CN110235862A - A method of utilizing the toxicity of Drosophila melanogaster behavior stress reaction evaluation pollutant - Google Patents
A method of utilizing the toxicity of Drosophila melanogaster behavior stress reaction evaluation pollutant Download PDFInfo
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- CN110235862A CN110235862A CN201910533336.7A CN201910533336A CN110235862A CN 110235862 A CN110235862 A CN 110235862A CN 201910533336 A CN201910533336 A CN 201910533336A CN 110235862 A CN110235862 A CN 110235862A
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- 241000255601 Drosophila melanogaster Species 0.000 title claims abstract description 42
- 239000003344 environmental pollutant Substances 0.000 title claims abstract description 38
- 231100000719 pollutant Toxicity 0.000 title claims abstract description 38
- 238000000034 method Methods 0.000 title claims abstract description 20
- 231100000419 toxicity Toxicity 0.000 title claims abstract description 20
- 230000001988 toxicity Effects 0.000 title claims abstract description 20
- 238000011156 evaluation Methods 0.000 title claims abstract description 8
- 239000007787 solid Substances 0.000 claims abstract description 28
- 238000001514 detection method Methods 0.000 claims abstract description 9
- XEFQLINVKFYRCS-UHFFFAOYSA-N Triclosan Chemical compound OC1=CC(Cl)=CC=C1OC1=CC=C(Cl)C=C1Cl XEFQLINVKFYRCS-UHFFFAOYSA-N 0.000 claims description 21
- HBGGXOJOCNVPFY-UHFFFAOYSA-N diisononyl phthalate Chemical group CC(C)CCCCCCOC(=O)C1=CC=CC=C1C(=O)OCCCCCCC(C)C HBGGXOJOCNVPFY-UHFFFAOYSA-N 0.000 claims description 21
- 229960003500 triclosan Drugs 0.000 claims description 21
- CHHHXKFHOYLYRE-UHFFFAOYSA-M 2,4-Hexadienoic acid, potassium salt (1:1), (2E,4E)- Chemical compound [K+].CC=CC=CC([O-])=O CHHHXKFHOYLYRE-UHFFFAOYSA-M 0.000 claims description 8
- 229920001817 Agar Polymers 0.000 claims description 8
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 8
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 8
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 8
- 229930006000 Sucrose Natural products 0.000 claims description 8
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 8
- 240000008042 Zea mays Species 0.000 claims description 8
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 claims description 8
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 8
- 239000008272 agar Substances 0.000 claims description 8
- 239000001110 calcium chloride Substances 0.000 claims description 8
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 8
- 235000013312 flour Nutrition 0.000 claims description 8
- 239000008103 glucose Substances 0.000 claims description 8
- 235000009973 maize Nutrition 0.000 claims description 8
- 239000004302 potassium sorbate Substances 0.000 claims description 8
- 235000010241 potassium sorbate Nutrition 0.000 claims description 8
- 229940069338 potassium sorbate Drugs 0.000 claims description 8
- 239000002904 solvent Substances 0.000 claims description 8
- 239000005720 sucrose Substances 0.000 claims description 8
- QFOHBWFCKVYLES-UHFFFAOYSA-N Butylparaben Chemical compound CCCCOC(=O)C1=CC=C(O)C=C1 QFOHBWFCKVYLES-UHFFFAOYSA-N 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 231100000331 toxic Toxicity 0.000 claims description 2
- 230000002588 toxic effect Effects 0.000 claims description 2
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims 1
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 239000000460 chlorine Substances 0.000 claims 1
- 229910052801 chlorine Inorganic materials 0.000 claims 1
- DROMNWUQASBTFM-UHFFFAOYSA-N dinonyl benzene-1,2-dicarboxylate Chemical group CCCCCCCCCOC(=O)C1=CC=CC=C1C(=O)OCCCCCCCCC DROMNWUQASBTFM-UHFFFAOYSA-N 0.000 claims 1
- 238000011160 research Methods 0.000 abstract description 3
- 239000002609 medium Substances 0.000 description 22
- 241000255581 Drosophila <fruit fly, genus> Species 0.000 description 16
- 230000009194 climbing Effects 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 5
- 239000003153 chemical reaction reagent Substances 0.000 description 4
- 238000011109 contamination Methods 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- 210000001015 abdomen Anatomy 0.000 description 3
- 229960000935 dehydrated alcohol Drugs 0.000 description 3
- 229960001760 dimethyl sulfoxide Drugs 0.000 description 3
- 238000005286 illumination Methods 0.000 description 3
- 241000255925 Diptera Species 0.000 description 2
