Device and method for synchronously measuring metabolic rate and excretion rate of aquatic organisms

Abstract

According to the aquatic organism metabolism rate and excretion rate synchronous measurement device and method, on one hand, a metabolism bin, a water pumping bin and related measurement circulation equipment of a middle layer metabolism measurement module are matched with the design of a top layer cover plate, synchronous measurement of the metabolism rate and the excretion rate is achieved, the defects of traditional step-by-step measurement can be avoided, and the experiment efficiency is improved; therefore, the data has better timeliness and relevance, and the physiological state of aquatic organisms is truly and accurately reflected. On the other hand, the temperature of the sealed heating bin can be accurately controlled, the biological stress reaction is reduced, and the stability of the measurement environment is ensured; the arrangement of the water pumping port and the rubber ball is used for balancing the pressure inside and outside the bin during water pumping, and the connection and sealing design of the box body and the top cover plate is used for maintaining the overall sealing performance of the device and ensuring the stability of the measurement environment. In addition, an aquatic organism limiting device is arranged in the metabolism bin, the metabolism measurement space can be flexibly adjusted according to the size and specification of an organism, and the metabolism rate measurement accuracy can be improved by limiting the organism activity and reducing the organism activity frequency.

Description

Technical Field

[0001] The present application relates to the technical field of aquatic organism metabolism measurement, and in particular to a device and method for synchronously measuring the metabolic rate and excretion rate of aquatic organisms. Background Art

[0002] The metabolic rate and excretion rate of aquatic organisms are important indicators for evaluating their physiological state and environmental adaptability, and are also one of the important contents of bioenergetics research. The metabolic rate mainly reflects the energy consumption of organisms. It can reveal the energy allocation strategy of organisms under different environmental pressures and the survival mechanism under limited resource conditions. It is of great significance for evaluating the energy balance of organisms under specific environmental conditions. The excretion rate describes the ability of organisms to eliminate metabolic waste (especially ammonia metabolites), which is important for understanding how organisms maintain internal environmental homeostasis in different ecological environments. By studying the interaction between metabolism and excretion and various biological or abiotic factors, it can provide a scientific basis for ecological research, biological monitoring and ecosystem management of aquatic organisms. Through the determination of metabolic rate and excretion rate, researchers can evaluate the impact of environmental changes on organisms, predict the adaptability of organisms to environmental pressures, and formulate corresponding protection and management measures, thereby promoting the sustainable development of aquatic ecosystems.

[0003] With the development of science and technology, the methods for determining metabolic rate have been constantly evolving, mainly including direct calorimetry, indirect calorimetry and formula estimation. In the field of aquatic biological research, a method is generally used to determine metabolic rate, that is, aquatic organisms (shellfish, shrimp and crabs) are placed in a closed respiration chamber, and the oxygen concentration in the respiration chamber is accurately measured at different time points. The oxygen consumption at each time point is calculated, and finally a mathematical model is used to scientifically estimate the metabolic rate of the organism. The operating procedures for the excretion rate of aquatic organisms are similar to those for the determination of metabolic rate, and the organisms also need to be placed in a specific container. However, the difference is that the determination of excretion rate does not require the container to be in a closed state. By extracting water samples from the containers at the beginning and end of the experiment, and determining the initial and final ammonia nitrogen concentrations in the water according to the sodium hypobromite oxidation method, the excretion rate of the organism can be accurately calculated.

[0004] It can be seen from the above-mentioned metabolic rate and excretion rate determination methods that both determination methods require aquatic organisms to be placed in specific measurement containers. However, for most aquatic organisms, changing different environments each time they are tested will cause stress responses in the organisms. Stress response, as a natural physiological response of an organism to changes in the external environment, will quickly trigger a series of physiological changes in the organism, including but not limited to a rapid increase in stress hormone levels, impaired immune system, etc. These physiological changes will have a significant impact on the normal metabolic process and excretion mechanism of the organism, causing the metabolic rate and excretion rate determination results of the organism to deviate from the actual level, ultimately affecting the accuracy and reliability of the experiment. At present, the determination of the metabolic rate and excretion rate of aquatic organisms is usually carried out in steps. Such experimental operations not only prolong the experimental cycle, but also easily cause stress responses in the organism, affecting the accuracy and reliability of the experimental data. In addition, the currently commonly used method for determining the metabolic rate and excretion rate is to place the organism directly in a closed test bottle without restricting its activities. However, when performing standard metabolic rate determination, the prerequisite is that the organism is required to remain in a static state, otherwise it will lead to increased differences between experimental individuals, affecting the accuracy of the test results. Summary of the invention

[0005] In view of this, the present application provides a device and method for synchronously measuring the metabolic rate and excretion rate of aquatic organisms, which can synchronously measure the metabolic rate and excretion rate of aquatic organisms in a resting state to obtain accurate and reliable experimental data.

