CN103598150A - Rapid cultivation device and method of daphnia magna laboratory - Google Patents
Rapid cultivation device and method of daphnia magna laboratory Download PDFInfo
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- CN103598150A CN103598150A CN201310624588.3A CN201310624588A CN103598150A CN 103598150 A CN103598150 A CN 103598150A CN 201310624588 A CN201310624588 A CN 201310624588A CN 103598150 A CN103598150 A CN 103598150A
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- 241001494246 Daphnia magna Species 0.000 title claims abstract description 87
- 238000000034 method Methods 0.000 title claims abstract description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 76
- 238000010438 heat treatment Methods 0.000 claims abstract description 26
- 238000012360 testing method Methods 0.000 claims abstract description 19
- 238000005273 aeration Methods 0.000 claims description 22
- 239000011521 glass Substances 0.000 claims description 13
- 240000009108 Chlorella vulgaris Species 0.000 claims description 9
- 235000007089 Chlorella vulgaris Nutrition 0.000 claims description 9
- 235000013305 food Nutrition 0.000 claims description 8
- 239000004744 fabric Substances 0.000 claims description 7
- 238000011081 inoculation Methods 0.000 claims description 7
- 238000005286 illumination Methods 0.000 claims description 6
- 241000195493 Cryptophyta Species 0.000 claims description 4
- 238000009395 breeding Methods 0.000 claims description 4
- 230000001488 breeding effect Effects 0.000 claims description 4
- 125000001309 chloro group Chemical group Cl* 0.000 claims description 4
- 239000012531 culture fluid Substances 0.000 claims description 3
- 241000251468 Actinopterygii Species 0.000 claims description 2
- 229910000746 Structural steel Inorganic materials 0.000 claims description 2
- 230000003203 everyday effect Effects 0.000 claims description 2
- 239000000463 material Substances 0.000 abstract description 7
- 231100000331 toxic Toxicity 0.000 abstract 1
- 230000002588 toxic effect Effects 0.000 abstract 1
- 230000035558 fertility Effects 0.000 description 4
- 230000008186 parthenogenesis Effects 0.000 description 4
- 231100000820 toxicity test Toxicity 0.000 description 3
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000010561 standard procedure Methods 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 241000238421 Arthropoda Species 0.000 description 1
- 241000252254 Catostomidae Species 0.000 description 1
- 241000238571 Cladocera Species 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000033001 locomotion Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000012423 maintenance Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000029264 phototaxis Effects 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 230000007096 poisonous effect Effects 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
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Abstract
The invention provides a rapid cultivation device and method of a daphnia magna laboratory and belongs to the field of cultivation of zooplankter. The rapid cultivation device comprises supports, base plates, water bath tanks, a water storage tank, cultivation cups, heating rods, thermometers, turbulence pumps, a fluorescent lamp and a time controller. Each base plate is arranged at the bottom of the corresponding support. The storage water tank is arranged on the base plate of the bottommost support. The water bath tanks are arranged on the base plates of the two upper supports. One thermometer, one turbulence pump and two heating rods are arranged in the water storage tank. Each water bath tank is provided with one thermometer, two heating rods and six cultivation cups. The fluorescent lamp is located above the portion deviating from the center of the support and is parallel to the support. The time controller is in contact with the fluorescent lamp. The rapid cultivation device is easy and convenient to operate, high in operability, easy to set up and free of a complicated device structure. Daphnia magna cultivated through the rapid cultivation device is rapid in reproduction and large in quantity, the life condition of the daphnia magna is stable, and the individual difference of the daphnia magna is small, so that results of toxic tests of fleas are stable and reliable, the test quality is guaranteed, and enough test materials can be provided for tests such as removal of zooplankter pollution to water in the industry of water treatment at the same time.
Description
Technical field
The invention belongs to zooplankton and cultivate field, relate to a kind of culture apparatus and cultural method thereof of Daphnia magna.
Background technology
Daphnia magna (Daphnia magna) belongs to Arthropoda, Crustachia, cladocera, is a kind of common freshwater zooplankton, has critical role in aquatic ecosystem.Because it has widely distributedly, life cycle is short, and reproduction speed is fast, and reproduction amount is high, is easy to cultivate, and the features such as poisonous substance sensitivity are widely used in to Aquatic Toxicity Test.