- 206010020591 Hypercapnia Diseases 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 235000013399 edible fruits Nutrition 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000009533 lab test Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- XNGIFLGASWRNHJ-UHFFFAOYSA-N phthalic acid Chemical compound OC(=O)C1=CC=CC=C1C(O)=O XNGIFLGASWRNHJ-UHFFFAOYSA-N 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 239000011550 stock solution Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 231100000048 toxicity data Toxicity 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- IWPHJSSEOIHZRP-UHFFFAOYSA-N benzoic acid;butyl acetate Chemical compound CCCCOC(C)=O.OC(=O)C1=CC=CC=C1 IWPHJSSEOIHZRP-UHFFFAOYSA-N 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 229960003742 phenol Drugs 0.000 description 1
- DCKVNWZUADLDEH-UHFFFAOYSA-N sec-butyl acetate Chemical compound CCC(C)OC(C)=O DCKVNWZUADLDEH-UHFFFAOYSA-N 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000012549 training Methods 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/111—Aromatic compounds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
- A23K20/24—Compounds of alkaline earth metals, e.g. magnesium
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/90—Feeding-stuffs specially adapted for particular animals for insects, e.g. bees or silkworms
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Engineering & Computer Science (AREA)
- Animal Husbandry (AREA)
- Zoology (AREA)
- Food Science & Technology (AREA)
- Microbiology (AREA)
- Health & Medical Sciences (AREA)
- Environmental Sciences (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Physiology (AREA)
- Biomedical Technology (AREA)
- Botany (AREA)
- Mycology (AREA)
- Insects & Arthropods (AREA)
- Birds (AREA)
- Animal Behavior & Ethology (AREA)
- Biodiversity & Conservation Biology (AREA)
- Biochemistry (AREA)
- Inorganic Chemistry (AREA)
- Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The present invention provides a kind of methods of toxicity using Drosophila melanogaster behavior stress reaction evaluation pollutant, belong to toxicity research technical field, the following steps are included: 1) Drosophila melanogaster is placed in 1~15d of culture on the solid medium containing pollutant, the quantity that detection Drosophila melanogaster is creeped in 10s more than 10cm, is obtained line rate；2) Drosophila melanogaster is placed in 1~15d of culture on solid medium, line rate is obtained in the quantity that detection Drosophila melanogaster is creeped in 10s more than 10cm；3) when significant difference occurs in the line rate excessively that line rate and step 2) excessively that the step 1) obtains obtain, show that pollutant has toxicity.The toxicity of pollutant can be accurately evaluated using method provided by the invention, provide the support of theoretical and experimental data for the dosage and concentration of the pollutant of agriculturally reasonable employment.
Description
Technical field
The invention belongs to toxicity research technical fields more particularly to a kind of utilization Drosophila melanogaster behavior stress reaction to evaluate dirt
Contaminate the method for the toxicity of object.
Background technique
The compound amounts that U.S. chemical abstract society (CAS) includes at present are considerably beyond U.S. national library of medicine toxicity
The toxicity data recorded in Data web site (TOXNET) is learned, a large amount of existing substances do not have relevant toxicity data in environment, and
And be greatly present in our daily life, it is close with human contact.If these are largely without the object of toxicity assessment
Matter is for a long time and human contact, it will causes grave danger safely to social publilc health.
The research of toxic effect is concentrated mainly on plant, aquatic animal and mammal at present, grinds relative to terrestrial insect
Study carefully less.
Summary of the invention
In view of this, it is an object of the invention to a kind of toxicity using Drosophila melanogaster behavior stress reaction evaluation pollutant
Method, the toxicity of pollutant can be evaluated, accurately using method provided by the invention for the pollution of agriculturally reasonable employment
The dosage and concentration of object provide theoretical and experimental data and support.
In order to achieve the above-mentioned object of the invention, the present invention provides following technical schemes:
The present invention provides a kind of methods of toxicity using Drosophila melanogaster evaluation pollutant, comprising the following steps:
1) Drosophila melanogaster is placed in 1~15d of culture on the solid medium containing pollutant, detects Drosophila melanogaster in 10s
Line rate is obtained in the quantity inside creeped more than 10cm；
2) Drosophila melanogaster is placed in 1~15d of culture on solid medium, detection Drosophila melanogaster is creeped in 10s is more than
Line rate is obtained in the quantity of 10cm；
3) when significant difference occurs in the line rate excessively that line rate and step 2) excessively that the step 1) obtains obtain, show to pollute
Object has toxicity.
