chamber 1.

[0031] Each microbial enrichment module 2 has two microbial enrichment bottles 205 and two water sampling bottles 202, with the water sampling bottles 202 and microbial enrichment bottles 205 connected in series, allowing for water sampling while enrichment is being performed. Every two enrichment bottles share one fixed liquid bottle 206, and the connection between the fixed liquid bottle and the two enrichment bottles is controlled by two solenoid valves. Each module has one 4-position solenoid valve compartment.

[0032] Taking a single microbial enrichment bottle 205 as an example, the specific usage method is as follows: Before the equipment is lowered, pure water is injected into all pipelines in the system by combining water injection with water pumping. The filtration time for each enrichment bottle is set. After the equipment is deployed to the seabed, when the working time of the microbial enrichment bottle 205 begins, the 2-position solenoid valve 301 and the 4-position solenoid valve 302 are opened. After opening, the water pump 5 pumps water to carry out bioaccumulation in the microbial enrichment bottle 205 connected by pipelines and to collect water at the same time. After the water pump 5 has been working for the specified time, the enrichment of microbial enrichment bottle 205 is completed. The 2-position solenoid valve 301 is closed, and the 4-position solenoid valve 302 is opened to extract the fixative. After the fixative is extracted for the specified time, the 2-position solenoid valve 301 and the 4-position solenoid valve 302 are closed to complete the bioaccumulation of the first group.

[0033] The working process of each microbial enrichment bottle 205 in the three microbial enrichment modules 2 is the same as that of the microbial enrichment bottle 205 mentioned above. Each microbial enrichment bottle 205 performs filtration at a specified working time to achieve temporal enrichment of microorganisms.

[0034] The mineral reaction device includes a support 1011, with linear bearings 105 arranged around its perimeter. A motor compartment 102 is connected to the top of the support 1011. A second battery and a control compartment 101 are connected above the motor compartment 102. A motor is sealed inside the motor compartment 102. A reaction tank 1012 with an open top is arranged on the inner end face of the support 1011. A reaction tank cover 1010 is fixedly installed below the motor compartment 102 and corresponds to the position of the reaction tank 1012. A mineral basket 108 is connected to the lower end of the reaction tank cover 1010 and can be placed inside the reaction tank 1012. A rubber bladder 107 is arranged on one side of the reaction tank 1012. Guide columns 104 are arranged around the support 1011.

[0035] A lifting mechanism is fixed on the pressure cap 1010 of the reaction vessel, which is a trapezoidal lead screw 103 and a lead screw nut 106. One end of the trapezoidal lead screw 103 is connected to the motor output shaft in the motor compartment 102, and the other end is fixedly connected to the pressure plate of the reaction vessel 1010 through the lead screw nut 106 and the pressure plate sealing O-ring 109. The lower end of the trapezoidal lead screw 103 is provided with a lead screw sealing O-ring 1014 and a lead screw protective sleeve 1013.

[0036] The second battery and control compartment 101 is sealed with a battery and control system to provide power and control for the device; the motor compartment 102 is sealed with a stepper motor to drive the trapezoidal lead screw 103.

[0037] Before deployment, the stepper motor drives the trapezoidal lead screw 103 to rotate a certain number of times, and the lead screw nut 106 moves upward a certain distance. Since the lead screw nut 106 is fixedly connected to the reaction vessel cover 1010, and the reaction vessel cover 1010 is fixedly connected to the mineral basket 108, the mineral basket 108 can be lifted.

[0038] After the mineral basket 108 is raised, the minerals are placed into the mineral basket 108, and pure water is filled into the reaction vessel 1012. Then, the stepper motor is controlled to drive the trapezoidal screw to rotate a certain number of times, causing the reaction vessel cover 1010 to drop a certain distance, so that the cover sealing O-ring 109 on the reaction vessel cover 1010 engages with the reaction vessel 1012 to seal the mineral basket 108.

[0039] After setting the start and end times of the device, the mineral reaction device is deployed on the seabed.

[0040] When the working time arrives, the stepper motor drives the trapezoidal lead screw 103 to rotate a certain number of times, lifting the reaction tank cover 1010, so that the minerals in the mineral basket 108 are fully exposed to the seabed water environment to carry out mineral reaction. The mineral basket 108 has 3 placement compartments, which can hold multiple pieces of minerals.

[0041] After the end of the working time, the stepper motor drives the trapezoidal lead screw 103 to rotate a certain number of revolutions, lowering the reaction vessel cover 1010 so that the O-ring 109 on the reaction vessel cover 1010 engages with the reaction vessel 1012, sealing the mineral basket 108. Afterwards, the mineral reaction apparatus is recovered. After recovery, the reaction vessel cover 1010 is raised to obtain the reacted minerals.

