CN202664020U - Daphnia magna culturing and separating device - Google Patents
Daphnia magna culturing and separating device Download PDFInfo
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- CN202664020U CN202664020U CN 201220297173 CN201220297173U CN202664020U CN 202664020 U CN202664020 U CN 202664020U CN 201220297173 CN201220297173 CN 201220297173 CN 201220297173 U CN201220297173 U CN 201220297173U CN 202664020 U CN202664020 U CN 202664020U
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Abstract
The utility model discloses a daphnia magna culturing and separating device. The daphnia magna culturing and separating device comprises an alga growth promoting device, a daphnia magna culturing container which is arranged in the alga growth promoting device, a circulating water device which is arranged in the daphnia magna culturing container, and a daphnia magna separating device. The daphnia magna culturing and separating device is easy and convenient to use, high in operability, simple in structure and convenient to manufacture and maintain, and the used materials are easily obtained. The culturing container is molded at one step and is small in size, so that the culturing container is prevented from being polluted by other materials, is easy to replace and facilitates subculturing; a flea-food growth promoting device facilitates the growth and reproduction of flea-food algae, so that the daphnia magna are not required to be fed every day; by the adoption of the circulating water device, aeration which is required in the process of culturing fleas is not required; and the separating device can prevent the fleas from leaving a water surface in the separation process, so that a large number of fleas can be separated quickly. By the adoption of the daphnia magna culturing and separating device, the quality of flea toxicity tests is guaranteed.
Description
Technical field
The utility model relates to the method for culturing and separating of zooplankton, is specifically related to a kind of culture of isolated device of Daphnia magna.
Background technology
The flea class belongs to the Arthropoda of invertebrate, Crustachia, and peg graft is the important monoid of freshwater zooplankton, is the bait of many Important Economic fish, simultaneously the self-cleaning in waters is had the very important effect.Daphnia magna has that life cycle is short, breeding is fast, economical, conveniently be easy to get, responsive and be easy in advantages such as laboratory cultures to poisonous substance, add their importance in aquatic ecosystem, thereby obtain the application of numerous countries, become a kind of code test indicator organism, be widely used for the aquatile toxicological test.
The U.S., European Union, Japan and other countries and international organization have set up ripe agricultural chemicals to flea class testing method at present, and China has also proposed flea class test method in 2004 in " chemical pesticide environmental safety assessment test rule " (declaration original text).
In these method of testings, test organism flea class has only been done the good condition of culture of maintenance, make Daphnia magna be in the parthenogenesis state, select and cultivate under the experiment condition more than 3 generations, the young flea of birth 6 ~ 24h, the children flea must bouncing, and the flea class can not be left this type of description of the water surface in the process of the test.Yet how to make the flea class be in the parthenogenesis state, how guarantee test is in 6 ~ 24h age with flea, and how the flea class is not left the water surface and do not specified in the guarantee test process.The domestic unit that respectively is engaged in flea class test do not have specifically yet can reference method come the guarantee test quality.Especially experimental amount is large, when large to flea class demand more without method simple and easy to operate.Scholar B. rood once was studied the cultural method of test with Daphnia magna, and the method exists can't satisfy the problem of a large amount of culture of isolated fleas.The patent No. of the announcements such as Huang Jianrong is that " a kind of Daphnia magna deposit conservation cultural method " patent of 201110047304 has mainly been set up simple flea-algae-spiral shell-fish and water and given birth to the ecosystem and lay in conservation, aquatile easily catches an illness in the actual mechanical process, must rebuild this system after catching an illness, quality of experiments can not be guaranteed.
The utility model content
Technical problem to be solved in the utility model provides a kind of culture of isolated device that guarantees the Daphnia magna of flea class test mass, this device can make Daphnia magna guarantee to reach under the manual control condition and cultivate more than 3 generations, the young flea of birth 6 ~ 24h is in the parthenogenesis state, and guarantee test during Young flea does not leave the water surface, better estimates the toxicity that agricultural chemicals produces aquatile flea class.
