CN103598150B - Daphnia magna laboratory fast culture device and cultural method - Google Patents
Daphnia magna laboratory fast culture device and cultural method Download PDFInfo
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- CN103598150B CN103598150B CN201310624588.3A CN201310624588A CN103598150B CN 103598150 B CN103598150 B CN 103598150B CN 201310624588 A CN201310624588 A CN 201310624588A CN 103598150 B CN103598150 B CN 103598150B
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- 241001494246 Daphnia magna Species 0.000 title claims abstract description 83
- 238000000034 method Methods 0.000 title claims abstract description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 72
- 238000010438 heat treatment Methods 0.000 claims abstract description 26
- 238000005273 aeration Methods 0.000 claims abstract description 24
- 238000012360 testing method Methods 0.000 claims abstract description 16
- 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
- 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
- 231100000820 toxicity test Toxicity 0.000 abstract description 4
- 230000035558 fertility Effects 0.000 description 4
- 230000008186 parthenogenesis Effects 0.000 description 4
- 238000009395 breeding Methods 0.000 description 3
- 230000001488 breeding effect Effects 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
- 239000002244 precipitate Substances 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
- 238000012423 maintenance Methods 0.000 description 1
- 238000004519 manufacturing process Methods 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
- 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
Daphnia magna laboratory fast culture device and cultural method, belong to zooplankton and cultivate field.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, base plate is placed with Water-accumulating water cylinder and cleansing bath tub, 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 and parallel with support, and time controller is connected with fluorescent lamp.The present invention is simple and easy to do, workable, and culture device device is easily built, without the need to loaded down with trivial details device structure; The Daphnia magna reproduction speed of turning out is fast, and quantity is large, and life state is stablized, individual difference is little, flea class toxicity test result is stablized, reliably, test mass is ensured, can be in water treatment field simultaneously, removes the tests such as zooplankton in water pollutes and provides enough test materials.
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 (Daphniamagna) belongs to Arthropoda, Crustachia, cladocera, is a kind of common freshwater zooplankton, in aquatic ecosystem, has critical role.Because it has widely distributed, life cycle is short, and reproduction speed is fast, and reproduction amount is high, is easy to cultivate, and is widely used in Aquatic Toxicity Test to features such as poisonous substance sensitivities.
Current Daphnia magna is internationally recognized code test indicator organism, and many countries have also established the standard method of Daphnia magna toxotest simultaneously, has done relevant regulation to the condition of culture of test Daphnia magna.But the Daphnia magna in the most of laboratory of China is cultivated and lacked effective condition of culture or device at present, be difficult to ensure culture effect, this makes the culture environment of Daphnia magna stable not, and Daphnia magna is difficult to keep parthenogenesis for a long time, and Daphnia magna quantity is difficult to ensure.Therefore be difficult to obtain the Daphnia magna needed for test, such as Daphnia magna is different for the length of time, and 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 the defect overcoming existing Daphnia magna culture technique, a kind of laboratory Daphnia magna culture apparatus of improvement is provided and utilizes this Daphnia magna culture apparatus to carry out the method for Daphnia magna cultivation.This device can under manual control condition, realize the breeding of Daphnia magna immediate stability, and be in parthenogenesis state for a long time, individual straight uniform, life state is active, the quality of Daphnia magna toxicity test can be ensured, and more simple than standard method, meet the requirement of laboratory fast culture and use.
Daphnia magna laboratory of the present invention 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, base plate is placed with Water-accumulating water cylinder and cleansing bath tub, 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 and parallel with support, and time controller is connected with fluorescent lamp.
In said apparatus, described support is that angle iron builds, and be divided into three layers, every layer height is adjustable, and a Water-accumulating water cylinder placed by the bottom, 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, and this base plate can be wooden also can be glass system, ensures to have enough bearing capacities.
In said apparatus, described cleansing bath tub is glass system, square, is fixed in cleansing bath tub at cleansing bath tub sidewall near bottom sucker by heating rod, each cleansing bath tub be provided with a thermometer, two heating rods, six cultivate cups.
