WO2023072309A1

WO2023072309A1 - Intelligent solution injection device for deep-sea sediment microorganism culture experiment

Info

Publication number: WO2023072309A1
Application number: PCT/CN2022/138890
Authority: WO
Inventors: 李洁; 陈煜�; 张偲
Original assignee: 南方海洋科学与工程广东省实验室(广州); 中国科学院南海海洋研究所
Priority date: 2021-12-28
Filing date: 2022-12-14
Publication date: 2023-05-04
Also published as: CN114540178B; CN114540178A

Abstract

Disclosed in the present invention is an intelligent solution injection device for a deep-sea sediment microorganism culture experiment. The intelligent solution injection device is mainly composed of an insertion device, a controllable slow-release solution injection system, an interstitial water displacement and monitoring system, and a deep-sea peristaltic pump. The insertion device can insert a solution injection needle tube and an interstitial water collection tube into a designated sediment position by means of gravity or other driving devices; the controllable slow-release solution injection system is used for receiving an instruction to inject a certain amount of culture solution into sediment through the solution injection needle tube; and the interstitial water displacement and monitoring system is used for collecting sediment interstitial water and monitoring environmental parameters to develop a solution injection policy. The present invention can simulate a diffusion process of matter seeping out of a deep-sea cold seep, and can provide a powerful tool for researchers to conduct long-term in-situ experiments of deep-sea sediment microorganisms.

Description

An intelligent liquid injection device for deep-sea sediment microbial culture experiments

Technical field:

The invention relates to the field of deep-sea sediment microbial cultivation experiments, in particular to an intelligent liquid injection device for deep-sea sediment microbial cultivation experiments.

Background technique:

The current microbial scientific research on deep-sea sediments is based on sampling and analysis, and what is studied is only the microbial state at a certain point in time. Due to the lack of in-situ experimental equipment, the inability to obtain time-series data of deep-sea submarine life activities, and the difficulty in conducting research in frontier fields such as the response of deep-sea organisms to environmental changes, the current progress in deep-sea life science is slow.

Single-factor environmental change experiment is a common method in life science research. Scientists need to maintain the concentration of a substance in seabed sediments continuously to observe the life activity process of microorganisms and the law of ecological succession. Patent document CN105758687A discloses a deep-sea sediment in-situ cultivation and sealed sampling mechanism, but it only injects the nutrient solution into the deep-sea sediment once for the cultivation of microorganisms, and then takes the samples back after a certain time point. An experiment with changing environment has been carried out, but it still cannot solve the problems of maintaining the experimental conditions and collecting the time series data required for the study of life activities.

Invention content:

The object of the present invention is to provide an intelligent liquid injection device for deep-sea sediment microbial cultivation experiments that can be mounted on a deep-sea in-situ experimental platform.

In order to achieve the above purpose, the intelligent liquid injection device designed by the present invention is mainly composed of an insertion device, a controllable slow-release liquid injection system, a gap water replacement monitoring system and a deep-sea peristaltic pump; the intelligent liquid injection device can collect sediment gaps Water and monitor environmental data, judge according to the set conditions and send instructions to automatically control the amount of nutrient solution injection and the injection time interval to maintain the concentration of nutrient solution in the sediment required for in-situ long-term culture experiments of deep-sea microorganisms; the insertion device can pass gravity or other driving devices to insert the injection needle and interstitial water collection tube into the designated sediment position; the controllable slow-release injection system is used to receive instructions to inject a certain amount of culture solution into the sediment through the injection needle; interstitial water The displacement monitoring system is used to collect sediment interstitial water and monitor environmental parameters to formulate injection strategies.

As an improvement of the present invention, the controllable slow-release liquid injection system includes a motor control mechanism, a vertical deceleration movement mechanism, a guiding and pressing guide rod, a liquid movement protection cabin and a liquid sealing cabin; the motor control mechanism is used to drive the vertical deceleration The moving mechanism drives the guide rod to do linear reciprocating motion; the liquid-tight compartment is fixed, and the opening end is sealed by a flexible cup-shaped rubber sleeve; the liquid movement protection cabin is connected to the guide rod and the cup-shaped rubber sleeve; the liquid moves Driven by the guiding rod, the protective cabin presses the cup-shaped rubber sleeve into the liquid-sealed compartment, so that the culture medium in the liquid-sealed compartment flows out through the outlet of the one-way valve at the bottom. The edge of the cup-shaped rubber sleeve has a semicircular convex The structural design makes the upper and lower cabins sealed when they are in contact with each other, and one end of the sealed cabin is provided with a flange that is more than 3 times the radius of curvature of the cup-shaped rubber sleeve to make it move when it is folded to fit the bulkhead.