For solving the problems of the technologies described above, the technical solution adopted in the utility model is as follows:
A kind of culture of isolated device of Daphnia magna, it comprises that algal grown promotes device, places algal grown to promote the Daphnia magna culture vessel of device, places the circulating water device in the Daphnia magna culture vessel, and the flea separator;
Wherein, described algal grown promotes that device comprises, by the flaggy that the support upper end is supported, is fixed on the LED plant growth lamp of flaggy lower surface.
Wherein, described Daphnia magna culture vessel is that bottom and sidewall seal, the container of open-top.
Wherein, described flea separator is the logical cylinder of sidewall sealing, end face and bottom surface opened, the fixing two-layer mesh screen of logical cylinder internal activity, and the mesh diameter of upper strata mesh screen is 1mm, can guarantee that female flea can't pass but 6 ~ 24h children flea and flea kind can be passed through; The mesh diameter of lower floor's mesh screen is 0.5mm, can guarantee 6 ~ 24h children flea and the flea kind can't pass but water can pass through, and can reach the purpose of separating young flea by this flea separator.
Wherein, described circulating water device comprises the water inlet that is arranged on duct bottom, is arranged on the delivery port at pipeline top, and the suction pump that is connected with described pipeline, and described circulating water device is fixed on Daphnia magna culture vessel inwall.The diameter of pipeline need be enough to guarantee that maximum individual Daphnia magna passes through smoothly, and guarantees that delivery port is higher than the liquid level in the Daphnia magna culture vessel, and water inlet is lower than the liquid level in the Daphnia magna culture vessel.
Wherein, described LED plant growth lamp is right against the open top of Daphnia magna culture vessel, and area covers the opening at Daphnia magna culture vessel top substantially.
Wherein, the transversal size of described flea separator is less than the transversal size of Daphnia magna culture vessel, so that when needs separate Daphnia magna, can place in the Daphnia magna culture vessel.
Wherein, described flea separator transversal is of a size of half of transversal size of Daphnia magna culture vessel.
Wherein, be provided with 2 ~ 4 suckers bottom the described flea separator sidewall.
Wherein, the logical cylinder of described flea separator both sides, top can be arranged with two rods, make things convenient for taking of separator.
Wherein, in the described flea separator, upper strata mesh screen and lower floor's mesh screen can take out from logical cylinder, also can be fixed in the logical cylinder, for example, can adopt shackle designs, mesh screen is hung on logical cylinder inwall, perhaps adopt flexible member, the mesh screen frame is squeezed in the interior rib groove of logical cylinder, all can, those skilled in the art can rely on the activity of existing techniques in realizing mesh screen be fixed in the function of logical cylinder inwall.
Wherein, in the described flea separator, the net in the mesh screen is soft net, can adopt prior art net to be fixed in the frame of mesh screen, for example technology of suturing skill, spiral technique for fixing or two frame compression etc.
Wherein, the pipeline of the described circulating water device outside is provided with geometrical clamp, and geometrical clamp can be fixed on the sidewall of Daphnia magna culture vessel.
Wherein, described support material is stainless steel; Flaggy (6) material is timber; The material of Daphnia magna culture vessel is glass, and the glass one-shot forming is without binding material; The pipeline of circulating water device is environmentally-friendly plastic; The side of flea separator and mesh screen side material are stainless steel.