Preferably, the pollutant includes triclosan and/or diisononyl phthalate.
Preferably, when the pollutant is triclosan, triclosan is dense in the solid medium containing triclosan
Degree is 10~500mg/L.
Preferably, described to contain diisononyl phthalate when the pollutant is diisononyl phthalate
Solid medium in diisononyl phthalate volumetric concentration be 0.1~1%.
Preferably, the solid medium containing pollutant takes water as a solvent, and every liter further include: 70~80g maize flour,
30~35g yeast, 8~12g agar, 0.5~1g CaCl2, 30~33g sucrose, 60~65g glucose, 1~3g potassium sorbate
With 5% butyl p-hydroxybenzoate solution of 15ml.
Preferably, the solid medium takes water as a solvent, and every liter includes: 70~80g maize flour, 30~35g yeast, 8
~12g agar, 0.5~1g CaCl2, 30~33g sucrose, 60~65g glucose, 1~3g potassium sorbate and 15ml 5% be to hydroxyl
Yl benzoic acid butyl acetate solution.
Preferably, the female fly of the Drosophila melanogaster and male fly are individually cultivated.
Preferably, it is 24~26 DEG C that the feeding condition, which includes: the temperature of the culture, and the Light To Dark Ratio of the culture is
12h:12h；The ambient humidity of the culture is 50~60%.
The present invention provides a kind of methods of toxicity using Drosophila melanogaster behavior stress reaction evaluation pollutant, including with
Lower step: 1) being placed in 1~15d of culture on the solid medium containing pollutant for Drosophila melanogaster, detects Drosophila melanogaster in 10s
Creep be more than 10cm quantity, line rate is obtained；2) Drosophila melanogaster is placed in 1~15d of culture on solid medium, detects black abdomen
Line rate is obtained in the quantity that drosophila is creeped in 10s more than 10cm；3) when the line rate of crossing that the step 1) obtains is obtained with step 2)
To cross line rate there is significant difference when, show pollutant have toxicity.It can accurately be commented using method provided by the invention
The toxicity of valence pollutant provides the support of theoretical and experimental data for the dosage and concentration of the pollutant of agriculturally reasonable employment.
Detailed description of the invention
Fig. 1 is that female fly climbed line rate after being exposed to triclosan；
Fig. 2 is that male fly climbed line rate after being exposed to triclosan；
Fig. 3 is that female fly climbed line rate after being exposed to diisononyl phthalate；
Fig. 4 is that male fly climbed line rate after being exposed to diisononyl phthalate；
Specific embodiment
The present invention provides a kind of methods of toxicity using Drosophila melanogaster behavior stress reaction evaluation pollutant, including with
Lower step: 1) being placed in 1~15d of culture on the solid medium containing pollutant for Drosophila melanogaster, detects Drosophila melanogaster in 10s
Creep be more than 10cm quantity, line rate is obtained；2) Drosophila melanogaster is placed in 1~15d of culture on solid medium, detects black abdomen
Line rate is obtained in the quantity that drosophila is creeped in 10s more than 10cm；3) when the line rate of crossing that the step 1) obtains is obtained with step 2)
To cross line rate there is significant difference when, show pollutant have toxicity.
Drosophila melanogaster is placed in 1~15d of culture on the solid medium containing pollutant by the present invention, and detection Drosophila melanogaster exists
Line rate is obtained in the quantity creeped in 10s more than 10cm.
In the present invention, the Drosophila melanogaster preferably selects w118Wild type Drosophila melanogaster, the present invention is to the w118It is wild
The source of type Drosophila melanogaster is not particularly limited, using conventional commercial.
In the present invention, the female fly of the Drosophila melanogaster and male fly are preferably individually cultivated.In the present invention, the female fly and
The quantity of male fly is independently 30；The female fly and male fly, which preferably hatch in 8h, to be obtained.
In the present invention, the solid medium containing pollutant preferably takes water as a solvent, and every liter further preferably includes: 70
~80g maize flour, 30~35g yeast, 8~12g agar, 0.5~1g CaCl2, 30~33g sucrose, 60~65g glucose, 1
5% butyl p-hydroxybenzoate solution of~3g potassium sorbate and 15ml；More preferably include 77.7g maize flour, 32.19g yeast,
10.6g agar, 0.726g CaCl2, 31.62g sucrose, 5% para hydroxybenzene first of 63.2g glucose, 2g potassium sorbate and 15ml
Acid butyl ester solution.