[0006] Specifically, the present application is implemented through the following technical solutions: In a first aspect, the present application provides a device for synchronously measuring the metabolic rate and excretion rate of an aquatic organism, the device comprising a box body, in which an intermediate layer metabolic measurement module is arranged; The intermediate layer metabolic measurement module comprises a metabolic chamber and a water pumping chamber which are arranged adjacent to each other, and the metabolic chamber and the water pumping chamber are separated by a first partition plate; An aquatic organism limiting device is provided at the upper part of the metabolic chamber, and the aquatic organism limiting device is communicated with the water flow inside the metabolic chamber; The bottom of the aquatic organism limiting device is provided with a plurality of slots for adjusting in the horizontal direction, and a breathing bottle is arranged above the slots; a plurality of second partition plates are arranged at intervals in the slots, and the height of the second partition plates is higher than that of the breathing bottle, and the side surface of each second partition plate is in contact with the breathing bottle; the size of the space separated by the breathing bottle and the corresponding second partition plates is the same as the size of the aquatic organism to be tested, and the separated space is used to place the aquatic organism to be tested; a plurality of hollow holes are arranged on the breathing bottle and the second partition plates, and the separated space is connected with the outside of the aquatic organism limiting device through the plurality of hollow holes.

[0007] The second aspect of the present application provides a method for synchronously measuring the metabolic rate and excretion rate of an aquatic organism, the method comprising: After water is injected into the intermediate layer metabolism determination module, an aquatic organism limiting device is placed in the metabolism chamber, and the aquatic organism is placed in the breathing bottle; After the oxygen exposure operation, and when the dissolved oxygen concentration reaches the preset threshold, the box is sealed with the top cover; After a specified period of time, the oxygen consumption rate is measured by the dissolved oxygen meter probe, and water is pumped through the water extraction port to calculate the excretion rate.

[0008] The present application provides a device and method for synchronously measuring the metabolic rate and excretion rate of aquatic organisms. An aquatic organism limiting device is provided in the upper part of the metabolic chamber, a horizontally adjustable slot is provided at the bottom of the aquatic organism limiting device, a breathing bottle is provided above the slot, and the height of the slot is higher than the breathing bottle, and the side of the second partition plate is in contact with the side of the breathing bottle, so that a space with the same size as the aquatic organism to be measured can be accurately separated. In this way, the swimming behavior of aquatic organisms of different sizes can be effectively restricted, so that they can maintain a resting state during the measurement process, and the accuracy of metabolic rate measurement can be improved. In addition, the multiple hollow holes on the breathing bottle and the second partition plate connect the separated space with the outside, and while restricting the biological activities, the smooth flow of the internal circulating water is guaranteed, which helps to maintain the environmental stability of the entire metabolic chamber, reduce the stress response caused by environmental fluctuations, and ensure that the dissolved oxygen concentration of the water in various places inside the metabolic module is consistent, so that the measured metabolic rate and excretion rate are closer to the real physiological level of the organism. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1 An exploded view schematic diagram of a device for synchronously measuring the metabolic rate and excretion rate of aquatic organisms provided in the present application; Figure 2 A top view of the intermediate metabolism module shown in this application; Figure 3 A schematic diagram of the structure of the aquatic organism limiting device provided for this application; Figure 4 This is a schematic diagram of the structure of the intermediate layer metabolism measurement module shown in this application; Figure 5 A ventral view of the top lid shown for this application; Figure 6 A front view of the top cover shown for this application; Figure 7 A flow chart of the method for simultaneously determining the metabolic rate and excretion rate of aquatic organisms provided in this application; Description of reference numerals: 1-Dissolved oxygen meter probe access hole; 2-Water outlet; 3-Top cover plate; 4-Spiral hole; 5-Water pumping port; 6-Rubber ball binding hole; 7-Gasket placement track; 8-Box; 9-Heating rod placement slot; 10-Metabolic chamber; 11-Breathing bottle; 12-Heating chamber; 13-Water pumping chamber; 14-Circulating pump, 15-Card slot, 16-Second partition plate, 17-Water inlet, 18-First partition plate, 19-Bacterial oxygen consumption chamber. DETAILED DESCRIPTION

[0010] Here, exemplary embodiments are described in detail, and examples thereof are shown in the accompanying drawings. When the following description refers to the drawings, unless otherwise indicated, the same numbers in different drawings represent the same or similar elements. The implementations described in the following exemplary embodiments do not represent all implementations consistent with the present application.

[0011] The terms used in this application are only for the purpose of describing specific embodiments and are not intended to limit this application. The singular forms of "a", "said" and "the" used in this application are also intended to include plural forms, unless the context clearly indicates other meanings. It should also be understood that the term "and / or" used in this article refers to and includes any or all possible combinations of one or more associated listed items.

[0012] It should be understood that although the terms first, second, third, etc. may be used in the present application to describe various information, these information should not be limited to these terms. These terms are only used to distinguish the same type of information from each other. For example, without departing from the scope of the present application, the first information may also be referred to as the second information, and similarly, the second information may also be referred to as the first information. Depending on the context, the word "if" as used herein may be interpreted as "at the time of" or "when" or "in response to determining".

[0013] Specific embodiments are given below to introduce the technical solution of the present application in detail.

