CN106442959B - Daphnia magna continuous-flow type toxicity test device - Google Patents

Abstract

The invention relates to a large daphnia flowing water toxicity test device, which comprises a solution preparation system, a solution temporary storage system, a toxicity exposure system, a solution automatic circulation and supply system, and a waste liquid treatment system; the solution preparation system prepares a specific concentration for the daphnia magna toxicity test The test solution; the solution temporary storage system provides conditions for the homogenization and temporary storage of the test solution; the toxicity exposure system provides suitable space, temperature and light for the exposure of Daphnia magna in the test solution; the automatic circulation and replenishment system of the solution Connected between the solution preparation system and the temporary storage system, between the solution temporary storage system and the toxicity exposure system, it can realize the automatic supply of the solution from the preparation system to the temporary storage system, and the delivery of the solution from the solution temporary storage system to the toxicity test system ; The waste liquid treatment system provides purification functions such as filtration and adsorption for the waste liquid generated by the toxic exposure system.

Description

Large Daphnia Flowing Water Toxicity Test Device

technical field

The invention belongs to the field of ecotoxicology experiments, and in particular relates to a large daphnia running water toxicity test device.

Background technique

Daphnia magna Straus belongs to the phylum Arthropoda, Crustacea, and Cladocera. It is a common planktonic crustacean and an indispensable component in the freshwater food chain. Because of its short life cycle, fast growth, high reproductive capacity, easy cultivation, and sensitivity to water environment stress, it has become a model organism widely used in laboratories at home and abroad. important role in testing.

At present, many international organizations and countries such as OECD, European Union, the United States, Japan, South Korea and my country have formed systematic standard methods for toxicological testing of Daphnia magna. In order to ensure the scientificity, validity and comparability of the test results, most of these methods require to confirm whether the test substance can remain relatively stable in the test system throughout the test cycle (that is, the measured concentration is not lower than the initial concentration or the prepared concentration). 80%). If the test substance is relatively stable, the general static method (the solution is not renewed during the entire test period) can be used for the toxicity exposure test; otherwise, the semi-static method (the solution is regularly renewed) or the flow-through test method must be used. Since the maximum solution update frequency of the semi-static method can generally only reach 24 hours, for unstable test substances that are easily hydrolyzed, volatile, and photolyzed, if the degradation half-life is less than 24 hours, flow-through test methods must be used. This unstable test substance belongs to the "difficult substances" (difficult substances), which accounts for a considerable part of industrial chemicals and pesticide intermediates, and is the main difficulty in water ecotoxicity testing and an important factor affecting the accuracy of testing.

In addition, chronic toxicity such as reproduction and development, and environmental endocrine disruptive toxicity are generally longer and must cover most or the entire life cycle of organisms. In order to reduce the deviation caused by the degradation or escape of the test substance, the aquatic ecotoxicity test of the chronic toxicity of chemical substances and environmental endocrine disruption must also ensure that the test solution is at a relatively stable concentration within a long toxicity test period. Expose the test organism. Therefore, the flow-through test method is also an important technology to solve the key problems of the chronic toxicity of chemical substances and environmental endocrine disruption research tests.

The so-called flow-through test method is to keep the test solution automatically updated, and the solution will not enter the test system after being discarded. This method requires a huge amount of work and requires the help of relevant automated instruments and equipment. As for the large-scale daphnia running water toxicity test, the corresponding automation equipment must not only automatically and accurately prepare the test solution with a specific concentration, automatically update the test solution, and consider the treatment of the terminal waste liquid, but also need to meet the requirements of the small daphnia magna. Survival and reproduction conditions of Cladocera and necessary experimental operation requirements such as observation, counting, and larvae collection. Due to the complex, delicate and special technical requirements mentioned above, there is no equipment or device that meets the requirements on the market at present. The laboratories that have carried out large-scale daphnia toxicity research in my country can only use static and semi-static methods to test manually. This method is not only inefficient, but also difficult to meet the testing requirements of unstable "difficult substances" and the long-term research requirements of chronic toxicity of chemical substances and environmental endocrine disrupting aquatic ecotoxicity. This brings great difficulties and errors to chemical substance testing and scientific research.

Contents of the invention

In order to facilitate the smooth progress of the large daphnia running water toxicity test, the invention provides a large daphnia running water toxicity test device.