In said apparatus, described Water-accumulating water cylinder is glass system, square, and be fixed on Water-accumulating water cylinder near bottom sucker by heating rod at Water-accumulating water cylinder sidewall, each Water-accumulating water cylinder is provided with a thermometer, aeration pump, two heating rods.
In said apparatus, described cultivation cup is can the round glassware of splendid attire 2000ml water, one-shot forming non-cohesive material.
In said apparatus, described heating rod is roughly to regulate the family expenses of temperature to breed fish heating rod.
In said apparatus, described thermometer instruction 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 described, fluorescent lamp distance bracket oblique upper, 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) pour running water into cleansing bath tub and Water-accumulating water cylinder, aeration pump is opened, continuous aeration is carried out to Water-accumulating water cylinder running water, to remove chlorine residue in running water, utilize heating rod and thermometer by lower water temperature in cleansing bath tub and Water-accumulating water cylinder to 20 ± 1 DEG C simultaneously;
(2) water in the aeration Water-accumulating water cylinder of more than three days be added to cultivate in cup, every glass of 2000ml, cultivate in cup inoculate purebred, to be just born the Daphnia magna children flea of 6 ~ 24h, each cultivation cup inoculation 70 to 80, simultaneously record inoculation date;
(3) chlorella vulgaris not containing medium is diluted in cultivation cup, makes to cultivate cup and present light green;
(4) adjust the position of fluorescent lamp, make cultivation cup place intensity of illumination be 800 ~ 1000lux;
(5) between culture period, within every 3 days, change and cultivate the medium-and-large-sized flea culture fluid of cup, each replacing 500ml, the water that changes is in Water-accumulating water cylinder, aeration more than three days, temperature is the water of 20 ± 1 DEG C, and the Daphnia magna be deposited on bottom cultivation cup of removing simultaneously shells, algae is rolled into a ball and dead Daphnia magna, add food afterwards, make to cultivate cup and present light green;
(6) when Daphnia magna produces young flea, be separated female flea and young flea with the screen cloth in suitable aperture, the father's former wife flea of eliminating 16 ages in days produces young flea, and female flea is put back to former cultivation cup and continues to cultivate;
(7) collect 16 ~ 22 ages in days female 6 ~ 24h that flea produces children flea to carry out testing or being inoculated in another one cultivation cup continuing to cultivate, and record date;
(8) treat that Daphnia magna can be eliminated more than 22 ages in days.
Can obtain the Daphnia magna in cultivation 3 generation more than according to the method described above, 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 stable.
The present invention has following beneficial effect:
(1) Daphnia magna laboratory provided by the invention fast culture apparatus structure simple, to manufacture and easy to maintenance, expense is few, and material source is wide, easily builds.And can Appropriate application laboratory solid space, reduce floor space.
(2) Daphnia magna laboratory provided by the invention fast culture process can be cultivated in room conditions all the year round, and Daphnia magna breeding amount is large, fast growth, for Daphnia magna toxotest provides sufficient at any time, and can the test material of guarantee test quality.In addition, because Daphnia magna quantity is large, can be in water treatment field, in removal water, zooplankton etc. be tested and provide enough test materials.
(3) Daphnia magna laboratory provided by the invention fast culture process is without the need to frequently changing water, and each quantity of exchanged water is few, greatly reduces workload.
(4) Daphnia magna laboratory provided by the invention fast culture process selects chlorella vulgaris to be Daphnia magna food, because chlorella vulgaris suspension is good, not easily precipitates, and food can be made to be dispersed in Daphnia magna and cultivate in cup, is beneficial to Daphnia magna filter food.Green alga can be avoided to be attached on Daphnia magna feeler simultaneously, thus affect Daphnia magna locomitivity, the unfavorable problem such as life state and fertility.
(5) this cultural method can make Daphnia magna photoenvironment stablize, and prevents Daphnia magna local density excessive, and make to cultivate female flea in cup density stabilized, the moment keeps parthenogenesis state.
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 amendment not having creative contribution as required after reading this specification, as long as but be all subject to the protection of Patent Law in right of the present invention.
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.
The title of this each main member of device, material and specification as follows:
Support 1: iron, long 120cm, wide 40cm, high 150cm.Be divided into three layers, the superiors 30cm, intermediate layer and orlop 55cm.
