CN104823924B - Toxicity test method of the systemic insecticide to green plant bug - Google Patents
Toxicity test method of the systemic insecticide to green plant bug Download PDFInfo
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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
-
- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06F—ELECTRIC DIGITAL DATA PROCESSING
- G06F17/00—Digital computing or data processing equipment or methods, specially adapted for specific functions
- G06F17/10—Complex mathematical operations
- G06F17/18—Complex mathematical operations for evaluating statistical data, e.g. average values, frequency distributions, probability functions, regression analysis
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- Life Sciences & Earth Sciences (AREA)
- Data Mining & Analysis (AREA)
- General Physics & Mathematics (AREA)
- Environmental Sciences (AREA)
- Theoretical Computer Science (AREA)
- Computational Mathematics (AREA)
- Mathematical Physics (AREA)
- Mathematical Analysis (AREA)
- Mathematical Optimization (AREA)
- Pure & Applied Mathematics (AREA)
- Probability & Statistics with Applications (AREA)
- Bioinformatics & Computational Biology (AREA)
- Algebra (AREA)
- Evolutionary Biology (AREA)
- Databases & Information Systems (AREA)
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- General Engineering & Computer Science (AREA)
- Operations Research (AREA)
- Animal Behavior & Ethology (AREA)
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Abstract
The present invention relates to a kind of toxicity test method of effective evaluation systemic insecticide to green plant bug, comprise the following steps:Making, feeding, test statistics that prepared by man-made feeds, subjects prepare, feed is mixed, mix medicine feed bag etc., toxicity test method of the present invention overcomes defect of the prior art, medicine feeding is mixed using the good man-made feeds of stable, uniformity, and simulate green plant bug take food state naturally, it is finally reached the purpose of the objective correct killing effect for evaluating systemic insecticide, be conducive to filtering out and efficient insecticide is killed to green plant bug, to instruct the preventing and treating of crop field green plant bug to provide technical support.
Description
Technical field
The invention belongs to agricultural chemical insecticide field, and in particular to a kind of poison of effective evaluation systemic insecticide to green plant bug
Power assay method.
Background technology
Green plant bugApolygus lucorum(Meyer-D ü r) belongs to Semiptera (Hemiptera) Miridae (Miridae)
Apolygus (Apolygus), its host range is extensive, it has been reported that more than 50 sections kind more than 200, including cotton, jujube, grape, cherry
A variety of important crops such as peach, apple, tea tree.In recent years, due to the adjustment of agricultural structure, fruit tree and vegetable cultivation area
Increase considerably, abundant host plant and suitable hibernacle are provided for green plant bug, simultaneously because Bt transgenic cotton
A large amount of plantations, the usage amount for preventing and treating the insecticide of bollworm is reduced, and causes sucking number of pest to steeply rise, and it is long-term with
Turn into the primary pest (Wu in cotton field to be constantly in the green plant bug of back burneret al., 2008; Lu et al.,
2010b)。
At present, it is still the Main Means for controlling green plant bug population quantity to spray chemical insecticide, and domestic scholars are in insecticide
Screening and alternative aspect expand many work, but most of application bottle embrane methods and drop method have filtered out part effectively preventing and treating
Contact efficacy can be evaluated in the medicament of fleahopper, this two methods.For example, according to bottle embrane method and drop method measure to green plant bug
Virulence effect, organophosphor and chrysanthemum ester insecticide are above anabasine insecticide（Zhanget al, 2009；Lee state equality,
2009；Men Xingyuan etc., 2010；Li Yaofa etc., 2007）.Therefore, in actual applications, chlopyrifos, horse traction sulphur are typically all recommended
The stronger medicament of the actions of contace poison such as phosphorus, Biphenthrin.But inhaled because green plant bug adult and nymph have to hide to pierce in cotton bract
The habit caused harm, causes contacting pests medicament heterogeneity, adds the difficulty to its effective improvement.Anabasine insecticide is acted on
Based on being inhaled within mode, if systemic insecticide is good to green plant bug prevention effect, endangered even if fleahopper is hidden in cotton bract
Evil, can also be killed after taking food.It would therefore be highly desirable to propose a kind of toxicity test method of systemic insecticide to green plant bug, come
The objective correct effect for evaluating systemic insecticide, to instruct the preventing and treating of crop field green plant bug to provide technical support.