The test device includes a solution preparation system, a solution temporary storage system, a toxicity exposure system, a solution automatic circulation and supply system, and a waste liquid treatment system; the solution preparation system and the solution temporary storage system, the solution temporary storage system and the toxicity exposure system The rooms are connected to the supply system through the automatic circulation of the solution, and the toxic exposure system is connected to the waste liquid treatment system;

The solution preparation system includes 1 set of automatic liquid dispenser, 1 medium storage barrel, 1 level I mother solution bottle, 1 level II mother solution bottle, 1 co-solvent bottle, 1 co-solvent control preparation bottle, 1 The blank control preparation bottle, the 5 test solution preparation bottles, the second grade mother liquor bottle, the co-solvent control preparation bottle, the blank control preparation bottle, and the test solution preparation bottle were placed on independent magnetic stirrers;

The automatic liquid dispenser includes 3 dual-channel peristaltic pumps, 2 single-channel peristaltic pumps, 1 liquid level sensor, 1 screw transmission mechanism, 1 eight-way valve, and a PLC controller; It is connected with the mouth of the second level mother liquid bottle, the bottom of the second level mother liquid bottle and the first test solution preparation bottle through the two infusion tubes of the same dual-channel peristaltic pump; the bottom of the first test solution preparation bottle is connected with The bottle mouth of the second test solution preparation bottle, the bottom of the second test solution preparation bottle are connected with the bottle mouth of the third test solution preparation bottle by two infusion tubes of the same dual-path peristaltic pump; The bottom of the first test solution preparation bottle and the mouth of the fourth test solution preparation bottle, the bottom of the fourth test solution preparation bottle and the fifth test solution preparation bottle pass through the same dual-channel peristaltic pump The bottom of the medium storage tank is connected to the second level mother liquor bottle, the cosolvent control preparation bottle, the blank control preparation bottle, and the five test samples through the infusion pipe of the single-channel peristaltic pump, the screw transmission mechanism and the eight-way valve Solution preparation bottle; the liquid level sensor is placed above the level II mother liquor bottle;

The solution temporary storage system includes 1 cosolvent control temporary storage bottle, 1 blank control temporary storage bottle, and 5 test solution temporary storage bottles. The outlets of the control preparation bottle and the 5 solution preparation bottles are connected through pipelines, and solenoid valves are provided on the pipelines; all temporary storage bottles are made of brown glass, and are provided with top covers, and the top covers are provided with holes for the infusion tubes of peristaltic pumps to be inserted into; All reagent bottles are placed on independent magnetic stirrers, and the solution temporary storage system is placed under the solution configuration system;

The solution automatic circulation and supply system includes 2 liquid level sensors, 1 automatic solenoid valve, 1 seven-channel peristaltic pump, 1 PLC controller, and a liquid level sensor installed in the fifth test solution preparation bottle, Another liquid level sensor is installed in the fifth solution temporary storage bottle; the cosolvent control preparation bottle, the blank control preparation bottle and the 5 solution preparation bottles respectively pass the seven infusion tubes of the seven-channel peristaltic pump and the cosolvent control temporary storage bottle, blank control temporary storage bottle, and 5 test solution temporary storage bottles are connected independently, and the solenoid valve is installed in the middle of the seven infusion tubes, and the water flow is automatically blocked or connected by the PLC controller;

The toxicity exposure system includes 1 constant temperature water bath, 1 group of light with automatic photoperiod control, 1 cosolvent control test tank, 1 blank control test tank, 5 test solution test tanks, and the above 7 test tanks are placed In the constant temperature water bath, the entrances of the 7 test cylinders are connected to the above-mentioned 7 corresponding temporary storage bottles through seven independent infusion tubes of the seven-way peristaltic pump;

The waste liquid treatment system includes a layer of filter cotton and a layer of activated carbon filter column, both of which are integrated in a closed system, and are only connected to the outlet holes of each test cylinder of the toxicity exposure system through the water inlet pipe.

Specifically, there are universal wheels at the bottom of the medium storage barrel in the solution preparation system, and all reagent bottles are made of brown glass; the medium storage barrel and reagent bottles have top covers, and the top covers are provided with holes for the peristaltic pump The infusion tube is inserted.