Utilize said apparatus to carry out the method for culturing and separating of Daphnia magna, the method comprises the steps:
A under 20 ± 2 ℃ of indoor conditionss, injected water in the Daphnia magna culture vessel, the water surface is higher than the water inlet of circulating water device, be lower than the delivery port of circulating water device, operating circulating water device 1 day continues to add the algae food of Daphnia magna in the Daphnia magna culture vessel, food concentration reaches 10
5Individual/mL, operating circulating water device 10 clocks;
B continues to add the Daphnia magna kind in the Daphnia magna culture vessel, open algal grown and promote LED plant growth lamp on the device, in the process of growth of flea every day timing operation circulating water device 1 hour, system's operation 3 days;
C places the Daphnia magna culture vessel with the flea separator, so that the water surface is lower than the sidewall top of flea separator, and ON cycle water device, so that the delivery port of circulating water device is positioned at the top of flea separator, water outlet can directly fall in the flea separator; Remove the upper strata screen cloth, keep lower screen, use the basin greater than the cross section of flea separator, underwater in the Daphnia magna culture vessel places the flea separation unit bottoms with described basin, when guaranteeing that the flea separator takes out the Daphnia magna culture vessel, lower screen still is immersed in below the interior water surface of basin;
D takes out basin with the flea separator, place in advance in the other cover Daphnia magna culture vessel of processing according to the mode of steps A, concussion, so that the Daphnia magna on the lower screen can fall into the water, shift out basin and flea separator, open algal grown and promote LED plant growth lamp on the device, in the process of growth of flea every day timing operation circulating water device 1 hour, system's operation 3 days;
F is cycled to repeat step C and step D, moves the 9th day to system and finish to turn out first generation children flea from the flea kind; Turned out afterwards generation children flea every 9 days, until turn out the 4th generation children flea, utilize the flea separator to separate, the young flea that takes off on layer screen cloth is used for flea class toxicity test.
By method of the present utility model, can guarantee finally to obtain the 4th generation, birth 6 ~ 24h and be in the young flea of parthenogenesis state, guaranteed simultaneously in the process of the test not leaving water of flea class.
Among the step B, described algae food is chlorella or grid algae.
Among the step B, described LED plant growth lamp (5) is only opened at night by timing means control, moves than 16:8 according to the dark time of light.
The method for culturing and separating of Daphnia magna of the present utility model needs the culture of isolated device of at least two cover Daphnia magnas, uses by turns, perhaps uses at least two cover Daphnia magna culture vessels (2) to be placed on by turns algal grown and promotes device, is used in conjunction with circulating water device.
Flea separator of the present utility model only just uses when needs separate Daphnia magna, is placed in the Daphnia magna culture vessel; When cultivating Daphnia magna, do not need to be placed in the Daphnia magna culture vessel.
Beneficial effect:
1, the utility model is simple and easy to do, and is workable.The utility model material therefor is easy to obtain, simple in structure, manufacturing and easy to maintenance.
2, the utility model adopts Daphnia magna culture vessel once-forming and volume little, has avoided the pollution of other material and be easy to changing, and makes things convenient for subculture to cultivate; Flea group food growth promoting motion device is beneficial to the growth and breeding of flea group food algae, saves the work of feeding every day; It is rapid that circulating water device has reduced sudden and violent jia bombardier required in the flea class incubation; Separator has been avoided leaving the water surface in the separation process of flea class, has guaranteed that a large amount of of flea class separate fast.
3, after the off-test, keep two groups of culture apparatuses, rotating at regular intervals feeds gets final product.Remove the full-time conservation work of special messenger from.
Description of drawings:
Fig. 1 is that algal grown of the present utility model promotes the apparatus structure schematic diagram.
Fig. 2 is Daphnia magna culture vessel structural representation of the present utility model.
Fig. 3 is flea separator structural representation of the present utility model.
Fig. 4 is circulating water device structural representation of the present utility model.
Fig. 5 is each component combined structure schematic diagram of culture of isolated device of Daphnia magna of the present utility model.
Wherein, 1 algal grown promotes device; 2 Daphnia magna culture vessels; 3 flea separators; 4 circulating water devices; The 5LED plant growth lamp; 6 flaggies; 7 supports; 8 upper strata mesh screens; 9 lower floor's mesh screens; 10 suckers; 11 water inlets; 12 delivery ports; 13 geometrical clamps; 14 suction pumps.
Embodiment
According to following embodiment, can understand better the utility model.Those skilled in the art will readily understand, embodiment only is used for explanation the utility model, and should also can not limit the utility model described in detail in claims.