As an improvement of the present invention, the liquid-tight compartment is designed with an upper circle and a lower conical surface, so that the liquid injection is complete and more efficient; it is made of transparent materials; it is made of acid-resistant, alkali-resistant and corrosion-resistant materials, which can meet various liquid requirements storage, so that the experiment has more ways.

As an improvement of the present invention, the insertion device includes a liquid injection needle tube, a liquid injection port, a gap water collection pipe, a liquid suction port, and a guiding and pressing column; a number of outlets for upward concave pressure are arranged at intervals on the liquid injection needle tube. A nozzle, so that when the injection needle tube is inserted into the sediment, the sediment will not enter the inside of the injection needle tube; the gap water collection tube includes a gap water filter membrane, a collection shell and a fixed inner shell; the collection shell is connected to the fixed inner shell, and the gap water The water filtration membrane is set in the interlayer between the collection shell and the fixed inner shell, and the shell of the gap water collection shell and the gap water collection fixed inner shell where the gap water filtration membrane is located are all provided with a plurality of water holes, from top to bottom The number of lower water holes increases gradually, and the gap between holes gradually decreases. This structure can uniformly collect sediment interstitial water at different depths.

As an improvement of the present invention, the liquid injection needle tube and the interstitial water collection fixed inner shell are fixed on the guiding and pressing column, and the guiding and pressing column is connected with the mobile device of the deep-sea in-situ experimental platform, which can be controlled by gravity or other forces. Driven, effectively inserted into the sediment; the liquid injection port is located at the top of the liquid injection needle tube, and is connected with the one-way valve outlet of the liquid-tight chamber of the controllable slow-release liquid injection system through a pipe; the liquid suction port is located at the top of the interstitial water collection fixed inner shell , and connected to the water inlet of the deep-sea peristaltic pump through a pipeline.

As an improvement of the present invention, the interstitial water replacement monitoring system includes a deep-sea peristaltic pump, a interstitial water cabin, a cabin sealing cover, a sensor, a water inlet, a water outlet, and a control circuit; the deep-sea peristaltic pump slowly inserts the interstitial water in the device It is sucked into the fixed cabin of the sensor, and the original seawater in the cabin is replaced and discharged. The flow rate of the deep-sea peristaltic pump is not more than 100ml/min; the cabin sealing cover is airtightly connected with the interstitial water cabin, and the interior is filled with seawater. The water inlet is set on the sealing cover of the cabin The bottom is connected to the water outlet of the deep-sea peristaltic pump; the water outlet is set on the top of the side of the interstitial water compartment away from the compartment sealing cover, and communicates with the outside in one direction; the sensor is fixed on the bottom of the compartment sealing cover and the interstitial water compartment; the control system It is sealed with high pressure resistance, connected with the sensor through a watertight connector, accepts the sensor data and stores it in a self-contained manner, and at the same time judges whether it is lower than the set lower limit and feeds back the output to the motor control mechanism of the controllable slow-release injection system to control the start of the motor. stop.

Compared with prior art, the beneficial effect of the present invention is:

1. The intelligent liquid injection device of the present invention realizes an intelligent control device for long-term cultivation experiments of deep-sea microorganisms. By regularly collecting interstitial water in sediments, monitoring the injection concentration and detecting the culture status of microorganisms, the obtained data are used to analyze the environmental conditions of the culture experiment, judge the threshold value and formulate the injection strategy, and feed back to the motor control mechanism to automatically adjust the injection time And the amount of injection, to maintain the concentration of substances required for microbial culture experiments in sediments for a long time.

2. The intelligent liquid injection device of the present invention inserts the liquid injection needle into the sediment through the drive of the experimental platform, and the liquid injection range is not inside the sediment sampling tube, and the natural diffusion of the injected liquid in the sediment can simulate the deep sea Diffusion process of substances in cold seeps.