Specifically, the test cylinders are all made of transparent glass, equipped with a top cover, and the cylinder wall is provided with water inlet holes and water outlet holes. In each test cylinder, 10 small shrouds and a detachable shroud frame are placed. Made of stainless steel mesh, the aperture is 60 mesh to 80 mesh, and the upper end is open; the shroud frame is a glass plate with holes, the number of holes is 10, and the aperture is the same as the outer diameter of the shroud; the height of the shroud frame is slightly lower than the test cylinder mouth. 10 shrouds are inserted into the test cylinder from the opening of the shroud frame, and the bottom of the shroud is slightly higher than the bottom of the tank, and the mouth of the shroud is slightly higher than the shroud frame; the water inlet and outlet holes are located under the shroud frame, and the water inlet hole Close to the bottom of the test cylinder, the water outlet hole is close to the shroud frame, and the water inlet hole and the water outlet hole are respectively located at the two ends of the cylinder wall opposite to the test cylinder and with the longest straight line distance.

Concrete, the control method of described PLC controller is as follows:

When the "start" button on the PLC controller is activated or the liquid level sensor above the level II mother liquid bottle sends out a signal lower than the warning value, the pump will absorb the solution from the level I mother liquid bottle and add it to the second level mother liquid bottle; The outlet of the eight-way valve is aligned with the second-level mother solution bottle, and the M7 medium is drawn from the medium storage barrel and added to the second-level mother solution tank; after 20 minutes, the solution is drawn from the second-level mother solution bottle and added to the first test solution preparation bottle , the outlet of the eight-way valve is aligned with the first test solution preparation bottle, draw the M7 medium from the medium storage barrel and add it to the first test solution preparation bottle; after 20 minutes, draw from the first test solution preparation bottle solution and added to the second test solution preparation bottle, then draw the M7 medium from the medium storage barrel and add the second test solution preparation bottle; after 20min, draw the solution from the second test solution preparation bottle Add it to the third test solution preparation bottle, then draw the M7 medium from the medium storage barrel and add it to the third test solution preparation bottle; after 20 minutes, draw the solution from the third test solution preparation bottle and add it to the third test solution preparation bottle After 20 minutes, draw the solution from the fourth test solution preparation bottle and add it to the fifth test solution preparation bottle. Then draw the M7 medium from the medium storage barrel and add it to the fifth test solution preparation bottle; then draw the solvent from the solvent bottle and add it to the solvent control preparation bottle, and then draw the medium from the medium storage barrel Aspirate the M7 medium from the medium and add it to the solvent control preparation bottle; draw the M7 medium from the medium storage barrel and add it to the blank control preparation bottle;

When the liquid level sensor above the fifth test solution temporary storage bottle sends a signal that the liquid level is lower than the warning value, the program will open the solenoid valve, and the 5 solution preparation bottles, blank control preparation bottles, and solvent control preparation bottles rely on gravity Enter 5 solution temporary storage bottles, blank control temporary storage bottles, and solvent control temporary storage bottles respectively; when the liquid level sensor above the fifth solution preparation bottle sends out a signal that the liquid level is lower than the warning value, the program will close the solenoid valve, The liquid in the preparation tank and the temporary storage tank is blocked; then draw the solution from the second-level mother liquid bottle and add it to the first test solution preparation bottle, and use the screw drive mechanism to align the outlet of the eight-way valve with the first test solution preparation bottle, draw the M7 medium from the medium storage barrel and add it to the first test solution preparation bottle; after 20 minutes, draw the solution from the first test solution preparation bottle and add it to the second test solution preparation bottle, and then Draw the M7 medium from the medium storage barrel and add it to the second test solution preparation bottle; after 20 minutes, draw the solution from the second test solution preparation bottle and add it to the third test solution preparation bottle, and then from the culture medium Draw the M7 medium from the base storage barrel and add it to the third test solution preparation bottle; after 20 minutes, draw the solution from the third test solution preparation bottle and add it to the fourth test solution preparation bottle, and then store it from the medium Draw the M7 medium from the barrel and add it to the fourth test solution preparation bottle; after 20 minutes, draw the solution from the fourth test solution preparation bottle and add it to the fifth test solution preparation bottle, and then add it to the medium storage barrel Aspirate the M7 medium and add it to the fifth test solution preparation bottle; then aspirate the solvent from the cosolvent bottle and add it to the cosolvent control preparation bottle, then aspirate the M7 medium from the medium storage bucket and add it to the cosolvent control preparation bottle medium; draw M7 medium from the medium storage barrel and add it to the blank control preparation bottle;

The solution is continuously pumped from the solution temporary storage bottle into the test cylinder through the seven-channel peristaltic pump, and the test solution exceeding the capacity of the test cylinder is continuously discharged from the water outlet to the waste liquid treatment system.