Daphnia magna is internationally recognized code test indicator organism at present, and many countries have also set up the standard method of Daphnia magna toxotest simultaneously, and the condition of culture of test Daphnia magna has been done to relevant regulation.But the Daphnia magna in the most of laboratories of China is cultivated and lacked effective condition of culture or device at present, is difficult to guarantee culture effect, and this makes the culture environment of Daphnia magna stable not, Daphnia magna is difficult to the long-term parthenogenesis that keeps, and Daphnia magna quantity is difficult to guarantee.Therefore be difficult to obtain the required Daphnia magna of test, such as Daphnia magna is different the length of time, individuality is neat not, and life state is not equal, and these all can produce larger impact to result of the test, and test mass can not be guaranteed.
Summary of the invention
The object of the invention is to overcome the defect of existing Daphnia magna culture technique, a kind of method that improved laboratory Daphnia magna culture apparatus is provided and utilizes this Daphnia magna culture apparatus to carry out Daphnia magna cultivation.This device can be under manual control condition, realize the rapid Steady breed of Daphnia magna, and for a long time in parthenogenesis state, individual evenly neat, life state is active, can guarantee the quality of Daphnia magna toxicity test, and more simple than standard method, meet the requirement of laboratory fast culture and use.
Daphnia magna of the present invention laboratory fast culture device comprises support, base plate, cleansing bath tub, Water-accumulating water cylinder, cultivation cup, heating rod, thermometer, aeration pump, fluorescent lamp, time controller; The bottom of described support is provided with base plate, is placed with Water-accumulating water cylinder and cleansing bath tub on base plate, is provided with thermometer, aeration pump and heating rod in Water-accumulating water cylinder, and cleansing bath tub is provided with thermometer, heating rod and cultivation cup; Described fluorescent lamp is positioned at support oblique upper parallel with support, and time controller is connected with fluorescent lamp.
In said apparatus, described support is that angle iron builds, and is divided into three layers, and every layer height is adjustable, and the bottom is placed a Water-accumulating water cylinder, places two cleansing bath tubs for upper two-layer every layer.
In said apparatus, the bottom of every layer of described support is provided with base plate, this base plate can be wooden can be also glass system, guarantee to there is enough bearing capacities.
In said apparatus, described cleansing bath tub is glass system, square, at cleansing bath tub sidewall, near bottom, with sucker, heating rod is fixed in cleansing bath tub, and each cleansing bath tub is provided with a thermometer, two heating rods, six cultivation cups.
In said apparatus, described Water-accumulating water cylinder is glass system, square, at Water-accumulating water cylinder sidewall, near bottom, with sucker, heating rod is fixed in Water-accumulating water cylinder, and each Water-accumulating water cylinder is provided with a thermometer, an aeration pump, two heating rods.
In said apparatus, described cultivation cup be can splendid attire 2000ml water round glassware, one-shot forming non-cohesive material.
In said apparatus, described heating rod is the family expenses that can roughly the regulate temperature heating rod of breeding fish.
In said apparatus, described thermometer indication cleansing bath tub and Water-accumulating water cylinder temperature, coordinate heating rod to regulate Daphnia magna cultivation temperature.
In said apparatus, described aeration pump continues uninterrupted aeration to water in Water-accumulating water cylinder, to remove chlorine residue in water.
In said apparatus, 2 ~ 3 meters of described fluorescent lamp distance bracket oblique uppers, and parallel with support.
In said apparatus, described time controller links with fluorescent lamp, with the controlled light time.
Utilize said apparatus to cultivate a cultural method for Daphnia magna, concrete steps are as follows:
(1) running water is poured into cleansing bath tub and Water-accumulating water cylinder, aeration pump is opened, Water-accumulating water cylinder running water is carried out to continuous aeration, to remove chlorine residue in running water, utilize heating rod and thermometer that water temperature in cleansing bath tub and Water-accumulating water cylinder is adjusted to 20 ± 1 ℃ simultaneously;
(2) water in the above Water-accumulating water cylinder of aeration three days is added to and is cultivated in cup, every glass of 2000ml inoculates purebred, firm the be born Daphnia magna children flea of 6 ~ 24h in cultivating cup, and each cultivates 70 to 80 of cup inoculations, simultaneously the record inoculation date;
(3) chlorella vulgaris that does not contain medium is diluted in and is cultivated in cup, make to cultivate cup and present light green;
(4) adjust the position of fluorescent lamp, making to cultivate cup place intensity of illumination is 800 ~ 1000lux;
(5) between culture period, within every 3 days, change and cultivate the medium-and-large-sized flea culture fluid of cup, each 500ml that changes, the water that changes is in Water-accumulating water cylinder, aeration is more than three days, and temperature is the water of 20 ± 1 ℃, removes simultaneously and is deposited on Daphnia magna shelling, algae group and the dead Daphnia magna of cultivating cup bottom, append afterwards food, make to cultivate cup and present light green;
(6) when Daphnia magna produces young flea, with the separated female flea of screen cloth and the young flea in suitable aperture, before superseded 16 ages in days, female flea produces young flea, and female flea is put back to former cultivation cup and continues to cultivate;
(7) collect 16 ~ 22 ages in days mother 6 ~ 24h that flea produces children fleas and test or be inoculated into continuation cultivation in another one cultivation cup, and record date;
(8) treat that Daphnia magna surpasses 22 ages in days and can eliminate.