[0014] Figure 1 This is a schematic diagram of an exploded view of a device for synchronously measuring the metabolic rate and excretion rate of aquatic organisms provided in this application. Figure 2 This is a top view of the intermediate metabolism module shown in this application (the bacterial oxygen consumption chamber is not shown). Please also refer to Figure 1 and Figure 2 , the device provided in this embodiment includes a box body 8, in which an intermediate layer metabolism measurement module is provided; The intermediate layer metabolic measurement module includes a metabolic chamber 10 and a water pumping chamber 13 which are arranged adjacent to each other, and the metabolic chamber 10 and the water pumping chamber 13 are separated by a first partition plate 18; An aquatic organism limiting device is provided at the upper part of the metabolic chamber 10, and the aquatic organism limiting device is connected to the water flow inside the metabolic chamber; Figure 3 For the schematic diagram of the aquatic organism limit device provided for this application, please refer to Figure 3 The bottom of the aquatic organism limiting device has a plurality of slots 15 that can be adjusted in the horizontal direction, and a breathing bottle 11 is arranged above the slots 15; a plurality of second partition plates 16 are arranged in intervals in the slots 15, and the height of the second partition plates 16 is higher than the breathing bottle 11, and the side of each second partition plate 16 is in contact with the breathing bottle 11; the size of the space separated by the breathing bottle 11 and the corresponding second partition plates 16 is the same as the size of the aquatic organism to be tested, and the separated space is used to place the aquatic organism to be tested; a plurality of hollow holes are provided on the breathing bottle 11 and the second partition plates 16, and the separated space is connected to the outside of the aquatic organism limiting device through the plurality of hollow holes.

[0015] Please refer to Figure 2 A circulation pump 14 is provided on the side of the first partition plate 18. The circulation pump 14 is located below the aquatic organism limiting device. The circulation pump 14 drives the water flow inside the intermediate layer metabolism measurement module to circulate.

[0016] It should be noted that the metabolic chamber 10 and the pumping chamber 13 are arranged adjacent to each other and separated by the first partition 18. This layout makes the two relatively independent and interrelated in function. The metabolic chamber 10 is mainly used to accommodate aquatic organisms for metabolic rate measurement related operations, while the pumping chamber 13 is used to extract water samples to measure the excretion rate. The presence of the first partition 18 ensures that when the pumping operation is performed, there will be no excessive interference with the aquatic organisms and the water environment in the metabolic chamber 10. At the same time, it also provides a suitable position for the installation of the circulation pump 14, so that the water in the metabolic chamber 10 can flow to the pumping chamber 13 in an orderly manner under the action of the circulation pump 14, so as to realize the circulation of water in the entire intermediate layer metabolic measurement module, ensure the stability of the water environment and the uniformity of the internal dissolved oxygen during the experiment, and help to improve the accuracy of the measurement of metabolic rate and excretion rate.

[0017] The aquatic organism limiting device is arranged at the upper part of the metabolic chamber 10, and is connected to the water flow inside the metabolic chamber 10, which ensures that the aquatic organisms can be in a normal water flow environment while being restricted in their activities, and maintain the relative stability of their physiological state. Among them, the main function of the aquatic organism limiting device is to accurately limit aquatic organisms of different specifications, so that they remain in a resting state during the test, reduce the metabolic differences between individuals caused by biological activities, and thus improve the accuracy of metabolic rate determination. Specifically, different specifications can be limited by a device designed to be pullable, or designed by a slider, sliding to different places to represent different body lengths, widths and heights of aquatic organisms.

[0018] Please continue to refer to Figure 2 The bottom of the aquatic organism limiting device has multiple slots 15 for horizontal adjustment, which can make the entire aquatic organism limiting device flexible to adjust according to different experimental requirements and the specifications of the aquatic organisms to be tested. By moving the slots 15, the relative positions of the breathing bottle 11 and the second partition plate 16 can be changed to adapt to aquatic organisms of different sizes and achieve accurate positioning of organisms of various specifications.

[0019] Among them, a breathing bottle 11 is arranged above the card slot 15, and the shape of each section thereof can be a square structure (or other shapes, which can be set according to actual needs), and the hollow design between the top and the two sides of the breathing bottle 11 facilitates the flow of internal circulating water. In addition, the breathing bottle 11 not only provides a relatively independent living space for aquatic organisms, but also works together with the second partition plate 16 to separate a space of the same size as the aquatic organism to be measured, which can effectively limit the swimming behavior of aquatic organisms and prevent the organisms from excessive activity during the measurement process.

[0020] In addition, the second partition plates 16 disposed in the slots 15 are all higher than the breathing bottles 11, and the side surfaces of each second partition plate 16 are in close contact with the breathing bottles 11. Here, the second partition plates 16 play a role of precise isolation and positioning, and can form a space matching the size of the organisms together with the breathing bottles 11. Its height design ensures that aquatic organisms will not escape from above, and the close contact of the side surfaces prevents the aquatic organisms from moving in the horizontal direction, further enhancing the limiting effect on the aquatic organisms.

[0021] It should also be noted that the multiple hollow holes on the breathing bottle 11 and the second partition plate 16 allow the separated space to be connected to the outside of the aquatic organism limiting device through these holes. In this way, on the one hand, the hollow holes ensure that the water flow can flow evenly throughout the metabolic chamber 10, so that oxygen, nutrients, etc. can be evenly distributed in each breathing bottle 11, and it is also beneficial to the discharge of metabolic waste. On the other hand, the circulation of water helps to maintain a constant water temperature and reduce the impact of local temperature changes on organisms. For aquatic organisms, stable water flow and a suitable water quality environment can reduce stress responses and enable them to maintain a relatively stable physiological state during the measurement process, thereby improving the accuracy and reliability of the experimental results.