The beneficial effects of this device are:

1. Realize the automatic preparation of the test solution with a specific concentration;

2. Realize the flow-type automatic update and replenishment of the test solution;

3. The closed design of the test tank can reduce the volatilization loss of the test solution, and also plays an important role in maintaining the stability of the water temperature in the test tank; the small stainless steel enclosure is made of inert materials to avoid the chemical effect of the test material on most test drugs and interfere with it. Toxicity; the design of the small shroud and the shroud stand is convenient for users to take and place, and at the same time, it can fully contact the large daphnia with the test solution under flowing water conditions and complete life processes such as development and reproduction, and at the same time avoid female daphnia and juvenile daphnia. escape and mechanical damage. And the bottom and surrounding of the shroud are not in contact with the wall of the test tank, so as to keep the water temperature difference between the biological activity areas inside the shroud as low as possible, and not affected by the fluctuation of the water temperature outside the test tank; the design of the whole test tank is convenient for the user to adjust each The reproduction and death of daphnia in the small enclosures are observed and counted, and the marked enclosures can prevent users from repeatedly counting different small enclosures or confusing observation results; after the test is completed, the detachable design of the entire test cylinder is also convenient for cleaning.

4. Realize the full contact of Daphnia large daphnia with the test solution under flowing water conditions and complete life processes such as development and reproduction, while avoiding the escape and mechanical damage of mother daphnia and juvenile daphnia.

5. Realize the treatment of test wastewater such as filtration and adsorption to reduce environmental hazards.

6. The experimental device is convenient for experimental operation, low in cost, and easy to popularize in the laboratory.

Description of drawings

Figure 1 External view of the overall assembly diagram.

Figure 2 Internal view of the overall assembly. in:

Figure 3 Detail view of the test cylinder. in:

Fig. 4 Detailed view of waste liquid treatment system.

Among them: 1. Culture medium storage barrel, 2. Solution preparation room, 3. Solution temporary storage room, 4. Toxicity exposure room, 5. PLC controller, 6. Transmission machinery room, 7. Waste liquid treatment room, 80,000 Direction wheel, 9. handle, 10. solution preparation bottle, 11. solution temporary storage bottle, 12. magnetic stirrer, 13. constant temperature water bath, 14. test cylinder, 15. photoperiod automatic control of light 16. peristaltic pump, 17. Waste liquid treatment system, 18. Stainless steel mesh small apron, 19. Water inlet, 20. Water outlet, 21. Dome cover, 22. Apron frame, 23. Apron socket, 24. Water inlet, 25. Upper water storage layer, 26. filter cotton, 27. activated carbon filter column, 28. water outlet.

Detailed ways:

The present invention will be described in further detail below in conjunction with the accompanying drawings.

Example 1

As shown in Figure 1, the large daphnia running water toxicity test device, in which the symbols or letters are represented as follows: 1 is the medium storage barrel, used to store the daphnia magna culture medium; 2 is the solution preparation room for the solution preparation system, with different concentrations The test solution is prepared here. The solution preparation system includes reagent bottles (level I mother solution bottle, level II mother solution bottle, solvent bottle, cosolvent control preparation bottle, blank control preparation bottle, test solution preparation bottle), magnetic stirrer, Peristaltic pump, PLC controller; 3 is the solution temporary storage room for the solution temporary storage system, where the prepared test solution is temporarily stored and fully homogenized. The solution temporary storage system includes reagent bottles (cosolvent control temporary storage bottle , blank control temporary storage bottle, test solution temporary storage bottle) and magnetic stirrer; 4 is the toxic exposure room for the toxic exposure system; 5 is the PLC controller; 6 is used for placing the peristaltic pump, screw drive mechanism, solenoid valve 7 is the waste liquid treatment room for placing the waste liquid treatment system; 8 is the universal wheel to facilitate the movement of the whole set of devices; 9 is the handle of the small door outside each sub-chamber; 10 is the solution configuration system Solution preparation bottle, used to prepare solution; 11 is the solution temporary storage bottle of the solution temporary storage system, used for solution temporary storage and homogenization; 12 is a magnetic stirrer, used for solution stirring during solution preparation and homogenization ; 13 is the constant temperature water bath of the toxicity test system, used to maintain the temperature of the test system; 14 is the test tank of the toxicity test system, used for the toxicity exposure of Daphnia magna; 15 is the light that can automatically control the photoperiod in the toxicity test system; 16 is the solution preparation system, the peristaltic pump and other equipment required in the solution automatic circulation and replenishment system; 17 is the waste liquid treatment system;