3. The intelligent liquid injection device of the present invention can realize the long-term culture of microorganisms under single-factor stable control in the natural state of deep-sea in-situ sediment microorganisms, and provide effective tools for scientific researchers to conduct deep-sea in-situ experiments.

Description of drawings

Fig. 1 is a schematic diagram of the overall structure of the intelligent liquid injection device.

Fig. 2 is a schematic diagram of the structure of the controllable slow-release injection system.

3A and 3B are schematic diagrams of the structure of the insertion device.

Fig. 4 is a schematic structural diagram of the interstitial water displacement detection system.

Fig. 5 is a structural schematic diagram of a deep-sea peristaltic pump;

Explanation of reference signs: 1-controllable slow-release injection system; 2-insertion device; 3-interstitial water displacement detection system; 4-deep sea peristaltic pump;

1.1-Motor control mechanism; 1.2 Nut screw moving mechanism; 1.3-Guide and press down guide rod; 1.4-Liquid movement protection cabin; 1.5-Liquid sealed cabin;

2.1- Liquid injection port; 2.2 Liquid injection needle tube; 2.3 Interstitial water filtration membrane; 2.4- Interstitial water collection shell; 2.5- Interstitial water collection fixed inner shell; 2.6- Suction port;

3.1-Cabin sealing cover; 3.2-Sensor; 3.3-Water inlet; 3.4-Cabin body; 3.5-Water outlet;

4.1-Water inlet of deep-sea peristaltic pump; 4.2-Water outlet of deep-sea peristaltic pump.

Detailed ways:

In order to make the object, technical scheme and effect of the present invention more clear and definite, the present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.

Example

As shown in Figures 1 to 5, an intelligent liquid injection device for deep-sea sediment microbial cultivation experiments that can be mounted on a deep-sea in-situ experimental platform mainly consists of a controllable slow-release liquid injection system 1, an insertion device 2, Interstitial water displacement detection system 3 and deep sea peristaltic pump 4. The insertion device 2 inserts it into the designated sediment position and required depth through gravity or other driving devices. When the platform becomes stable, the controllable slow-release injection system 1 starts to work, slowly injecting the cultured liquid into the sediment . After a certain period of time, the deep-sea peristaltic pump 4 starts to work and sucks the interstitial water penetrated into the insertion device 2 into the interstitial water replacement detection system 3, and discharges the original seawater in the cavity through the water flow movement in the system cavity. The sensors in the interstitial water replacement detection system 3 detect various parameters in the interstitial water, and feed back relevant data to the controllable slow-release liquid injection system 1 to control the amount of liquid injection and the time interval.

The controllable slow-release liquid injection system 1 is mainly composed of a motor control mechanism 1.1, a nut screw moving mechanism 1.2, a guiding and pressing guide rod 1.3, a liquid moving protection cabin 1.4, and a liquid sealing cabin 1.5. The motor control mechanism 1.1 is equipped with a brushless motor with an encoder, a star reducer and a worm gear reducer that saves vertical space, and a sensor data feedback control module and a current detection module are added inside. Through these The module controls the start and stop of the motor and the power-off action at the end of liquid injection. The screw rod of the nut screw moving mechanism 1.2 is fixedly connected to both ends of the cabin body through ceramic bearings, one end of which is connected to the turbine through a flat key, and the fixed plane at the nut end is connected to the fixed plate guiding the downward pressure guide rod 1.3. The vertical downward movement of the rod 1.3 places the fixed plate on it in the square channel of the cabin body, limiting its degree of freedom of rotation. The guiding and pressing guide rod 1.3 has a pair of symmetrical guide rods connected non-fixedly with the fixed nut in the middle of the liquid movement protection cabin 1.4, which facilitates the installation and disassembly of the controllable slow-release liquid injection system 1 after filling the liquid. In order to increase the amount of liquid injection, two liquid movement protection cabins 1.4 are provided, and the fixed sleeve on them is fixed to the support plate of the experimental platform. One side of the cabin body is provided with an opening to ensure that the guide rod and the liquid guide rod 1.3 are guided down. The guide sleeve of mobile protection cabin 1.4 moves up and down. The liquid-tight compartment 1.5 isolates the liquid from the outside world through a soft cup-shaped rubber sleeve, and prevents the seawater from entering the outside world through a one-way valve. The volume space of the cap when it moves to the maximum position is determined by the one-way valve that the liquid-sealed cabin 1.5 communicates with the outside world. Since the liquid-sealed cabin 1.5 is under water with internal and external pressure balance, in order to ensure the controllable slow-release injection The weight of the system 1 is made of transparent PC, and when the liquid is loaded, the liquid filling situation of the sealed compartment can be observed with the naked eye.