[0022] Please continue to refer to Figure 1The device also includes two cylindrical channels (water pumping port 5 and rubber ball binding hole 6) arranged in parallel. The first port of the cylindrical channel is arranged inside the water pumping bin, and the second port is arranged outside the box body, which is used to connect the inside of the water pumping bin 13 and the outside of the box body 8; wherein one second port is connected to the water pumping device (water pumping port 5), and the other second port is connected to the rubber ball (rubber ball binding hole 6) near one side of the water pumping bin. It should be noted that such a layout establishes a direct communication path between the inside of the water pumping bin 13 and the outside of the box body 8, providing a convenient channel structure for operations such as water sample extraction and pressure balance.

[0023] It should be noted that the first port of one of the cylindrical channels is disposed inside the pumping tank 13, and the second port is connected to the pumping device (the pumping port 5), so that water samples can be conveniently drawn from the pumping tank 13 during the experiment. Through the pumping device, a certain amount of water samples can be accurately obtained according to a predetermined time interval or experimental requirements, which is used for the subsequent determination of the change in the concentration of ammonia nitrogen in the water body, thereby calculating the excretion rate of aquatic organisms.

[0024] The first port of another cylindrical channel is arranged inside the water pumping bin 13, and the second port is connected to the rubber ball (rubber ball binding hole 6). The purpose of this is to ensure that the partial pressure inside and outside the water body is consistent when the water flow in the metabolic measurement module is extracted. When the water sample in the water pumping bin 13 is extracted using the water pumping device, the pressure in the bin will change as the internal water flow is extracted. If there is no pressure balancing measure, it may cause difficulty and inaccuracy in water sample extraction, and may even have adverse effects on aquatic organisms and experimental environment in the device. The existence of the rubber ball is used to block the entry of external air. When pumping water, the pressure inside the bin will decrease, causing external air to enter. If the entry of external air is not blocked, it will affect the oxygen content in the water body in the bin, resulting in inaccurate biological metabolic rate data measured. Specifically, the rubber ball can block the entry of external air, and deform accordingly according to the change of pressure in the bin, play a role in buffering and regulating pressure, and ensure that the pressure inside and outside the water body is always balanced during the entire pumping process, so that the pumping operation can be carried out smoothly, while ensuring the stability of the experimental environment, which is conducive to obtaining accurate and reliable excretion rate measurement results.

[0025] It should be noted that by setting up two cylindrical channels (respectively, the water pumping port 5 and the rubber ball binding hole 6), while ensuring the normal circulation of water in the metabolic chamber and the aquatic organisms in a suitable state for metabolic rate determination, water samples can be extracted in a timely and accurate manner for excretion rate determination. By connecting the two ports of the water pumping device and the rubber ball, the effective determination of excretion rate related data is achieved without interfering with the metabolic rate determination process, thereby achieving the simultaneous conduct of the two determinations in the same device, improving the experimental efficiency, and reducing the errors and biological stress reactions that may be caused by step-by-step operations.

[0026] It should also be noted that the intermediate layer metabolic determination module is also provided with a bacterial oxygen consumption chamber 19 on the pumping chamber side. Figure 4 This is a schematic diagram of the structure of the intermediate layer metabolic determination module shown in this application, please refer to Figure 4 The bacterial oxygen consumption bin 19 on the pumping bin side is sealed, and has an inlet and outlet and a dissolved oxygen meter probe access hole (not shown in the figure). The bacterial oxygen consumption bin is used to measure the total oxygen consumption of bacteria in the water body during the metabolic process. Specifically, the water sample in the bacterial oxygen consumption bin 19 is replaced through the inlet and outlet (or a related pump, etc.) at the beginning or end of the test. After the replacement, the sealed state is maintained, and the oxygen consumption of bacteria and other microorganisms in the water sample is measured by the dissolved oxygen meter probe. In conventional experiments, for convenience, when calculating the oxygen consumption rate, the actual oxygen consumption of aquatic organisms needs to be subtracted from the oxygen consumption of bacteria in the water body, and the oxygen consumption of bacteria is represented by the concentration difference of dissolved oxygen in the water body in the bacterial oxygen consumption bin at the end of the experiment and the beginning of the experiment. Since the water needs to be changed regularly according to the experimental requirements when measuring the metabolism of aquatic organisms, and the bacterial content in the water body replaced each time is inconsistent, the bacterial oxygen consumption is inconsistent during each experiment, which will eventually increase the individual differences in the actual oxygen consumption of aquatic organisms and affect the accurate determination of the metabolic rate. The present application sets up a bacterial oxygen consumption bin. Every time the water is replaced, the water in the bacterial oxygen consumption bin is replaced synchronously, and then the bacterial oxygen consumption in the bacterial oxygen consumption bin is measured. The total bacterial oxygen consumption in the metabolic bin and the pumping bin can be calculated based on the volume ratio of the water in the bacterial oxygen consumption bin and the metabolic bin and the pumping bin, so that the final biological oxygen consumption calculation is more accurate.