Figure 2 is a detailed view of the test tank in the toxicity exposure system, in which the symbols or letters are indicated as follows: 18 is a small stainless steel casing, which is a device for Daphnia magna to accept drug exposure and prevent it from escaping; 19 is a water inlet, used for uninterrupted Input fresh test solution; 20 is the water outlet, used to discharge the overflowed test solution; 21 is the dome cover, used to reduce the volatilization loss of the test solution and reduce the drug contact of the test personnel; 22 is the shroud frame, used to support the stainless steel shroud ; 23 is the jack on the shroud frame, which is used to insert the small shroud; R1~R10 is the numbering of 10 shroud jacks, which can be inserted into 10 small shrouds in turn, as 10 parallels of the same concentration treatment group;

Fig. 3 is a detailed diagram of the waste liquid treatment system, wherein each symbol or letter represents as follows: 24 is a water inlet, is used for collecting the experimental waste liquid that produces from the test cylinder of toxicity exposure system; 25 is the upper water storage layer, is used for storing The experimental waste liquid discharged from the test cylinder; 26 is a filter cotton, used for primary filtration of large particles in the waste liquid; 27 is an activated carbon filter column, used for adsorption and purification of drugs in the waste liquid; 28 is a water outlet, used for Discharge the treated effluent.

The names and sizes of the main components of the device are as follows:

Culture medium storage barrel 1: made of stainless steel, with a diameter of 60 cm, a height of 85 cm, and a volume of 120 L;

Solution preparation chamber 2: made of stainless steel, length 80cm, width 70cm, height 50cm;

Solution temporary storage room 3: stainless steel material, length 60cm, width 70cm, height 50cm;

Toxic exposure chamber 4: length 100cm, width 70cm, height 60cm;

PLC controller 5: length 20cm, width 70cm, height 60cm;

Transmission machine room 6: made of stainless steel, length 40cm, width 70cm, height 60cm;

Solution preparation bottle 10 and solution temporary storage bottle 11 (Class I mother liquor bottle, II mother liquor bottle, solvent bottle, cosolvent control preparation bottle, blank control preparation bottle, test solution preparation bottle and cosolvent contrast temporary storage bottle, blank control Temporary storage bottle, test solution temporary storage bottle): brown proportional material, the opening of the top cover can be inserted into the infusion tube, the volume is 1L, the diameter is 10cm, and the height is 23cm;

Magnetic stirrer 12: length 12cm, width 15cm, height 7cm;

Water bath 13: stainless steel material, length 100cm, width 70cm, height 12cm;

Test cylinder 14: made of glass, with a diameter of 23cm, a height of 15cm, and a volume of 4L;

Light 15: cold light source, can automatically control the photoperiod, light intensity 2000±300lux;

Peristaltic pump 16: pump speed range 0.02~200 mL/min, all equipped with RS485 communication interface and support MOUDBUS protocol;

Waste liquid treatment system 17: stainless steel shell, length 55cm, width 65cm, height 45cm;

Small apron 18: stainless steel mesh, diameter 50mm; mesh 60 mesh; height 70mm;

Dome cover 21: made of glass, with a diameter of 23cm and a top height of 6cm;

Apron frame 22: made of glass, with a diameter of 23cm;

Shroud socket 23: a round hole with a diameter of 51 mm;

Waste liquid treatment system water inlet 24: internal diameter 3cm;

Upper water storage layer 25: length 65cm, width 70cm, height 20cm;

Filter cotton 26; synthetic fiber material;

Activated carbon filter column 27: activated carbon filter column, diameter 3cm, height 20cm;

Water outlet 25 of the waste liquid treatment system: the inner diameter is 3 cm.

Example 2

Assuming that a large-scale daphnia running water toxicity test is required, the test concentration is designed as: blank control, cosolvent control, 0.1mg/L, 0.2mg/L, 0.4mg/L, 0.8mg/L, 1.6mg/L, the concentration The interval factor is 2. The solution update frequency is required to be no less than 12 hours (that is, all solutions in the test tank must be updated once within 12 hours). Environmental condition control must meet the requirements of Chemical Testing Guidelines (HJ/153-2004) and "Chemical Testing Methods" (Second Edition, China Environmental Publishing House). The implementation of the present invention is as follows:

(1) Turn on the constant temperature water bath, set the temperature at 20±1°C, turn on the light and set the photoperiod as 16h (light): 8h (dark).