In the present invention, the condition of the culture preferably includes: the temperature of the culture is preferably 24~26 DEG C, more preferably
It is 25 DEG C；The Light To Dark Ratio of the culture is 12h:12h；The ambient humidity of the culture is 50~60%.The present invention is to the training
Required intensity of illumination is not particularly limited when supporting, and the illumination using those skilled in the art's conventional illumination culture Drosophila melanogaster is strong
Degree.Present invention preferably employs constant incubators to cultivate Drosophila melanogaster.
In the present invention, the pollutant preferably includes triclosan and/or diisononyl phthalate.In the present invention
In, when the pollutant is preferably triclosan, the concentration of triclosan is preferably in the solid medium containing triclosan
10~500mg/L, specially 10,100,250 and 500mg/L, the triclosan are preferably bought in Shanghai Mike's woods biochemical technology
Co., Ltd, purity 97%.The triclosan is preferably dissolved in dimethyl sulfoxide by the present invention, is added in culture medium, is matched
Corresponding concentration is made.
In the present invention, described to contain phthalic acid when the pollutant is preferably diisononyl phthalate
The volumetric concentration of diisononyl phthalate is preferably 0.1~1% in the solid medium of dinonyl, specially 0.1,
0.2,0.5 and 1%, the diisononyl phthalate is preferably bought in Aladdin chemical reagents corporation, purity 99.5%.
The diisononyl phthalate is preferably dissolved in dehydrated alcohol by the present invention, is then added in culture medium, is configured to corresponding
Concentration.
In the present invention, when the pollutant is triclosan, the time of the Drosophila melanogaster culture is preferably 1~15d,
Specially 1d, 3d, 5d, 7d, 9d, 11d, 13d and 15d；When the pollutant is diisononyl phthalate, the black abdomen
The time of drosophila culture is preferably 1~15d, specially 1d, 5d, 10d and 15d.
Drosophila melanogaster is placed in 1~15d of culture on solid medium by the present invention, and detection Drosophila melanogaster is creeped super in 10s
Line rate is obtained in the quantity for crossing 10cm.
In the present invention, the solid medium preferably takes water as a solvent, every liter include: 70~80g maize flour, 30~
35g yeast, 8~12g agar, 0.5~1g CaCl2, 30~33g sucrose, 60~65g glucose, 1~3g potassium sorbate and
5% butyl p-hydroxybenzoate solution of 15ml；More preferably include 77.7g maize flour, 32.19g yeast, 10.6g agar,
0.726g CaCl2, 31.62g sucrose, 5% butyl p-hydroxybenzoate solution of 63.2g glucose, 2g potassium sorbate and 15ml.
In the present invention, in the solid medium containing preferably comprising the solvents of dissolved contaminants, the type of the solvent, content and contain
There are type, the content of the solvent in the solid medium of pollutant identical.
In the present invention, the Drosophila melanogaster is placed in the condition cultivated on solid medium and Drosophila melanogaster is placed in containing dirt
The condition cultivated on the solid medium of dye object is identical, and details are not described herein.
In the present invention, the significant difference P value is less than 0.01.
Technical solution provided by the invention is described in detail below with reference to embodiment, but they cannot be understood
For limiting the scope of the present invention.
Embodiment 1
Preparation of reagents: triclosan, purity 97% are purchased from Shanghai Mike woods biochemical technology Co., Ltd, use dimethyl sulfoxide
Dissolution.Isometric various concentration stock solution is added in standard medium to prescribed concentration, is added isometric two in blank control group
Methyl sulfoxide.
Exposure condition: 5 experimental groups of setting, every group 5 parallel；Blank control, triclosan (10,100,250,500mg/
L).It is respectively put into 30 female male drosophilas newly sprouted wings in each parallel laboratory test group and carries out individually exposure culture, is placed in standard culture
Under the conditions of.
Pattern detection: measuring female male drosophila when contamination culture 1d, 3d, 5d, 7d, 9d, 11d, 13 and 15d and creep ability, will
Each group drosophila is placed in 100mL graduated cylinder with carbon dioxide narcosis after contamination, carries out climbing survey after each group drosophila is completely awake
Examination measures the drosophila quantity creeped in each group drosophila 10s more than 10cm, calculated line rate.The line of crossing of female fly the results are shown in Table 1 and figure
1, the line of crossing of male fly the results are shown in Table 2 and Fig. 2.