[0027] Please continue to refer to Figure 2 There is a water flow interlayer between the box body 8 and the metabolic measurement module, and the water flow interlayer is sealed to form a heating chamber 12. In addition, a heating rod placement groove 9 is provided on the side of the heating chamber 12 close to the outside of the box body 8, and a heating rod is provided in the heating rod placement groove 9, and the heating rod heats the water in the water flow interlayer.

[0028] It should be noted that the water flow interlayer between the box body 8 and the metabolic measurement module forms a heating chamber 12. The sealing design of the water flow interlayer effectively prevents internal water leakage and heat exchange with the external environment. The sealed water flow interlayer can accommodate a certain amount of water, which plays a key role in the entire heating process as a medium for heat transfer. The heating rod placement groove 9 provided on the side of the heating chamber 12 close to the outside of the box body 8 provides a stable and suitable installation position for the heating rod. The design of the heating rod placement groove 9 ensures that the heating rod can fully contact the water in the water flow interlayer, thereby efficiently transferring heat to the water body.

[0029] Specifically, the heating rod generates heat by doing work through electric current, and the heat generated is transferred to the water in the water flow sandwich. Since water has a high specific heat capacity, it can absorb a large amount of heat while its own temperature rises relatively slowly, which makes the heating process more uniform and avoids the occurrence of local overheating. By controlling the power and working time of the heating rod, the temperature of the water in the water flow sandwich can be accurately adjusted to reach and maintain the temperature range required for the experiment. For example, before the experiment begins, the power and working time of the heating rod are set according to the experimental requirements, and the water in the water flow sandwich is heated to the predetermined test temperature. During the experiment, the heating rod can automatically adjust the power or work intermittently according to the water temperature information fed back by the temperature sensor to keep the water temperature constant. If the water temperature is lower than the set temperature, the heating rod will increase the power output or continue to work; conversely, if the water temperature is too high, the heating rod will reduce the power or stop working to ensure that the water temperature is always stable within a range suitable for aquatic biological metabolism and excretion determination.

[0030] It should be noted that the presence of the heating chamber 12 provides a stable temperature environment for the intermediate layer metabolic measurement module. During the measurement of the metabolic rate and excretion rate of aquatic organisms, the stable temperature helps to maintain the normal physiological activities of aquatic organisms and reduce the stress response caused by temperature changes, so that the measured metabolic rate and excretion rate can more truly reflect the physiological indicators of organisms in a natural state.

[0031] It should also be noted that the box 8 is designed with square grooves (gasket placement track 7) around the metabolic measurement module for installing the rubber sealing ring. The design of the square groove (gasket placement track 7) provides a precise installation position for the rubber sealing ring. After the rubber sealing ring is installed in the groove, it can form a tight sealing interface between the box 8 and the metabolic measurement module. When the device is measuring the metabolic rate and excretion rate of aquatic organisms, this sealing structure can effectively prevent external air, impurities, etc. from entering the device, while also preventing water and gas leakage in the device, ensuring the stability and sealing of the measurement environment. For example, when measuring the metabolic rate, it is necessary to keep the water environment in the device relatively stable. A well-sealed device can prevent the accidental exchange of gases such as oxygen, ensure the accuracy of measuring the change in dissolved oxygen concentration, and thus improve the reliability of metabolic rate calculation.

[0032] In addition, good sealing performance can also maintain the constant water temperature in the heating chamber 12, because the heat will not be lost due to leakage, which helps to provide stable temperature conditions for aquatic organisms and further ensure the accuracy of experimental results.

[0033] Please continue to refer to Figure 1The device also includes a top cover plate 3, and three cylindrical channels are arranged at different positions above the top cover plate 3, namely a dissolved oxygen meter probe access hole 1, a water inlet 2 and a water outlet 17; the dissolved oxygen meter probe access hole 1 is used to measure the dissolved oxygen concentration in the water body in the device, the water inlet 2 is used to pass external water into the measuring device, and the water outlet 17 is used to discharge excess water after the water fills the measuring device.

[0034] It should be noted that the sizes of the three cylindrical channels arranged at different positions above the top cover plate 3 can be set according to actual needs and are not limited here. The dissolved oxygen meter probe access hole 1 provides a channel for the dissolved oxygen meter probe to enter the water body inside the device. During the experiment, the dissolved oxygen meter probe is inserted into the water body through the hole, and the dissolved oxygen concentration in the water body can be measured in real time and accurately. By monitoring the changes in dissolved oxygen concentration at different time points, the oxygen consumption rate of the organism can be accurately calculated, thereby evaluating its metabolic level. The water inlet 2 is used to pass external water into the measuring device. Through an external water pump or other water supply equipment, the treated water (such as specific temperature, water quality, etc.) can be slowly injected into the device. The main function of the water outlet 17 is to discharge excess water after the water fills the measuring device to ensure that the water level in the device is in a suitable state. In addition, a peristaltic pump is connected to the outside of the water inlet, and a water outlet pipe is connected to the outside of the water outlet. The water outlet pipe is in a closed state during the measurement process. When the dissolved oxygen level in the middle layer metabolic module is found to be too low through the dissolved oxygen meter probe, for example, generally below 50% saturated dissolved oxygen, in order to reduce the stress behavior of the organism caused by hypoxia, the external oxygen-rich water can be pumped into the interior through a peristaltic pump, and the internal oxygen-deficient water can be discharged through the outlet pipe, which is suitable for long-term measurement of the metabolic level of the organism.