Embodiment 1:
As shown in Figure 5, the culture of isolated device of Daphnia magna of the present utility model, it comprises that algal grown promotes device 1, places algal grown to promote the Daphnia magna culture vessel 2 of device 1, place the circulating water device 4 in the Daphnia magna culture vessel 2, and flea separator 3.
Wherein, as shown in Figure 1, algal grown promotes that device 1 comprises, by the flaggy 6 that support 7 upper ends are supported, is fixed on the LED plant growth lamp 5 of flaggy 6 lower surfaces.Flaggy 6 materials are preferably timber.Described support 7 materials are preferably stainless steel.For better implementing the utility model, described LED plant growth lamp 5 is right against the top that Daphnia magna culture vessel 2 is opened, and area covers the opening at Daphnia magna culture vessel 2 tops substantially.LED plant growth lamp 5 is only opened at night by timing means control, according to the dark time of light than 16:8 operation (be to open 16h daytime, closed night 8 hours).
Wherein, as shown in Figure 2, Daphnia magna culture vessel 2 is the container of the sealing of bottom and sidewall, open-top, and the material of Daphnia magna culture vessel 2 is preferably glass, and the glass one-shot forming is without binding material, the preferred 0.5cm of wall thickness.
Wherein, as shown in Figure 3, flea separator 3 is the logical cylinder of sidewall sealing, end face and bottom surface opened, the fixing two-layer mesh screen of logical cylinder internal activity, and the mesh diameter of upper strata mesh screen is 1mm, can guarantee that female flea can't pass but 6 ~ 24h children flea can pass through; The mesh diameter of lower floor's mesh screen is 0.5mm, can guarantee that 6 ~ 24h children flea can't pass but water can pass through, and can reach the purpose of separating young flea by this flea separator.The side of flea separator 3 and mesh screen side material are stainless steel.For better realizing the utility model, the transversal size of described flea separator 3 is less than the transversal size of Daphnia magna culture vessel 2, so that when needs separate Daphnia magna, can place in the Daphnia magna culture vessel 2.Preferably, described flea separator 3 transversal is of a size of half of transversal size of Daphnia magna culture vessel 2, and for example Daphnia magna culture vessel 2 is the cylinder of 40cm for cross-sectional diameter, and flea separator 3 is preferably the cylinder of cross section 20cm.Described flea separator 3 sidewalls bottom is provided with 2 ~ 4 suckers, the flea separator can be fixed in the Daphnia magna culture vessel 2, can not rock because of the variation of current.
Wherein, as shown in Figure 4, circulating water device 4 comprises the water inlet 11 that is arranged on duct bottom, is arranged on the delivery port 12 at pipeline top, and the suction pump 14 that is connected with described pipeline.Described pipeline is environmentally-friendly plastic, the diameter of pipeline need be enough to guarantee that maximum individual Daphnia magna passes through smoothly, preferred caliber 1cm, and guarantee that delivery port is higher than the liquid level in the Daphnia magna culture vessel, water inlet are lower than liquid level in the Daphnia magna culture vessel near the Daphnia magna bottom of culture vessel.Described circulating water device is fixed on Daphnia magna culture vessel 2 inwalls.For better implementing the utility model, the pipeline outside of described circulating water device 4 is provided with geometrical clamp 13, and geometrical clamp can be fixed on the sidewall of Daphnia magna culture vessel 2.
Embodiment 2:
The device of a kind of embodiment of utilization 1 carries out the method for culturing and separating of Daphnia magna, and the method comprises the steps:
A is under 20 ± 2 ℃ of indoor conditionss, and to Daphnia magna culture vessel 2 interior injection 15L water, the water surface is higher than the water inlet of circulating water device 4, be lower than the delivery port of circulating water device 4, operating circulating water device 41 days, this process are equivalent to sudden and violent gas process, but can not change water pH value.Continuation adds the algae food of Daphnia magna in the Daphnia magna culture vessel 2, and such as chlorella or grid algae etc., food concentration reaches 10
5Individual/mL, operating circulating water device 410 clocks.