The content of the invention
In view of the above-mentioned problems, the present invention provides a kind of systemic insecticide to the toxicity test method of green plant bug, can be objective
The correct killing effect for evaluating systemic insecticide, available for the screening and compounding for killing fleahopper medicament, to instruct crop field green blind
The preventing and treating of stinkbug provides technical support.
In order to solve the above technical problems, the present invention is achieved through the following technical solutions：
A kind of toxicity test method of systemic insecticide to green plant bug is designed, is comprised the following steps:
（1）It is prepared by man-made feeds
1. in parts by weight, first take 20~55 parts of sucrose, 15~55 parts of 50% hydromel, 1~11 part of 10% acetic acid with 70~
Heat and boil after 90 parts of water mixing, 1~11 part of beer yeast powder is added in heating process, then pour into the egg 200~355 of mixing
Part, untill ceaselessly stirring is when the golden yellow into uniformity；2. 200~450 parts of yolk, analysis for soybean powder 10~45 are taken again
Part, 50~150 parts of wheat, 850~950 parts of 100~250 parts of butter bean coarse powder and water is uniform mix after, 121 °C, high pressure goes out
15~25min of bacterium；3. 10~45 parts of soybean lecithin, 10~25 parts of B B-complex, streptomycin sulphate 0.06~0. 30 are taken again
Part, 0.03~0.30 part of cephalosporin analog antibiotic, 0.7~1.5 part of 37% formalin, 0.7~1.5 part of propionic acid, benzoic acid 3.0
~6.5 parts with sterilized water 650~670 parts uniformly mix；4. the compound after finally 2. step is sterilized, which takes out, is cooled to 50 °C
When, be separately added into step 1. with step 3. gained compound, after stirring；
The nutrition of above-mentioned man-made feeds, which is constituted, is adapted to growing for green plant bug, and the stability, repeatable that feed is raised
Property it is strong, be conducive to ensureing the uniformity that green plant bug takes food, to obtain true and reliable virulence determination data；
（2）Subjects prepare：
Green plant bug nymph was raised with kidney bean to the emerging adult stage, any medicament is not during which contacted, it is standby as test worm
With；
（3）Feed is mixed：
Certain density systemic insecticide mother liquor is prepared by solvent of acetone, then systemic insecticide mother liquor is diluted to
The ratio such as series or equal difference mass concentration gradient, take 0.5~1mL of dilution of each concentration of gained to be uniformly blended into 50~100g respectively
In the man-made feeds, 4 DEG C of Cord bloods are sealed, it is standby as test group feed；Added in the man-made feeds of phase homogenous quantities
0.5~1mL acetone, is stirred, and seals 4 DEG C of Cord bloods, standby as blank control group feed；
（4）The making of mixed medicine feed bag：
5cm × 5cm bags are made with Parafilm films, test group and control group feed are poured into respectively, correspondence is made in sealing
Mixed medicine feed bag, it is standby；
（5）Feeding：
By each mixed medicine feed bag Uniform Tension to original 2 times（Pierce and inhale beneficial to green plant bug lancet）, it is put into corresponding 150~
In 300mL cups, then the green plant bug adult Nature enemy 5h in emergence 10d is chosen, random being linked into 10 every glass is described
In cup, rim of a cup is covered with gauze, is placed on the bar that temperature is 25~27 DEG C, relative humidity is 75~85%, illumination L/D=16h/8h
Under part, viewing test group and control group test worm death condition after 48h；
（6）Test statistics：
Medicament toxicity regression curve is fitted, LC is searched or calculate50Value（Median lethal dose）And 95% confidence limit.It can use
Polo statistical softwares carry out statistics calculating.