[0035] Figure 5 This is a ventral view of the top cover shown in this application, Figure 6 This is a front view of the top cover shown in this application, please also refer to Figure 1 , Figure 5 and Figure 6 , spiral holes 4 are provided at the four corners of the top cover plate 3, and spiral holes 4 corresponding to the top cover plate 3 are provided at the four corners of the box body 8. The cross-sectional sizes of the top cover plate 3 and the box body 8 are the same, and the top cover plate 3 contacts the box body 8 through the spiral holes 4 to close the box body 8. The top cover plate 3 can be tightly fixed to the box body 8 by passing the spiral holes 4 through bolts and other connecting parts. Since the spiral holes 4 at the four corners of the box body 8 correspond to the spiral holes 4 of the top cover plate 3, the accuracy and tightness of the connection are ensured. In the process of tightening the bolts, the pressure between the top cover plate 3 and the box body 8 gradually increases, so that the sealing performance between the two is enhanced, so that a relatively closed environment is formed inside the device, preventing external factors (such as air, impurities, etc.) from entering the device to interfere with the experiment, and also preventing water and gas leakage in the device.

[0036] For metabolic rate determination, a sealed environment helps maintain the stability of dissolved oxygen concentration in the water body in the device, ensuring that the dissolved oxygen meter probe can accurately measure changes in dissolved oxygen, thereby accurately calculating the oxygen consumption rate of aquatic organisms. If the device is not well sealed, outside air can easily enter, resulting in inaccurate dissolved oxygen concentration measurements, which in turn affects the reliability of metabolic rate calculations. For excretion rate determination, a sealed device can prevent external substances from contaminating the water samples in the pumping tank, ensuring that the extracted water samples can truly reflect the excretion of aquatic organisms and improve the accuracy of excretion rate calculations.

[0037] In addition, the cross-sectional size of the top cover plate 3 is the same as that of the box body 8, which can enhance the structural stability of the device. During the experiment, water circulation, biological activities, etc. may occur inside the device, and these factors may generate certain forces. When the top cover plate 3 and the box body 8 are matched in size and tightly connected, the entire device can better withstand these forces and reduce the relative displacement and shaking between components. The stable structure helps to ensure the normal operation of various measuring equipment (such as dissolved oxygen meter probe, circulating water pump, etc.) during the experiment, reduce experimental errors caused by device shaking or loose components, and improve the reliability and repeatability of experimental results.

[0038] The present application provides a device for synchronously measuring the metabolic rate and excretion rate of aquatic organisms. On the first hand, a metabolic chamber and a pumping chamber are set in the middle layer metabolic measurement module, and are equipped with corresponding measurement and circulation equipment, as well as a dissolved oxygen meter probe access hole on the top cover plate for measuring the dissolved oxygen concentration to calculate the metabolic rate, and the relevant structure of the pumping chamber is used to extract water samples to measure the excretion rate, thereby avoiding the long experimental cycle and error problems caused by changes in biological states brought about by traditional step-by-step measurement methods, improving the experimental efficiency, making the obtained metabolic rate and excretion rate data more timely and relevant, and being able to more comprehensively and accurately reflect the physiological state of aquatic organisms.

[0039] Secondly, a sealed heating chamber is formed by the water flow interlayer between the box and the metabolic measurement module, and a heating rod placement groove and a heating rod are provided in the heating chamber, which can accurately adjust the water temperature and keep it constant according to the experimental requirements, reduce the stress response of aquatic organisms caused by temperature changes, ensure that the metabolism and excretion processes are carried out under stable conditions, and improve the accuracy of the measurement results. In addition, by setting the water pumping port and the rubber ball binding hole, the partial pressure inside and outside the water body can be kept consistent when extracting water samples, ensuring that the pumping operation is smooth and does not affect the environment inside the device. At the same time, the box and the top cover are tightly connected through spiral holes, and a square groove is designed at the connection to install a rubber sealing ring to ensure the overall sealing of the device. Good sealing performance prevents external air, impurities, etc. from entering the device to interfere with the experiment, as well as water and gas leakage in the device, maintaining the stability of the experimental environment, and plays a key role in the accurate measurement of metabolic rate (such as dissolved oxygen concentration determination) and excretion rate (such as water sample composition determination).

[0040] On the third aspect, by setting up an aquatic organism limiting device and utilizing the combination of the card slot, the breathing bottle and the second partition plate, it is possible to flexibly adjust according to different specifications of aquatic organisms, accurately limit biological activities, keep them in a resting state, reduce metabolic differences between individuals, and improve the accuracy of metabolic rate determination. In addition, the present application also takes into account the problem of different bacterial oxygen consumption caused by each change of water body by setting up a bacterial oxygen consumption bin, which can improve the accuracy of metabolic rate calculation, and the bacterial oxygen consumption bin can monitor the bacterial oxygen consumption in real time through the dissolved oxygen meter probe, and accurately measure the bacterial oxygen consumption and deduct it when calculating the oxygen consumption rate of aquatic organisms, so that the experimental data is more real and accurate.