[0042] Among them, the guide column 104 and the linear bearing 105 play a guiding role in the up and down movement of the reaction vessel cover 1010.

[0043] A rubber bladder 107 is sealed to the reaction vessel 1012 via an O-ring. The rubber bladder 107 balances the pressure inside and outside the reaction vessel 1012, and its elasticity provides a buffer space for water inside the reaction vessel 1012 when the reaction vessel gland 1010 is lowered and the gland sealing O-ring 109 is sealed. A screw rod protective sleeve 1013 is sealed to the reaction vessel gland 1010 via a screw rod sealing O-ring 1014 to prevent water at the screw rod from entering the reaction vessel 1012 and interfering with the mineral reaction.

[0044] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. An integrated device for deep-sea microbial enrichment culture and mineral reaction, characterized in that, The device comprises a microorganism enrichment culture device and a mineral reaction device. The microorganism enrichment culture device is used for filtering and enriching deep-sea microorganism samples and can collect in-situ water samples. The mineral reaction device is used for studying the interaction between deep-sea extreme environment and mineral samples.

2. The apparatus according to claim 1, wherein the apparatus is characterized by: The microorganism enrichment culture device comprises a first battery and control cabin (1) and at least three microorganism enrichment modules (2), the microorganism enrichment modules (2) are connected with a valve group unit (3) through a connecting pipeline (207), and the valve group unit (3) is sequentially connected with a multi-channel valve body (4) and a water pump (5) through a hydraulic pipeline.

3. The apparatus according to claim 2, wherein the apparatus is characterized by: The valve group unit (3) comprises a 2-position electromagnetic valve (301) and a second 4-position electromagnetic valve (302).

4. The apparatus according to claim 3, wherein the apparatus is characterized by: The microorganism enrichment module (2) comprises a water sampling bottle (202), the upper end of the water sampling bottle (202) is connected with a one-way valve (201), the bottom end of the water sampling bottle (202) is connected with one end of a flowmeter (204) through a connecting pipeline, the other end of the flowmeter (204) is connected with a microorganism enrichment bottle (205) through a pipeline (207), the microorganism enrichment bottle (205) contains three layers of microorganism filter membranes and is used for filtering microorganisms, a first 4-position electromagnetic valve (203) is installed on the connecting pipeline, and a fixing liquid bottle (206) is arranged between the water sampling bottle (202) and the microorganism enrichment bottle (205).

5. The apparatus according to claim 4, wherein the apparatus is characterized by: The microorganism enrichment bottle (205) comprises a first enrichment bottle body (2053), a second enrichment bottle body (2054), a third enrichment bottle body (2055), a fourth enrichment bottle body (2056) and a fifth enrichment bottle body (2057) which are in communication with each other, the lower end of the fifth enrichment bottle body (2057) is provided with a multi-channel valve interface (2058), the convex part of the first enrichment bottle body (2053) is provided with a water sampling pipeline interface (2051) and a fixing liquid pipeline interface (2052), and the top end of the convex part of the first enrichment bottle body (2053) is provided with a water injection gland (2059).

6. The apparatus according to claim 1, wherein the apparatus is characterized by: The mineral reaction device comprises a support (1011), linear bearings (105) are arranged around the support (1011), a motor cabin (102) is connected to the top end of the support (1011), a second battery and control cabin (101) is connected to the upper side of the motor cabin (102), a motor is sealed in the motor cabin (102), a reaction tank (1012) with an open upper end is arranged on the inner end face of the support (1011), a reaction tank gland (1010) is fixedly installed below the motor cabin (102) and corresponds to the position of the reaction tank (1012), a mineral basket (108) is connected to the lower end of the reaction tank gland (1010) and can be placed in the reaction tank (1012), a rubber bladder (107) is arranged on one side of the reaction tank (1012), and guide columns (104) are arranged around the support (1011).

7. The apparatus according to claim 6, wherein the apparatus is characterized by: The reaction tank gland (1010) is fixed with a lifting mechanism.

8. The apparatus according to claim 7, wherein the apparatus is characterized by: The lifting mechanism is a trapezoidal screw rod (103) and a screw nut (106); One end of the trapezoidal screw rod (103) is in transmission connection with a motor output shaft in a motor cabin (102), the other end is in fixed connection with a reaction tank gland (1010) through the screw nut (106) and an O-shaped ring (109) sealing gland, and the lower end of the trapezoidal screw rod (103) is provided with a screw rod protection sleeve (1013) and a screw rod sealing O-shaped ring (1014).

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