For above-mentioned toxicity test method, the ratio such as step (3) described series or equal difference mass concentration gradient are 5~7
Deng ratio or equal difference mass concentration gradient.
For above-mentioned toxicity test method, the systemic insecticide content in the test group feed is controlled in kill rate
In the range of 10~90%.
For the control of the systemic insecticide content, pilot study can be first done：Systemic insecticide configuration is certain
The acetone soln of concentration, according to step（3）Add man-made feeds to mix, then choose at the green plant bug adult starvation in emergence 10d
5h is managed as test worm, is accessed in man-made feeds after test worm 48h, according to death condition, up-regulation or downward insecticide concentration make to kill
The ratio of dead test worm accounts for 10~90%.
The positive beneficial effect of the present invention：
Toxicity test method of the present invention overcomes defect of the prior art, using the good man-made feeds of stable, uniformity
Mixed medicine feeding, and simulate the killing for taking food state naturally, being finally reached objective correct evaluation systemic insecticide of green plant bug
The purpose of effect, is conducive to filtering out and kills efficient insecticide to green plant bug, to instruct the preventing and treating of crop field green plant bug to provide skill
Art is supported.
Embodiment
Technical solution of the present invention is further illustrated below in conjunction with specific embodiment.It is raw materials used in following embodiment, such as
Then it is commercially available without special instruction.
Embodiment 1
A kind of systemic insecticide comprises the following steps to the toxicity test method of green plant bug:
（1）It is prepared by man-made feeds:
Raw material：Mix egg（Egg white, yolk mixing）300 parts, 40 parts of sucrose, 6 parts of brewer's yeast, 50% hydromel 30
Part, 350 parts of yolk, 100 parts of wheat, 200 parts of butter bean coarse powder, 30 parts of soy meal, 30 parts of soybean lecithin, B B-complex
20 parts, 1640 parts of water, 0.15 part of streptomycin sulphate, 0.15 part of cephalosporin analog antibiotic, 6 parts of 10% acetic acid, 37% formalin 1.2
Part, 1.2 parts of propionic acid, 5 parts of benzoic acid, the preparation method of the man-made feeds comprises the following steps：1）First by sucrose, hydromel,
10% acetic acid is heated after being mixed with 80 parts of water to be boiled, and beer yeast powder is added in heating process, then pours into the egg of mixing, is not stopped
Stirring, untill when compound is into golden yellow and uniformity；2）By yolk, analysis for soybean powder, wheat, butter bean coarse powder and water
After 900 parts of uniform mixing, 121 °C, autoclaving 20min；3）Again by soybean lecithin, B B-complex, antibiotic, remaining prevents
Rotten agent is uniformly mixed for 660 parts with sterilized water；4）By step 2）When compound taking-up after sterilizing is cooled to 50 °C, respectively
Add step 1）With step 3）Gained compound, after stirring；
（2）Subjects prepare:
Green plant bug nymph was raised with kidney bean to the emerging adult stage, any medicament is not during which contacted, it is standby as test worm
With；
（3）Feed is mixed：
Certain density insecticide mother liquor is prepared by solvent of acetone, then insecticide mother liquor is diluted to the ratio such as series or waited
Difference quality measurement concentration gradient（It is shown in Table 1）, take the dilution 1mL of each concentration of gained to be uniformly blended into man-made feeds described in 100g respectively
In, 4 DEG C of Cord bloods are sealed, it is standby as test group feed；1mL acetone is added in the man-made feeds of phase homogenous quantities, fully
Stir, seal 4 DEG C of Cord bloods, it is standby as blank control group feed；
The insecticide species of table 1 and mass concentration gradient are set
（4）The making of mixed medicine feed bag：
5cm × 5cm bags are made with Parafilm films, test group and control group feed are poured into respectively, correspondence is made in sealing
Mixed medicine feed bag, it is standby；
（5）Feeding：
By each mixed medicine feed bag Uniform Tension to original 2 times, it is put into corresponding 250mL cups, then choose emergence 10d