(2) The procedure for setting the peristaltic pump is as follows:

Peristaltic pump 1

Program 1 (P1P1): channel 1 flow rate 0.50mL/min, forward run 40min

Program 2 (P1P2): channel 2 flow rate 1.00mL/min, reverse run 50min

Peristaltic pump 2

Program 1 (P2P1): channel 1 flow rate 100 mL/min, forward run 5.0 min

Program 2 (P2P2): channel 2 flow rate 100 mL/min, reverse run 5.0 min

Peristaltic pump 3

Program 1 (P3P1): channel 1 flow rate 100 mL/min, forward run 5.0 min

Program 2 (P3P2): channel 2 flow rate 100mL/min, reverse run 5.0 min

Peristaltic pump 4

Program 1 (P4P1): flow rate 100 mL/min, run 9.8 min

Program 2 (P4P2): flow rate 100 mL/min, run 9.5 min

Program 3 (P4P3): flow rate 100 mL/min, run for 5.0 min

Program 4 (P4P4): flow rate 100 mL/min, run for 5.0 min

Program 5 (P4P5): flow rate 100 mL/min, run for 5.0 min

Program 6 (P4P6): flow rate 100 mL/min, run for 5.0 min

Program 7 (P4P7): flow rate 100 mL/min, run for 10 min

Program 8 (P4P8): flow rate 100 mL/min, run for 10 min

Peristaltic pump 5

Program (P5P): flow rate 0.05 mL/min, forward run 2.0 min

Peristaltic pump 6

Program (P6P): flow rate 50.0 mL/min, run forward until the end of the test.

(3) Prepare 100mL 16g/L grade I mother liquor of the test substance, and put it into the grade I mother liquor bottle. At the same time, 120 L of Elendt M7 medium was prepared and loaded into a medium storage barrel. The amount of the test substance grade I mother solution is sufficient for the entire test period of 21 days. Unless the test substance is poor in stability, the test process does not need to be prepared again; the amount of the medium can supply the consumption of the 3-d test, so it only needs to be prepared once every 3 days .

(4) Start the PLC program in the PLC controller, and the system will begin to prepare the test solution (this process takes about 160 minutes). After the solution is prepared, it will automatically enter the solution temporary storage system (this process takes about 10 minutes), and then automatically and continuously pump it into In the test cylinders with different test concentrations (this process takes about 60 minutes), the test solution exceeding the capacity of the test cylinder is continuously discharged from the water outlet to the waste liquid treatment system, and the waste liquid treatment system performs filtration and adsorption treatment and then discharges the effluent. Therefore, a complete solution preparation and distribution process takes 250 minutes, and each concentration test cylinder solution can be renewed by 1.0L. Therefore, it takes up to 11.5 hours to complete the update of the 3L solution of the test tank (the actual time is less, because the second-level mother liquor does not need to be re-prepared every time, and the process takes 40 minutes for a single time), which meets the requirement of completing all test tank solution updates within 12 hours requirements.

(5) After the test tank is filled with the test solution, put the healthy and lively juvenile daphnia aged 6 to 8 hours into the small enclosures of the toxicity exposure system, and put one daphnia in each small enclosure. Cover the dome cover, and the test officially begins.

(6) During the test, the dome-shaped cover was opened every day to observe the survival and reproduction of the mother daphnia and the survival of the young daphnia, and the young daphnia were removed in time. At the same time, the green algae concentrate is added to the small shroud every day to feed the daphnia. During observation and feeding, the tray in the test tank can be rotated to move the observed small enclosure to a position that is easy for the observer to operate, and at the same time, record according to the numbers R1~R10 on the tray to avoid mistakes and omissions.

(7) When the solution in the solution temporary storage system is used up, the system automatically starts a new round of solution preparation and transmission process to ensure the continuous update of the solution until the end of the 21d test.

Claims (2)

1. a kind of Daphnia magna continuous-flow type toxicity test device, it is characterised in that：The experimental rig includes solution compounding system, molten Liquid scratch system, toxic exposure system, solution automatic cycle and make-up system, liquid waste treatment system；The solution compounding system Pass through solution automatic cycle and make-up system phase respectively between solution scratch system, solution scratch system and toxic exposure system Even, toxic exposure system is connected with liquid waste treatment system；

The solution compounding system includes 1 set of automatic liquid dispenser, 1 nutrient media storage bucket, 1 I grade of mother liquid multi-cavity bottle, 1 II grade of mother Liquid bottle, 1 cosolvent bottle, bottle is prepared in 1 cosolvent control, bottle is prepared in 1 blank control, 5 test solutions prepare bottle, II grade Mother liquid multi-cavity bottle, cosolvent control prepare bottle, bottle is prepared in blank control, test solution prepares bottle and is respectively placed in independent magnetic stirring apparatus On；