The female fly of table 1 gets over line rate result
The male fly of table 2 gets over line rate result
It is indicated above that triclosan influences the climbing ability of drosophila, Initial stage of culture triclosan has certain stimulation to drosophila,
Climbing ability compared with control group compared to being risen, but under the conditions of the extension high concentration triclosan of incubation time male and female adult flies climb
Ability is climbed compared with control group compared to being remarkably decreased, balance exercise reduced capability.
Embodiment 2
Preparation of reagents: diisononyl phthalate (DINP), purity 99.5% are purchased from Aladdin chemical reagents corporation,
It is dissolved with dehydrated alcohol.Isometric various concentration stock solution is added in standard medium to prescribed concentration, in blank control group plus
Enter isometric dehydrated alcohol.
Exposure condition: 5 experimental groups of setting, every group 5 parallel；Blank control, DINP (0.1%, 0.2%, 0.5%,
1.0%).It is put into the female male drosophila that 30 are newly sprouted wings in each parallel laboratory test group to carry out exposing culture, is placed in Standard culture conditions
Under.
Pattern detection: female male drosophila is measured when contamination culture 1d, 5d, 10d and 15d and is creeped ability, by each group fruit after contamination
Fly is placed in 100mL graduated cylinder with carbon dioxide narcosis, is carried out climbing test after each group drosophila is completely awake, is measured each group fruit
The drosophila quantity creeped in fly 10s more than 10cm, calculated line rate.The line of crossing of female fly the results are shown in Table 3 and Fig. 3, and male fly crosses line
It the results are shown in Table 4 and Fig. 4.
The female fly of table 3 gets over line rate result
The male fly of table 4 gets over line rate result
Thus, it is indicated, that DINP influences the climbing ability of drosophila, male and female adult flies climbing ability control under high concentration DINP
Group is compared to being remarkably decreased, and low concentration DINP has certain stimulation, and climbing ability is risen but ability reduces over time.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (8)
1. a kind of method of the toxicity using Drosophila melanogaster behavior stress reaction evaluation pollutant, which is characterized in that including following
Step:
1) Drosophila melanogaster is placed in 1~15d of culture on the solid medium containing pollutant, detection Drosophila melanogaster is climbed in 10s
Row is more than the quantity of 10cm, and line rate is obtained；
2) Drosophila melanogaster is placed in 1~15d of culture on solid medium, detection Drosophila melanogaster is creeped in 10s more than 10cm's
Line rate is obtained in quantity；
3) when significant difference occurs in the line rate excessively that line rate and step 2) excessively that the step 1) obtains obtain, show that pollutant has
It is toxic.
2. the method according to claim 1, wherein the pollutant includes triclosan and/or phthalic acid
Dinonyl.
3. method according to claim 1 or 2, which is characterized in that described to contain three when the pollutant is triclosan
The concentration of triclosan is 10~500mg/L in the raw solid medium of chlorine.
4. method according to claim 1 or 2, which is characterized in that when the pollutant is diisononyl phthalate
When, the volumetric concentration of diisononyl phthalate is 0.1 in the solid medium containing diisononyl phthalate
~1%.
5. the method according to claim 1, wherein the solid medium containing pollutant is molten with water
Agent, every liter further include: 70~80g maize flour, 30~35g yeast, 8~12g agar, 0.5~1g CaCl2, 30~33g sucrose,
5% butyl p-hydroxybenzoate solution of 60~65g glucose, 1~3g potassium sorbate and 15ml.
6. every liter includes: 70 the method according to claim 1, wherein the solid medium takes water as a solvent
~80g maize flour, 30~35g yeast, 8~12g agar, 0.5~1g CaCl2, 30~33g sucrose, 60~65g glucose, 1
5% butyl p-hydroxybenzoate solution of~3g potassium sorbate and 15ml.
7. the method according to claim 1, wherein the female fly of the Drosophila melanogaster and male fly are individually cultivated.
8. the method according to claim 1, wherein the temperature that the feeding condition includes: the culture is 24
~26 DEG C, the Light To Dark Ratio of the culture is 12h:12h；The ambient humidity of the culture is 50~60%.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201910533336.7A CN110235862A (en) | 2019-06-19 | 2019-06-19 | A method of utilizing the toxicity of Drosophila melanogaster behavior stress reaction evaluation pollutant |
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| CN201910533336.7A CN110235862A (en) | 2019-06-19 | 2019-06-19 | A method of utilizing the toxicity of Drosophila melanogaster behavior stress reaction evaluation pollutant |
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