Interior green plant bug adult Nature enemy 5h, random is linked into the cup with 10 every glass, and rim of a cup is covered with gauze, is placed on
Under conditions of temperature is 25~27 DEG C, relative humidity is 75~85%, illumination L/D=16h/8h, viewing test group and control after 48h
Group test worm death condition (with tweezers or other hard thing disturbance adults, death is judged as if it can not normally creep), each processing
If 3 repetitions；
（6）Test statistics：
Medicament toxicity regression curve is fitted, LC is searched or calculate50Value and 95% confidence limit, are all provided with control in each experiment
Reason, is analyzed using polo-pc (Leora, 1987) software；
（7）Statistical result：
As can be seen from Table 2, organophosphorus insecticides be malathion to the virulence of green plant bug adult order, chlopyrifos,
Phoxim, orthene, Rogor, LC50Value is respectively 0.108,0.173,0.745,0.758,4.425 μ g/mL, and chrysanthemum esters are killed
Worm agent is to green plant bug virulence LC50Value respectively Biphenthrin, decis, Cyhalothrin, effective cypermethrin, 0.062,
1.100、3.163、6.479μg/mL；Virulence Acetamiprid, Nitenpyram, imidacloprid, the Diacloden of anabasine insecticide, LC50
Value is respectively 0.034,0.025,0.011,0.051 μ g/mL.According to LC50Be worth confidential interval it is overlapping whether, anabasine insecticide
LC50Value is significantly lower than organophosphorus insecticide and chrysanthemum ester insecticide, and multiple differs about 10~100 times, illustrates that anabasine is killed
The absorbability effect of worm agent is fabulous.
The test statistics result of 2 embodiment of table 1
Comparative example 1（Bottle embrane method）
A kind of systemic insecticide comprises the following steps to the toxicity test method of green plant bug:
（1）It is prepared by man-made feeds:
Be the same as Example 1.
（2）Subjects prepare:
Green plant bug nymph was raised with kidney bean to the emerging adult stage, any medicament is not during which contacted, it is standby as test worm
With；
（3）Use acetone as organic solvent and prepare certain density mother liquor, mother liquor is diluted to serial matter according still further to table 1
Measure concentration gradient；The decoction 0.5mL of mass concentration to be measured is moved into 20mL scintillation vials with liquid-transfering gun, blank control is without decoction
0.5mL acetone, by scintillation vial lie on Sandwich roaster rotate, organic solvent-acetone is volatilized at room temperature, formed in bottle inwall
The medicine film of uniformity.Green plant bug adult within two is sprouted wings 10 days is put into the scintillation vial with medicine film, and is put into thick about
3mm fresh kidney bean 1 is as food, and each concentration surveys 10 adults, and bottleneck is clogged with tampon, and it is 25~27 to be placed on temperature
DEG C, relative humidity be 75~85%, under conditions of illumination L/D=16h/8h, the death rate is determined after 24h and (uses tweezers or other hard things
Adult is disturbed, death is judged as if it can not normally creep), each processing sets 3 repetitions；
（4）Test statistics：
Medicament toxicity regression curve is fitted, LC is searched or calculate50Value and 95% confidence limit；
（5）Statistical result：
Using bottle embrane method, as can be seen from Table 3, organophosphorus insecticides is chlopyrifos to the virulence order of green plant bug adult
>Malathion, phoxim>Acetyl first pacifies phosphorus>Rogor, LC50Value is respectively 1.803,3.486,4.659,20.06,27.602 μ g/
ML, chrysanthemum ester insecticide is Biphenthrin to the virulence order of green plant bug adult>Cyhalothrin>Decis>Efficient chlorine
Cyano chrysanthemate, LC50Value is respectively 0.698,48.453,63.333,81.703 μ g/mL；The virulence alkene pyridine worm of anabasine insecticide
Amine, imidacloprid, Diacloden, Acetamiprid, LC50Value is respectively 428.205,472.644,370.282,425.657 μ g/mL.According to
LC50Be worth confidential interval it is overlapping whether, the LC of anabasine insecticide50Value is significantly higher than organophosphorus insecticide and chrysanthemum esters desinsection
Agent, illustrates that the contact efficacy of anabasine insecticide is poor.