The automatic liquid dispenser includes 3 binary channel peristaltic pumps, 2 unipath peristaltic pumps, 1 liquid level sensor, 1 screw rod biography Motivation structure, 1 eight ways valve, a PLC controller；The bottom of the bottom of I grades of mother liquid multi-cavity bottles and II grade of mother liquor bottleneck, II grade of mother liquid multi-cavity bottle The bottleneck that bottle is prepared with first test solution is connected by two woven hoses of the same binary channel peristaltic pump；First tested The bottom of solution preparation bottle prepares the bottleneck of bottle with second test solution, second test solution prepares bottom and the third of bottle The bottleneck that a test solution prepares bottle is connected by two woven hoses of the same binary channel peristaltic pump；Third test solution is matched The bottom of bottle processed and the 4th test solution prepare the bottleneck of bottle, the 4th test solution prepare bottle bottom and the 5th it is tested The bottleneck of solution preparation bottle is connected by two woven hoses of the same binary channel peristaltic pump；Nutrient media storage bottom of the barrel passes through list Woven hose, screw drive mechanism and the eight ways valve of access peristaltic pump are compareed with II grade of mother liquid multi-cavity bottle, cosolvent respectively prepares bottle, blank Bottle is prepared in control, 5 test solutions prepare bottle；The liquid level sensor is placed in above II grade of mother liquid multi-cavity bottle；

The solution scratch system includes that bottle is kept in 1 cosolvent control, bottle is kept in 1 blank control, 5 test solutions are temporary Bottle, the entrances of above-mentioned 7 temporary bottles compare with cosolvent respectively prepares bottle, blank control preparation bottle and 5 solution preparation bottles Outlet is connected by pipeline, and pipeline is equipped with solenoid valve；All temporary bottles are brown glass material, and are equipped with head cover, head cover Apertured is inserted into for the woven hose of peristaltic pump；All reagent bottles are respectively placed on independent magnetic stirring apparatus, the temporary system of the solution System is placed in below solution compounding system；

The solution automatic cycle includes 2 liquid level sensors, 1 automatic electromagnetic valve, 1 seven accesses wriggling with make-up system Pump, 1 PLC controller, a liquid level sensor are mounted on the 5th test solution and prepare in bottle, another liquid level sensor peace It is kept in bottle mounted in the 5th solution；Bottle is prepared in cosolvent control, bottle is prepared in blank control and 5 solution preparation bottles pass through respectively Seven woven hoses of the seven accesses peristaltic pump compare temporary bottle with cosolvent, bottle is kept in blank control, 5 test solutions are temporary Accordingly independent connection, solenoid valve are mounted in the middle part of seven woven hoses bottle, block flow or unicom water automatically by PLC controller Stream；

The toxic exposure system includes that the illumination automatically controlled 1 constant temperature water bath, 1 group of photoperiod, 1 cosolvent control are surveyed It trying cylinder, 1 blank control test cylinder, 5 test solutions and tests cylinder, above-mentioned 7 tests cylinder is placed in constant temperature water bath, and described 7 The entrance of a test cylinder is connected with above-mentioned corresponding 7 temporary bottles by seven independent woven hoses of seven access peristaltic pumps；

The liquid waste treatment system includes that 1 layer of filter cotton and 1 layer of activated carbon Filter column, the two are integrated in closed system, only logical Water inlet pipe is crossed with each test cylinder apopore of toxic exposure system to be connected；

It is brown glass material that nutrient media storage bottom of the barrel in the solution compounding system, which has universal wheel, all reagent bottles, Matter；Nutrient media storage bucket and reagent bottle have head cover, head cover apertured to be inserted into for the woven hose of peristaltic pump；

The test cylinder is transparent glass material, matches head cover, and casing wall is equipped with inlet opening, apopore, and each test in cylinder is placed 10 small shrouds and a dismountable shroud frame, shroud are made of stainless (steel) wire, the mesh of 60 mesh of aperture ~ 80, upper end opening；Shroud Frame is the glass plate of trepanning, and hole count 10, aperture is the same as shroud outer diameter；Shroud frame antenna height slightly below tests cylinder mouth, and 10 are enclosed Cylinder is inserted into test cylinder from shroud frame opening, and shroud bottom is kept to be slightly above cylinder bottom, and shroud mouth is slightly above shroud frame；Inlet opening It is located at below shroud frame with apopore, wherein inlet opening is close to test cylinder bottom, and apopore is close to shroud frame, and inlet opening, water outlet Hole is located at the test casing wall both ends that cylinder is opposite, air line distance is farthest.