The insertion device 2 is mainly composed of a liquid injection port 2.1, a liquid injection needle tube 2.2, an interstitial water filter membrane 2.3, an interstitial water collection shell 2.4, an interstitial water collection fixed inner shell 2.5, a liquid suction port 2.6 and a guiding and pressing column 2.7. The liquid injection port 2.1 is connected with the two one-way valve outlets in the liquid-tight compartment 1.5 of the controllable slow-release liquid injection system 1, so that the liquid is injected into the sediment by the liquid injection needle 2.2 through this port. When the liquid flows out, set two upward concave water outlets (as shown in Figure 3B) at the required positions of the tubular structure of the injection needle 2.2, so that when the sediment is pressed into the deposit, it will not cause the sediment to enter the liquid injection needle 2.2 . The interstitial water filtration membrane 2.3 is arranged in the interlayer between the interstitial water collection shell 2.4 and the interstitial water collection fixed inner shell 2.5, preventing the insertion device 2 from entering the cavity along the water hole on the shell during the process of inserting the sediment, Affecting the suction of interstitial water and the size of the inner space of the cavity, a plurality of water holes are set on the housing of the interstitial water collection shell 2.4 and the interstitial water collection fixed inner case 2.5; at this time, the liquid injection needle tube 2.2 and the interstitial water collection fixed The inner shell 2.5 is connected with the guiding and pressing column 2.7 by welding, and the guiding and pressing column 2.7 is inserted into the sediment through the moving device of the deep-sea in-situ experiment platform. The pumping port 2.6 is connected with the deep-sea peristaltic pump water inlet 4.1 of the deep-sea peristaltic pump 4 .

The interstitial water displacement detection system 3 is mainly composed of a cabin sealing cover 3.1, a sensor 3.2, a water inlet 3.3, a cabin body 3.4 and a water outlet 3.5. The interstitial water replacement detection system 3 sucks the interstitial water into the cavity of the system 3 through the water inlet 3.3 through the suction of the deep-sea peristaltic pump 4. In order to fully fill the interstitial water in the cavity, the water inlet 3.3 is placed at the bottom of the system 3. The water outlet 3.5 is set above the bottom of the detection system 3, the sensor 3.2 is fixed and sealed on the bottom of the cabin sealing cover 3.1 and the cabin body 3.4, and the relevant detected data is fed back to the motor control mechanism 1.1 in the controllable slow-release injection system 1 , to control the start and stop of the motor.

The intelligent liquid injection device of this application has an independent insertion device and a controllable slow-release liquid injection mechanism; at the same time, an in-situ interstitial water-related data acquisition system is designed. On the one hand, it feeds back to the liquid injection mechanism to control the liquid injection time and Liquid ratio, to maintain the concentration of substances required for the experiment, on the other hand, the time series of environmental and microbial state data can be obtained.

At the same time, this application uses a controllable slow-release liquid injection system to accurately inject the liquid required for the experiment into the sediment at a specified depth. The liquid is inserted independently without the corresponding resistance of the viscosity of the sediment during the injection process. The device is not limited by other sampling devices, and accurately simulates the material diffusion process when the deep-sea cold seep seeps out.

The above-mentioned embodiments are only to illustrate the technical concept and characteristics of the present invention, and its purpose is to enable those of ordinary skill in the art to understand the content of the present invention and implement it accordingly, and cannot limit the protection scope of the present invention. All equivalent changes or modifications made according to the essence of the present invention shall fall within the protection scope of the present invention.