B continues to Daphnia magna culture vessel 2 interior adding Daphnia magna kinds, open algal grown and promote LED plant growth lamp 5 on the device 1, in the process of growth of flea every day timing operation circulating water device 1 hour, system's operation 3 days.
C places Daphnia magna culture vessel 2 with flea separator 3, so that the water surface is lower than the sidewall top of flea separator 3,4 half an hour of ON cycle water device, so that the delivery port of circulating water device 4 is positioned at the top of flea separator 3, water outlet can directly fall in the flea separator 3; Remove upper strata screen cloth 8, keep lower screen 9, use the basin greater than the cross section of flea separator 3, underwater in Daphnia magna culture vessel 2 places flea separator 3 bottoms with described basin, when guaranteeing that flea separator 3 takes out Daphnia magna culture vessel 2, lower screen 9 still is immersed in below the interior water surface of basin.
D takes out basin with flea separator 3, place in advance in the other cover Daphnia magna culture vessel 2 of processing according to the mode of steps A, slight concussion, so that the Daphnia magna on the lower screen 9 can fall into the water, shift out basin and flea separator 3, open algal grown and promote LED plant growth lamp 5 on the device 1, in the process of growth of flea every day timing operation circulating water device 1 hour, system's operation 3 days.
F is cycled to repeat step C and step D, moves the 9th day to system and finish to turn out first generation children flea from the flea kind; Turned out afterwards generation children flea every 9 days, until turn out the 4th generation children flea, utilize the flea separator to separate, the young flea that takes off on layer screen cloth is used for flea class toxicity test.
By method of the present utility model, can guarantee finally to obtain the 4th generation, birth 6 ~ 24h and be in the young flea of parthenogenesis state, guaranteed simultaneously in the process of the test not leaving water of flea class.
Embodiment 3: the test of flea class reference material.
The Daphnia magna acute toxicity experience is used for estimating tested material to the issuable impact of aquatile.Under rated condition, make the aqueous solution of Daphnia magna contact variable concentrations tested material, the tested material concentration when determining Daphnia magna dead 50%.This test determination the acute toxicity of reference material potassium bichromate to Daphnia magna, whether biological and experimental condition meets the requirements with confirmed test.
1 materials and methods
1.1 for examination Daphnia magna kind
Daphnia magna (Daphnia magna Straus), the flea kind is raised and train by this laboratory.Experiment obtains with the method for flea according to embodiment 2.
1.2 experimental condition
Instrument and equipment: balance, dissolved oxygen meter, thermometer, pH meter, pipettor, glassware etc.
Test water parameter: pH 7.49 ~ 7.55, dissolved oxygen 8.85 ~ 8.98mg/L.19.7 ~ 19.9 ℃ of test water temperatures.
1.3 test method
In the 100mL beaker, pack into the during test potassium bichromate solution of 50mL variable concentrations, every beaker is put into 5 for the young fleas of examination, and young flea flea age is 6 ~ 24h, in the movable downtrod young flea number of 24h observed and recorded.Not bait throwing in of duration of test material.
2 results
Result's demonstration, potassium bichromate is to the EC of Daphnia magna 24h
50For 1.08mg/L(sees Table 1).
3 conclusions
Potassium bichromate is to the EC of Daphnia magna 24h
50Be 1.08mg/L, meet " chemical pesticide environmental safety assessment test rule " middle potassium bichromate to Daphnia magna 24h-EC
50It is the requirement of 0.6 ~ 1.7mg/L.