2. a kind of Daphnia magna continuous-flow type toxicity test device according to claim 1, it is characterised in that：The PLC controls The control method of device processed is as follows：

" beginning " key or the liquid level sensor above II grade of mother liquid multi-cavity bottle started in PLC controller sends out the signal less than warning value When, pump is added from draw solution in I grades of mother liquid multi-cavity bottles in II grade of mother liquid multi-cavity bottle；Call screw drive mechanism by eight ways valve outlet alignment II Grade mother liquid multi-cavity bottle from absorption M7 culture mediums in nutrient media storage bucket and is added in II grade of mother liquid multi-cavity bottle；After 20min from II grade of mother liquid multi-cavity bottle Draw solution is simultaneously added in first test solution preparation bottle, and eight ways valve outlet is directed at first test solution and prepares bottle, from training It supports to draw M7 culture mediums in base storage drum and first test solution is added and prepares bottle；Match from first test solution after 20min Second test solution of draw solution and addition is prepared in bottle in bottle processed, then is drawn M7 culture mediums from nutrient media storage bucket and added Enter second test solution and prepares bottle；After 20min, it is tested to prepare draw solution addition third in bottle from second test solution In solution preparation bottle, then from absorption M7 culture mediums in nutrient media storage bucket and it is added in third test solution preparation bottle；20min Draw solution in bottle is prepared from third test solution afterwards the 4th test solution is added and prepared in bottle, then from nutrient media storage bucket Middle absorption M7 culture mediums are simultaneously added in the 4th test solution preparation bottle；It prepares in bottle and inhales from the 4th test solution after 20min Take solution that the 5th test solution is added to prepare in bottle, then draw from nutrient media storage bucket M7 culture mediums and be added the 5th by It tries in solution preparation bottle；Then it prepares in bottle from drawing solvent in cosolvent bottle cosolvent control is added, then from nutrient media storage M7 culture mediums are drawn in bucket and cosolvent control is added prepares in bottle；M7 culture mediums are drawn from nutrient media storage bucket and sky is added White control is prepared in bottle；

When liquid level sensor above the temporary bottle of the 5th test solution sends out signal of the liquid level less than warning value, program will be electric Magnet valve is opened, and 5 solution preparation bottles, blank control prepare bottle, cosolvent control prepares the solution in bottle by gravity point It Jin Ru not the temporary bottle of 5 solution, the temporary bottle of blank control, the temporary bottle of cosolvent control；Liquid above the 5th solution preparation bottle When face sensor sends out signal of the liquid level less than warning value, program closes solenoid valve, prepares bottle and liquid barrier in temporary bottle； Then it is prepared in bottle from first test solution of draw solution in II grade of mother liquid multi-cavity bottle and addition, screw drive mechanism is called to lead to eight Valve outlet is directed at first test solution and prepares bottle, and M7 culture mediums are drawn from nutrient media storage bucket and addition first is tested molten Liquid prepares bottle；Draw solution in bottle is prepared from first test solution and be added in second test solution preparation bottle after 20min, M7 culture mediums are drawn from nutrient media storage bucket again and second test solution is added and prepare bottle；It is tested from second after 20min Draw solution is added third test solution and prepares in bottle in solution preparation bottle, then M7 culture mediums are drawn from nutrient media storage bucket And third test solution is added and prepares in bottle；Draw solution in bottle is prepared from third test solution be added the 4th after 20min A test solution is prepared in bottle, then draws M7 culture mediums from nutrient media storage bucket and the 4th test solution preparation bottle is added In；From the 4th test solution draw solution in bottle is prepared after 20min the 5th test solution is added and prepared in bottle, then from culture M7 culture mediums are drawn in base storage drum and the 5th test solution is added prepares in bottle；Then solvent is drawn from cosolvent bottle to add Enter cosolvent control to prepare in bottle, then is prepared in bottle from drawing M7 culture mediums in nutrient media storage bucket and cosolvent control being added； It is prepared in bottle from drawing M7 culture mediums in nutrient media storage bucket and blank control being added；

Solution keeps in bottle from solution incessantly by seven access peristaltic pumps and is pumped into test cylinder, and the test beyond test cylinder capacity is molten Liquid is constantly discharged to liquid waste treatment system from water outlet.