Base plate 2: wooden, also can glass system, has 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, circular, diameter 14cm, high 20cm, also can replace with 2000ml beaker.
Heating rod 6:100w, temperature is adjustable 18-32 DEG C.
Thermometer 7: 20 ± 1 DEG C can be surveyed.
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 places a Water-accumulating water cylinder 4.Heating rod 6 is placed in cleansing bath tub 3 and Water-accumulating water cylinder 4 sidewall near bottom position, each heating rod 6 two suckers are fixed, and each cleansing bath tub 3 and Water-accumulating water cylinder 4 place two heating rods 6.Cultivation cup 5 is placed in cleansing bath tub 3, and each cleansing bath tub 3 is placed six and is cultivated cup 5.Aeration pump 8 is placed in Water-accumulating water cylinder 4.Thermometer 7 is placed in cleansing bath tub 3 and Water-accumulating water cylinder 4, one, every cylinder.Fluorescent lamp 9 is placed in 2 ~ 3 meters, support 1 oblique upper, 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, 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, continuous aeration is carried out to Water-accumulating water cylinder running water, to remove chlorine residue in running water, utilize heating rod and thermometer by lower water temperature in cleansing bath tub and Water-accumulating water cylinder to 20 ± 1 DEG C simultaneously.
(2) water in the aeration Water-accumulating water cylinder of more than three days is added in cultivation cup, every glass of 2000ml, cultivate in cup inoculate purebred, to be just born the Daphnia magna children flea of 6 ~ 24h, each cultivation cup inoculation 70 to 80, inoculation number is not easily too much, under this density, and Daphnia magna fast growth, breeding amount is large, simultaneously the record inoculation date.
(3) be diluted in cultivation cup by the chlorella vulgaris not containing medium, (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 less, can be distributed in more uniformly and cultivate in cup.Make Daphnia magna all can filter food food everywhere at cultivation cup on the one hand, bottom algae liquid excessive concentration can be prevented on the other hand, be attached on Daphnia magna feeler, affect Daphnia magna proper motion, thus affect its life state and fertility.
(4) in the scope that distance cultivates 2 ~ 3 meters, cup, the position of fluorescent lamp is adjusted, if make cultivation cup place intensity of illumination be that 800 ~ 1000lux(intensity of illumination deficiency can increase fluorescent lamp quantity), Daphnia magna is dispersed in and cultivates in cup simultaneously.Prevent the local density that causes due to the phototaxis of Daphnia magna 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 replacing 500ml, the water that changes is in Water-accumulating water cylinder, aeration more than three days, and temperature is the water of 20 ± 1 DEG C, prevent environment cataclysm from affecting Daphnia magna animation, the Daphnia magna be deposited on bottom cultivation cup of removing simultaneously shells, algae is rolled into a ball and dead Daphnia magna, adds food afterwards, makes cultivation cup present light green.
(6), when cultivating by about 7 days, Daphnia magna will produce the 1st tire children flea (namely the female flea of the 7th age in days produces young flea), and be separated female flea and young flea with the screen cloth that upper and lower sleeve is combined, upper strata screen cloth is 30 orders, and lower screen is 60 orders.Now the general quantity of produced young flea is few, small, unstable, abandons it.Female flea is put back to former cultivation cup and continues to cultivate.
(7) within after this approximately every 3 days, just have a new tire children flea to produce.The female flea of recommendations for selection 16 ~ 22 age in days of the present invention produces young flea and carries out Daphnia magna toxicity test.Therefore, the father's former wife flea of eliminating 16 ages in days produces young flea, collects 16 ~ 22 ages in days female 6 ~ 24h that flea produces children flea and carries out testing or being inoculated in another one cultivation cup continuing to cultivate, and record date.
(8) treat that Daphnia magna can be eliminated more than 22 ages in days.
Can obtain the Daphnia magna in cultivation 3 generation more than according to the method described above, 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 stablized, and is suitable for very much Daphnia magna toxotest and tests.In addition, because Daphnia magna quantity is large, can be in water treatment field, in removal water, zooplankton etc. be tested and provide enough test materials.