[0041] Figure 7 Flow chart of the method for simultaneous determination of aquatic organism metabolic rate and excretion rate provided in this application. Please refer to Figure 7 , based on the above device embodiment, the method includes: S701. After water is injected into the intermediate layer metabolic determination module, an aquatic organism limiting device is placed in the metabolic chamber, and the aquatic organism is placed in the breathing bottle.

[0042] It should be noted that after water is injected into the intermediate layer metabolic determination module, an aquatic organism limiting device is placed in the metabolic chamber, and the aquatic organism is placed in the breathing bottle, including: (1) Fill the intermediate layer metabolic measurement module with water for the first time, so that the water level remains below the heating rod placement groove.

[0043] It should be noted that a lower water level can prevent water from overflowing or causing unnecessary water flow disturbance when placing the aquatic organism limit device. Secondly, the water level at this time provides a certain operating space for the subsequent installation and adjustment of the equipment, making it convenient to accurately place the aquatic organism limit device in the appropriate position in the metabolic chamber. At the same time, the reserved space is also convenient for checking and ensuring that the internal components of the device (such as the circulation pump, partition plate, etc.) are in normal working condition, preparing for the smooth progress of subsequent experiments.

[0044] (2) placing an aquatic organism limiting device in the metabolic chamber, and injecting water into the intermediate layer metabolic measurement module for a second time, and placing the aquatic organism in the breathing bottle after the water level covers the intermediate layer metabolic measurement module.

[0045] It should be noted that the second water injection is carried out after the aquatic organism limiting device is placed in the metabolic chamber, and the aquatic organisms are placed in the breathing bottle after the water level covers the middle layer metabolic measurement module. This ensures that when the aquatic organisms enter the measurement environment, the surrounding water environment is basically stable. Placing the limiting device first can prevent its position from being shifted or damaged during the water injection process, and placing the aquatic organisms at the appropriate water level can enable them to adapt to the relatively stable water environment as soon as possible and reduce the stress response caused by environmental changes. The second water injection covers the entire module, ensuring that the aquatic organisms are completely in a suitable water environment, and at this time, equipment such as the circulating pump can work normally in a water-filled environment, promoting water circulation and material exchange, and providing good conditions for accurately measuring metabolic rate and excretion rate.

[0046] S702. After the oxygen exposure operation is performed and when the dissolved oxygen concentration reaches a preset threshold, the box is sealed using a top cover.

[0047] It should be noted that the purpose of aeration is to rapidly increase the dissolved oxygen concentration in the water in the device to a relatively high level before the experiment begins. Sufficient initial dissolved oxygen can ensure that there is enough oxygen for aquatic organisms to consume during the experiment, and can also make the dissolved oxygen concentration have a significant gradient during the biological consumption process, making it easier to accurately measure its changes through the dissolved oxygen meter probe, thereby accurately calculating the oxygen consumption rate. Among them, the preset threshold is generally above 8.5 mg / mL.

[0048] When the dissolved oxygen concentration reaches the preset threshold, the top cover is used to seal the box in order to establish a relatively closed experimental environment. In a sealed environment, the consumption of dissolved oxygen in the water is mainly caused by the respiration of aquatic organisms. This allows the dissolved oxygen concentration that changes over time to be more accurately measured by the dissolved oxygen meter probe, and then the oxygen consumption rate of the organisms can be calculated. At the same time, a sealed environment also helps to maintain the stability of other experimental conditions, such as temperature and pressure, reduce the impact of external factors on the experimental results, and provide a reliable environmental basis for the simultaneous determination of metabolic rate and excretion rate.

[0049] S703. After a specified time, the oxygen consumption rate is measured by the dissolved oxygen meter probe, and water is drawn through the water pumping port to calculate the excretion rate.

[0050] Specifically, the dissolved oxygen meter probe continuously monitors the dissolved oxygen concentration in the water in the device. Since the respiration of aquatic organisms in a sealed environment consumes oxygen, the dissolved oxygen concentration will gradually decrease over time. Based on the dissolved oxygen concentration data at different time points measured by the dissolved oxygen meter probe, combined with the initial dissolved oxygen concentration at the beginning of the experiment, the water volume, and the duration of the experiment, the corresponding calculation formula (such as a specific metabolic rate calculation model) can be used to accurately calculate the oxygen consumption rate of aquatic organisms during this period, thereby evaluating their metabolic level.

[0051] Water is extracted through the water pumping port, and then the ammonia nitrogen concentration in the extracted water sample is determined by a suitable chemical analysis method. The initial ammonia nitrogen concentration at the beginning of the experiment can be measured by placing the aquatic organisms in the limiting device and sealing the device for half an hour, then extracting the water sample in the device for ammonia nitrogen determination (aquatic organisms will produce a stress response when entering a new environment, and the data measured after a period of rest will be more accurate). After the specified time, water samples are extracted again to determine the final ammonia nitrogen concentration. According to factors such as the change in ammonia nitrogen concentration, water volume, and experimental time, the excretion rate is calculated using relevant formulas to understand the ability of aquatic organisms to excrete ammonia metabolites. It should be noted that during the water sampling process, the rubber ball connected to the cylindrical channel in the water pumping tank ensures that the partial pressure inside and outside the water body is consistent, ensuring that the extracted water sample is representative and improving the accuracy of the excretion rate calculation.