Table 1 potassium bichromate is to the toxicity of Daphnia magna
Claims (7)
1. the culture of isolated device of a Daphnia magna, it is characterized in that, it comprises that algal grown promotes device (1), places algal grown to promote the Daphnia magna culture vessel (2) of device (1), place the circulating water device (4) in the Daphnia magna culture vessel (2), and flea separator (3);
Wherein, described algal grown promotes device (1) to comprise, by the flaggy (6) that support (7) upper end is supported, is fixed on the LED plant growth lamp (5) of flaggy (6) lower surface;
Described Daphnia magna culture vessel (2) is that bottom and sidewall seal, the container of open-top;
Described flea separator (3) is the logical cylinder of sidewall sealing, end face and bottom surface opened, the fixing two-layer mesh screen of logical cylinder internal activity, and the mesh diameter of upper strata mesh screen (8) is 1mm, the mesh diameter of lower floor's mesh screen (9) is 0.5mm;
Described circulating water device (4) comprises the water inlet (11) that is arranged on duct bottom, be arranged on the delivery port (12) at pipeline top, and the suction pump (14) that is connected with described pipeline, described circulating water device is fixed on Daphnia magna culture vessel (2) inwall.
2. the culture of isolated device of Daphnia magna according to claim 1 is characterized in that, described LED plant growth lamp (5) is right against the open top of Daphnia magna culture vessel (2), and area covers the opening at Daphnia magna culture vessel (2) top substantially.
3. the culture of isolated device of Daphnia magna according to claim 1, it is characterized in that, the transversal size of described flea separator (3) is less than the transversal size of Daphnia magna culture vessel (2), so that when needs separate Daphnia magna, can place in the Daphnia magna culture vessel (2).
4. the culture of isolated device of Daphnia magna according to claim 1 is characterized in that, described flea separator (3) transversal is of a size of half of transversal size of Daphnia magna culture vessel (2).
5. the culture of isolated device of Daphnia magna according to claim 1 is characterized in that, described flea separator (3) sidewall bottom is provided with 2 ~ 4 suckers.
6. the culture of isolated device of Daphnia magna according to claim 1 is characterized in that, the pipeline outside of described circulating water device (4) is provided with geometrical clamp (13), and geometrical clamp can be fixed on the sidewall of Daphnia magna culture vessel (2).
7. the culture of isolated device of Daphnia magna according to claim 1 is characterized in that, described support (7) material is stainless steel; Flaggy (6) material is timber; The material of Daphnia magna culture vessel (2) is glass, and the glass one-shot forming is without binding material; The pipeline of circulating water device (4) is environmentally-friendly plastic; The side of flea separator (3) and mesh screen side material are stainless steel.
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| CN 201220297173 CN202664020U (en) | 2012-06-21 | 2012-06-21 | Daphnia magna culturing and separating device |
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| CN 201220297173 CN202664020U (en) | 2012-06-21 | 2012-06-21 | Daphnia magna culturing and separating device |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102687709A (en) * | 2012-06-21 | 2012-09-26 | 环境保护部南京环境科学研究所 | Culture and separation device and culture and separation method for daphnia magna |
| CN103598150A (en) * | 2013-11-30 | 2014-02-26 | 哈尔滨工业大学 | Rapid cultivation device and method of daphnia magna laboratory |
| CN114431179A (en) * | 2022-02-25 | 2022-05-06 | 生态环境部南京环境科学研究所 | Evaluation method for safety influence of Bt insecticidal protein on daphnia magna |
-
2012
- 2012-06-21 CN CN 201220297173 patent/CN202664020U/en not_active Expired - Fee Related
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102687709A (en) * | 2012-06-21 | 2012-09-26 | 环境保护部南京环境科学研究所 | Culture and separation device and culture and separation method for daphnia magna |
| CN102687709B (en) * | 2012-06-21 | 2013-12-11 | 环境保护部南京环境科学研究所 | Culture and separation device and culture and separation method for daphnia magna |
| CN103598150A (en) * | 2013-11-30 | 2014-02-26 | 哈尔滨工业大学 | Rapid cultivation device and method of daphnia magna laboratory |
| CN103598150B (en) * | 2013-11-30 | 2015-12-02 | 哈尔滨工业大学 | Daphnia magna laboratory fast culture device and cultural method |
| CN114431179A (en) * | 2022-02-25 | 2022-05-06 | 生态环境部南京环境科学研究所 | Evaluation method for safety influence of Bt insecticidal protein on daphnia magna |
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