Claims

An intelligent liquid injection device for deep-sea sediment microbial in-situ culture experiments, characterized in that it is mainly composed of an insertion device, a controllable slow-release liquid injection system, a gap water replacement monitoring system, and a deep-sea peristaltic pump. The water monitoring data automatically adjusts the nutrient solution injection amount and injection time interval, so as to maintain the nutrient solution concentration in the sediment required for the in-situ long-term culture experiment of deep-sea microorganisms; the insertion device can move the injection needle and the gap through gravity or other driving devices The water collection tube is inserted into the designated sediment position; the controllable slow-release injection system is used to receive instructions to inject a certain amount of culture solution into the sediment through the injection needle; the interstitial water displacement monitoring system is used to collect interstitial water in the sediment And monitor environmental parameters to formulate injection strategies.
The intelligent liquid injection device according to claim 1, characterized in that: the controllable slow-release liquid injection system includes a motor control mechanism, a vertical deceleration movement mechanism, a guiding and pressing down guide rod, a liquid movement protection cabin and a liquid sealing cabin; The motor control mechanism is used to drive the vertical deceleration moving mechanism to drive the guide down-press guide rod to do linear reciprocating motion; the liquid-tight cabin is fixed, and the opening end is sealed by a flexible cup-shaped rubber sleeve; the liquid movement protection cabin is respectively connected to the guide-down guide rod and The cup-shaped rubber sleeve is connected; the liquid movement protection cabin is driven by the guide rod to press the cup-shaped rubber sleeve into the liquid-sealed compartment, so that the culture medium in the liquid-sealed compartment flows out through the outlet of the one-way valve at the bottom; the cup-shaped rubber The edge of the sleeve has a semicircular raised structure designed to seal the upper and lower cabins when they are in contact with each other. One end of the bulkhead sealed compartment is provided with a flange that is more than 3 times the curvature radius of the cup-shaped rubber sleeve.
The intelligent liquid injection device according to claim 2, wherein the liquid-tight chamber is designed with an upper circle and a lower cone surface, and is made of acid-resistant, alkali-resistant, and corrosion-resistant materials.
The intelligent liquid injection device according to claim 1, characterized in that: the insertion device includes a liquid injection needle, a liquid injection port, a gap water collection pipe, a liquid suction port, and a guiding column; the upper interval of the liquid injection needle is A number of upwardly concave water outlets are provided, so that when the liquid injection needle is inserted into the sediment, the sediment will not enter the interior of the liquid injection needle; the gap water collection tube includes a gap water filter membrane, a collection shell and a fixed inner shell, and the collection shell Connected to the fixed inner shell, the interstitial water filtration membrane is arranged in the interlayer of the collection shell and the fixed inner shell, and the gap water collection shell where the interstitial water filtration membrane is located and the shell of the gap water collection fixed inner shell are provided with multiple The number of water holes increases from top to bottom, and the gap between holes gradually decreases.
The intelligent liquid injection device according to claim 4, characterized in that: the liquid injection needle tube and the interstitial water collection fixed inner shell are fixed on the guiding and pressing column, and the guiding and pressing column is connected with the mobile device of the deep-sea in-situ experimental platform; Note The liquid port is located on the top of the liquid injection needle, and is connected to the outlet of the one-way valve of the liquid-tight chamber of the controllable slow-release liquid injection system through a pipe; Water outlet connection.
The intelligent liquid injection device according to claim 1, characterized in that: the gap water replacement monitoring system includes a gap water cabin, a cabin sealing cover, a sensor, a water inlet, a water outlet, and a control system; the deep-sea peristaltic pump will be inserted into the device The interstitial water in the sensor is slowly sucked into the fixed cabin of the sensor, and the original seawater in the cabin is replaced and discharged. The flow rate of the deep sea peristaltic pump is not more than 100ml/min; The bottom of the cabin sealing cover is connected with the water outlet of the deep-sea peristaltic pump; the water outlet is set on the top of the side of the interstitial water compartment away from the compartment sealing cover, and communicates with the outside in one direction; the sensor is fixed on the compartment sealing cover and the interstitial water compartment Bottom: The control system is sealed with high pressure resistance, connected to the sensor through a watertight connector, accepts the sensor data and stores it in a self-contained manner, and at the same time judges whether it is lower than the set lower limit and feeds back to the motor control mechanism of the controllable slow-release injection system, Control the start and stop of the motor.