[0052] The method for synchronously measuring the metabolic rate and excretion rate of aquatic organisms provided in the present application can realize the synchronous measurement of the metabolic rate and excretion rate of aquatic organisms, and improve the accuracy and reliability of the measurement.

[0053] The method of this embodiment can be used to execute Figure 1 The steps, specific implementation principles and implementation processes of the device embodiment shown are similar and will not be repeated here.

[0054] The above description is only a preferred embodiment of the present application and is not intended to limit the present application. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present application shall be included in the scope of protection of the present application.

Claims

1. A device for synchronously measuring the metabolic rate and excretion rate of aquatic organisms, characterized in that: The device comprises a box body, in which an intermediate layer metabolism determination module is arranged; The intermediate layer metabolic measurement module comprises a metabolic chamber and a water pumping chamber which are arranged adjacent to each other, and the metabolic chamber and the water pumping chamber are separated by a first partition plate; An aquatic organism limiting device is provided at the upper part of the metabolic chamber, and the aquatic organism limiting device is communicated with the water flow inside the metabolic chamber; The bottom of the aquatic organism limiting device is provided with a plurality of slots for adjusting in the horizontal direction, and a breathing bottle is arranged above the slots; a plurality of second partition plates are arranged at intervals in the slots, and the height of the second partition plates is higher than that of the breathing bottle, and the side surface of each second partition plate is in contact with the breathing bottle; the size of the space separated by the breathing bottle and the corresponding second partition plates is the same as the size of the aquatic organism to be tested, and the separated space is used to place the aquatic organism to be tested; a plurality of hollow holes are arranged on the breathing bottle and the second partition plates, and the separated space is connected with the outside of the aquatic organism limiting device through the plurality of hollow holes.

2. The device according to claim 1, characterized in that A circulation pump is provided on the side of the first partition plate, and the circulation pump is located below the aquatic organism limiting device. The circulation pump drives the water flow inside the intermediate layer metabolism determination module to circulate.

3. The device according to claim 1, characterized in that The device further comprises two cylindrical channels arranged in parallel, wherein a first port of the cylindrical channel is arranged inside the water pumping bin, and a second port of the cylindrical channel is arranged outside the box body, for connecting the inside of the water pumping bin with the outside of the box body; Among them, one second port is connected to the pumping device, and the other second port is connected to the rubber ball.

4. The device according to claim 1, characterized in that There is a water flow interlayer between the box body and the metabolic determination module, and the water flow interlayer is sealed to form a heating chamber; The heating chamber is provided with a heating rod placement groove on one side close to the outside of the box body. A heating rod is arranged in the heating rod placement groove. The heating rod heats the water in the water flow interlayer.

5. The device according to claim 1, characterized in that The intermediate layer metabolism measurement module is also provided with a bacterial oxygen consumption bin on the pumping bin side. The bacterial oxygen consumption bin is sealed and has a water inlet and outlet and a dissolved oxygen meter probe access hole. The bacterial oxygen consumption bin is used to measure the total oxygen consumption of bacteria in the water body during the metabolic process.

6. The device according to claim 1, characterized in that The box body is designed with square grooves around the periphery of the metabolic determination module for installing a rubber sealing ring.

7. The device according to claim 1, characterized in that The device also includes a top cover plate, and three cylindrical channels are arranged at different positions above the top cover plate, which are respectively a dissolved oxygen meter probe access hole, a water inlet and a water outlet; The dissolved oxygen meter probe access hole is used to measure the dissolved oxygen concentration in the water body of the metabolic bin in the device, the water inlet is used to pass external water into the measuring device, and the water outlet is used to discharge excess water after the measuring device is filled with water.

8. The device according to claim 7, characterized in that The top cover plate has four corners with spiral holes, the box body has four corners with spiral holes corresponding to the top cover plate, the top cover plate has the same cross-sectional size as the box body, and the top cover plate contacts the box body through the spiral holes to close the box body.

9. A method for synchronously measuring the metabolic rate and excretion rate of aquatic organisms, characterized in that: The synchronous determination method is applied to the device according to any one of claims 1 to 8, and the method comprises: After water is injected into the intermediate layer metabolism determination module, an aquatic organism limiting device is placed in the metabolism chamber, and the aquatic organism is placed in the breathing bottle; After the oxygen exposure operation, and when the dissolved oxygen concentration reaches the preset threshold, the box is sealed with the top cover; During a specified period of time, the oxygen consumption rate is measured by the dissolved oxygen meter probe, and water is pumped through the water extraction port to calculate the excretion rate.

10. The method according to claim 9, characterized in that After water is injected into the intermediate layer metabolism determination module, an aquatic organism limiting device is placed in the metabolism chamber, and the aquatic organism is placed in the breathing bottle, including: Filling water into the intermediate layer metabolism measurement module for the first time, so that the water level stays below the heating rod placement groove; An aquatic organism limiting device is placed in the metabolic chamber, and water is injected into the intermediate layer metabolic measurement module for a second time. After the water level covers the intermediate layer metabolic measurement module, the aquatic organism is placed in the breathing bottle.

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