Can obtain according to the method described above cultivating Daphnia magnas more than 3 generations, and be all the female flea of 16-22 age in days and produce young flea, their quantity is large, evenly individual, fast growth, and fertility is strong, and life state is stablized.
The present invention has following beneficial effect:
(1) simple, the manufacture and easy to maintenance of Daphnia magna provided by the invention laboratory fast culture apparatus structure, expense is few, and material source is wide, easily builds.And can rationally utilize laboratory solid space, reduce floor space.
(2) Daphnia magna provided by the invention laboratory fast culture process can be under indoor conditions be cultivated all the year round, and Daphnia magna breeding amount is large, fast growth, and for Daphnia magna toxotest provides sufficient at any time, test material that can guarantee test quality.In addition, because Daphnia magna quantity is large, can be in water treatment field, remove in water the tests such as zooplankton enough test materials are provided.
(3) Daphnia magna provided by the invention laboratory fast culture process is without frequently changing water, and each quantity of exchanged water is few, has greatly reduced workload.
(4) Daphnia magna provided by the invention laboratory fast culture process selection chlorella vulgaris is Daphnia magna food, because chlorella vulgaris suspension is good, is difficult for precipitation, can make food be dispersed in Daphnia magna and cultivate in cup, is beneficial to Daphnia magna and filters food.Can avoid green alga to be attached on Daphnia magna feeler, thereby affect Daphnia magna locomitivity, the unfavorable problems such as life state and fertility simultaneously.
(5) this cultural method can make Daphnia magna photoenvironment stable, prevents that Daphnia magna local density is excessive, makes to cultivate in cup female flea density stabilized, keeps parthenogenesis state constantly.
Accompanying drawing explanation
Fig. 1 is Daphnia magna laboratory cultures device three-dimensional icon intention;
Fig. 2 is Daphnia magna laboratory cultures device front view schematic diagram;
Fig. 3 is Daphnia magna laboratory cultures device end view schematic diagram.
Embodiment
Below in conjunction with accompanying drawing, the present invention is described in further details.The present embodiment is only explanation of the invention; it is not limitation of the present invention; those skilled in the art can make to the present embodiment the modification that there is no creative contribution as required after reading this specification, but as long as within the scope of claim of the present invention, are all subject to the protection of Patent Law.
Daphnia magna laboratory cultures device as Figure 1-3, it comprises support 1, base plate 2, cleansing bath tub 3, Water-accumulating water cylinder 4, cultivates cup 5, heating rod 6, thermometer 7, aeration pump 8, fluorescent lamp 9, time controller 10.
This installs the title of each main member, and material and specification are as follows:
Support 1: iron, long 120cm, wide 40cm, high 150cm.Be divided into the 30cm of the San Ceng， the superiors, intermediate layer and orlop 55cm.
Base plate 2: wooden, also can glass system, there is enough bearing capacities, long 120cm, wide 40cm.
Cleansing bath tub 3: glass system, long 55cm, wide 30cm, high 20cm.
Water-accumulating water cylinder 4: glass system, long 110cm, wide 30cm, high 20cm.
Cultivate cup 5: glass system, disposal molding, without binder, circle, diameter 14cm, high 20cm, also available 2000ml beaker replaces.
Heating rod 6:100w, temperature is adjustable 18-32 ℃.
Thermometer 7: can survey 20 ± 1 ℃.
Aeration pump 8:5w.
Fluorescent lamp 9: power 40w.
Time controller 10: able to programme.