Claims (8)
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; Described support is that angle iron builds, be divided into three layers, every layer height is adjustable, bottom is provided with base plate, and the base plate of the bottom places a Water-accumulating water cylinder, and upper two-layer base plate places two cleansing bath tubs for every layer, thermometer, aeration pump and heating rod is provided with in Water-accumulating water cylinder, cleansing bath tub is provided with thermometer, heating rod and cultivation cup, and it is square that described cleansing bath tub and Water-accumulating water cylinder are glass system, cultivates the circular glass goods that cup is splendid attire 2000ml water; Described fluorescent lamp is positioned at support oblique upper and parallel with support, and time controller is connected with fluorescent lamp.
2. Daphnia magna laboratory according to claim 1 fast culture device, is characterized in that described base plate is wooden or glass system.
3. Daphnia magna laboratory according to claim 1 fast culture device, is characterized in that described heating rod is that family expenses are breeded fish heating rod.
4. utilize the Daphnia magna laboratory fast culture device described in the arbitrary claim of claim 1-3 to carry out a method for Daphnia magna laboratory fast culture, it is characterized in that described cultural method step is as follows:
(1) pour running water into cleansing bath tub and Water-accumulating water cylinder, aeration pump is opened, continuous aeration is carried out to Water-accumulating water cylinder running water, to remove chlorine residue in running water, utilize heating rod and thermometer by lower water temperature in cleansing bath tub and Water-accumulating water cylinder to 20 ± 1 DEG C simultaneously;
(2) water in the aeration Water-accumulating water cylinder of more than three days be added to cultivate in cup, every glass of 2000ml, cultivate in cup inoculate purebred, to be just born the Daphnia magna children flea of 6 ~ 24h, each cultivation cup inoculation 70 to 80, simultaneously record inoculation date;
(3) chlorella vulgaris not containing medium is diluted in cultivation cup, makes to cultivate cup and present light green;
(4) adjust the position of fluorescent lamp, make cultivation cup place intensity of illumination be 800 ~ 1000lux;
(5) between culture period, within every 3 days, change and cultivate the medium-and-large-sized flea culture fluid of cup, each replacing 500ml, the water that changes is in Water-accumulating water cylinder, aeration more than three days, temperature is the water of 20 ± 1 DEG C, and the Daphnia magna be deposited on bottom cultivation cup of removing simultaneously shells, algae is rolled into a ball and dead Daphnia magna, add food afterwards, make to cultivate cup and present light green;
(6) when Daphnia magna produces young flea, be separated female flea and young flea with screen cloth, the father's former wife flea of eliminating 16 ages in days produces young flea, and female flea is put back in former cultivation cup and continues to cultivate;
(7) collect 16 ~ 22 ages in days female 6 ~ 24h that flea produces children flea to carry out testing or being inoculated in another one cultivation cup continuing to cultivate, and record date;
(8) treat that Daphnia magna can be eliminated more than 22 ages in days.
5. Daphnia magna laboratory according to claim 4 fast culture process, is characterized in that, in described step (3), chlorella vulgaris density is 1 × 10
6-4 × 10
6individual/ml.
6. Daphnia magna laboratory according to claim 4 fast culture process, is characterized in that in described step (4), controls illumination every day 16h, dark 8h.
7. Daphnia magna laboratory according to claim 4 fast culture process, is characterized in that, in described step (6), screen cloth is the screen cloth that upper and lower sleeve is combined.
8. Daphnia magna laboratory according to claim 7 fast culture process, it is characterized in that upper strata screen cloth is 30 orders, lower screen is 60 orders.
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| CN104222027B (en) * | 2014-10-13 | 2016-06-15 | 长江流域水环境监测中心 | The one highly enriched food of kind and preparation method |
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| CN107094721B (en) * | 2017-07-10 | 2021-01-26 | 广东民信科技发展有限公司 | Wild large native daphnia magna enrichment device and enrichment method |
| CN107897127A (en) * | 2017-11-21 | 2018-04-13 | 佛山科学技术学院 | A kind of cultural method of purebred Daphnia magna for water ecology reparation |
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