Support 1 is adjusted according to described every layer height, base plate 2 is placed on every layer of support 1.Upper two-layer placement cleansing bath tub 3, two every layer, orlop is placed a Water-accumulating water cylinder 4.Heating rod 6 is placed in to cleansing bath tub 3 and Water-accumulating water cylinder 4 sidewalls near bottom position, and two suckers of each heating rod 6 use are fixed, and each cleansing bath tub 3 and Water-accumulating water cylinder 4 are placed two heating rods 6.Cultivation cup 5 is placed in to cleansing bath tub 3, and each cleansing bath tub 3 is placed six and is cultivated cup 5.Aeration pump 8 is placed in to Water-accumulating water cylinder 4.Thermometer 7 is placed in to cleansing bath tub 3 and Water-accumulating water cylinder 4, one, every cylinder.Fluorescent lamp 9 is placed in to 2 ~ 3 meters of support 1 oblique uppers, and parallel with support 1.Fluorescent lamp 9 links with time controller 10, with the controlled light time.
According to above-mentioned size and method, to build Daphnia magna culture apparatus and carry out Daphnia magna cultivation, cultural method comprises the steps:
(1) pour appropriate running water into cleansing bath tub and Water-accumulating water cylinder, aeration pump is opened, Water-accumulating water cylinder running water is carried out to continuous aeration, to remove chlorine residue in running water, utilize heating rod and thermometer that water temperature in cleansing bath tub and Water-accumulating water cylinder is adjusted to 20 ± 1 ℃ simultaneously.
(2) water in three days above Water-accumulating water cylinders of aeration is added to and is cultivated in cup, every glass of 2000ml, in cultivating cup, inoculate purebred, firm the be born Daphnia magna children flea of 6 ~ 24h, each cultivates 70 to 80 of cup inoculations, inoculation number is difficult for too much, under this density, and Daphnia magna fast growth, breeding amount is large, simultaneously the record inoculation date.
(3) chlorella vulgaris that does not contain medium is diluted in and is cultivated in cup, (chlorella vulgaris density is about 1 * 10 to make cultivation cup present light green
6~ 4 * 10
6individual/ml).Recommendation chlorella vulgaris of the present invention carrys out feeding Daphnia magna, and its reason is that chlorella vulgaris particle diameter is less, and suspension is higher, precipitates lessly, can be distributed in more uniformly and cultivate in cup.Make on the one hand Daphnia magna all can filter food food everywhere at cultivation cup, can prevent on the other hand bottom algae liquid excessive concentration, be attached on Daphnia magna feeler, affect Daphnia magna proper motion, thereby affect its life state and fertility.
(4) in distance is cultivated the scope of 2 ~ 3 meters, cup, adjusting the position of fluorescent lamp, is that 800 ~ 1000lux(intensity of illumination deficiency can increase fluorescent lamp quantity if make to cultivate a cup place intensity of illumination), during Daphnia magna is dispersed in and cultivates glass simultaneously.Prevent that the local density that the phototaxis due to Daphnia magna causes is excessive, affect Daphnia magna parthenogenesis.With time controller, control illumination every day 16h, dark 8h.
(5) between culture period, within every 3 days, change and cultivate the medium-and-large-sized flea culture fluid of cup, each 500ml that changes, the water that changes is in Water-accumulating water cylinder, and aeration is more than three days, and temperature is the water of 20 ± 1 ℃, prevent that environment cataclysm from affecting Daphnia magna animation, remove simultaneously and be deposited on Daphnia magna shelling, algae group and the dead Daphnia magna of cultivating cup bottom, append afterwards food, make to cultivate cup and present light green.
(6), while cultivating to 7 days left and right, Daphnia magna will produce the 1st tire children flea (the female flea of the 7th age in days produces young flea), the separated female flea of screen cloth and the young flea that with upper and lower sleeve, are combined, and upper strata screen cloth is 30 orders, lower screen is 60 orders.Now produce the general quantity of young flea few, small, unstable, abandon it.Female flea is put back to former cultivation cup and continues to cultivate.
(7) after this approximately within every 3 days, just having a new tire children flea produces.The female flea of recommendations for selection 16 ~ 22 ages in days of the present invention produces young flea and carries out Daphnia magna toxicity test.Therefore, eliminate 16 ages in days before female flea produce young flea, collect female 6 ~ 24h that flea the produces children of 16 ~ 22 ages in days fleas and test or be inoculated into another one and cultivate continuation cultivation in cup, and record date.
(8) treat that Daphnia magna surpasses 22 ages in days and can eliminate.
Can obtain according to the method described above cultivating Daphnia magnas more than 3 generations, and be all the female fleas of 16 ~ 22 ages in days and produce young flea, their quantity is large, evenly individual, fast growth, and fertility is strong, and life state is stable, is suitable for very much Daphnia magna toxotest and tests.In addition, because Daphnia magna quantity is large, can be in water treatment field, remove in water the tests such as zooplankton enough test materials are provided.
Claims (10)
1. Daphnia magna laboratory fast culture device, is characterized in that described culture apparatus comprises support, base plate, cleansing bath tub, Water-accumulating water cylinder, cultivation cup, heating rod, thermometer, aeration pump, fluorescent lamp, time controller; The bottom of described support is provided with base plate, is placed with Water-accumulating water cylinder and cleansing bath tub on base plate, is provided with thermometer, aeration pump and heating rod in Water-accumulating water cylinder, and cleansing bath tub is provided with thermometer, heating rod and cultivation cup; Described fluorescent lamp is positioned at support oblique upper parallel with support, and time controller is connected with fluorescent lamp.
2. Daphnia magna according to claim 1 laboratory fast culture device, is characterized in that described support is that angle iron builds, and is divided into three layers, and every layer height is adjustable, and the bottom is placed a Water-accumulating water cylinder, places two cleansing bath tubs for upper two-layer every layer.
3. Daphnia magna according to claim 1 laboratory fast culture device, is characterized in that described base plate is wooden or glass system.
4. Daphnia magna according to claim 1 and 2 laboratory fast culture device, is characterized in that described cleansing bath tub and Water-accumulating water cylinder are glass system square, and cultivating cup is the round glassware of splendid attire 2000ml water.
5. Daphnia magna according to claim 1 laboratory fast culture device, is characterized in that described heating rod is the family expenses heating rods of breeding fish.
6. utilize Daphnia magna laboratory fast culture device described in the arbitrary right of claim 1-5 to carry out a method for Daphnia magna laboratory fast culture, it is characterized in that described cultural method step is as follows:
(1) running water is poured into cleansing bath tub and Water-accumulating water cylinder, aeration pump is opened, Water-accumulating water cylinder running water is carried out to continuous aeration, to remove chlorine residue in running water, utilize heating rod and thermometer that water temperature in cleansing bath tub and Water-accumulating water cylinder is adjusted to 20 ± 1 ℃ simultaneously;
(2) water in the above Water-accumulating water cylinder of aeration three days is added to and is cultivated in cup, every glass of 2000ml inoculates purebred, firm the be born Daphnia magna children flea of 6 ~ 24h in cultivating cup, and each cultivates 70 to 80 of cup inoculations, simultaneously the record inoculation date;
(3) chlorella vulgaris that does not contain medium is diluted in and is cultivated in cup, make to cultivate cup and present light green;
(4) adjust the position of fluorescent lamp, making to cultivate cup place intensity of illumination is 800 ~ 1000lux;
(5) between culture period, within every 3 days, change and cultivate the medium-and-large-sized flea culture fluid of cup, each 500ml that changes, the water that changes is in Water-accumulating water cylinder, aeration is more than three days, and temperature is the water of 20 ± 1 ℃, removes simultaneously and is deposited on Daphnia magna shelling, algae group and the dead Daphnia magna of cultivating cup bottom, append afterwards food, make to cultivate cup and present light green;
(6) when Daphnia magna produces young flea, with the separated female flea of screen cloth and young flea, before superseded 16 ages in days, female flea produces young flea, and female flea is put back in former cultivation cup and continues to cultivate;
(7) collect 16 ~ 22 ages in days mother 6 ~ 24h that flea produces children fleas and test or be inoculated into continuation cultivation in another one cultivation cup, and record date;
(8) treat that Daphnia magna surpasses 22 ages in days and can eliminate.
7. Daphnia magna according to claim 6 laboratory fast culture process, is characterized in that, in described step (3), chlorella vulgaris density is 1 * 10
6-4 * 10
6individual/ml.
8. Daphnia magna according to claim 6 laboratory fast culture process, is characterized in that in described step (4), controls illumination every day 16h, dark 8h.
9. Daphnia magna according to claim 6 laboratory fast culture process, is characterized in that, in described step (6), screen cloth is the screen cloth that upper and lower sleeve is combined.
10. Daphnia magna according to claim 9 laboratory fast culture process, is characterized in that described upper strata screen cloth is 30 orders, and lower screen is 60 orders.
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| CN104273101A (en) * | 2014-10-24 | 2015-01-14 | 南京大学 | Artificial cultivation method of Moina micrura Kurz or Ceriodaphnia cornuta Sars |
| CN105918260A (en) * | 2016-07-13 | 2016-09-07 | 环境保护部南京环境科学研究所 | Running water type exposure device applied to small aquatic organisms |
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| CN107094721A (en) * | 2017-07-10 | 2017-08-29 | 佛山市水创科联生态科技有限公司 | The primary Daphnia magna enriching apparatus in field and enrichment method |
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