The test statistics result of 3 comparative example of table 1
To sum up, embodiment 1 and comparative example 1 are contrasted, it can be seen from bottle embrane method toxicity test result and organophosphor
Class, chrysanthemum ester insecticide are compared, and anabasine insecticide is poor to the Contact toxicity of green plant bug.By the toxicity test knot of embodiment 1
Fruit can be seen that compared with organic phosphates, chrysanthemum ester insecticide, systemic action of the anabasine insecticide to green plant bug adult
It is good.
Embodiment 2
A kind of systemic insecticide comprises the following steps to the toxicity test method of green plant bug:
（1）It is prepared by man-made feeds:
Raw material：Mix 200 parts of egg, 20 parts of sucrose, 1 part of brewer's yeast, 15 parts of 50% hydromel, 200 parts of yolk, wheat
50 parts of embryo, 100 parts of butter bean coarse powder, 10 parts of soy meal, 10 parts of soybean lecithin, 10 parts of B B-complex, 1570 parts of water, sulphur
Sour 0.06 part of streptomysin, 0.03 part of cephalosporin analog antibiotic, 1 part of 10% acetic acid, 0.7 part of 37% formalin, 0.7 part of propionic acid, benzene first
3.0 parts of acid, the preparation method of the man-made feeds comprises the following steps：1）First by sucrose, hydromel, 10% acetic acid and 70 parts of water
Heating is boiled after mixing, and beer yeast powder is added in heating process, then pours into the egg of mixing, is ceaselessly stirred, to compound
Untill during golden yellow and uniformity；2）Yolk, analysis for soybean powder, wheat, butter bean coarse powder are uniformly mixed for 850 parts with water
Afterwards, 121 °C, autoclaving 15min；3）Again by soybean lecithin, B B-complex, antibiotic, remaining preservative and sterilized water
650 parts of uniform mixing；4）By step 2）When compound taking-up after sterilizing is cooled to 50 °C, step 1 is separately added into）With
Step 3）Gained compound, after stirring；
（2）Subjects prepare:
Green plant bug nymph was raised with kidney bean to the emerging adult stage, any medicament is not during which contacted, it is standby as test worm
With；
（3）Feed is mixed：
Certain density systemic insecticide mother liquor is prepared by solvent of acetone, then systemic insecticide mother liquor is diluted to
The ratio such as series or equal difference mass concentration gradient, take the dilution 0.5mL of each concentration of gained to be uniformly blended into described in 50g manually respectively
In feed, 4 DEG C of Cord bloods are sealed, it is standby as test group feed；0.5mL third is added in the man-made feeds of phase homogenous quantities
Ketone, is stirred, and seals 4 DEG C of Cord bloods, standby as blank control group feed；
（4）The making of mixed medicine feed bag：
5cm × 5cm bags are made with Parafilm films, test group and control group feed are poured into respectively, correspondence is made in sealing
Mixed medicine feed bag, it is standby；
（5）Feeding：
By each mixed medicine feed bag Uniform Tension to original 2 times, it is put into corresponding 300mL cups, then choose emergence 10d
Interior green plant bug adult Nature enemy 5h, random is linked into the cup with 10 every glass, and rim of a cup is covered with gauze, is placed on
Under conditions of temperature is 25~27 DEG C, relative humidity is 75~85%, illumination L/D=16h/8h, viewing test group and control after 48h
Group test worm death condition (with tweezers or other hard thing disturbance adults, death is judged as if it can not normally creep), each processing
If 3 repetitions；
（6）Test statistics：
Medicament toxicity regression curve is fitted, LC is searched or calculate50Value and 95% confidence limit, are all provided with control in each experiment
Reason, is analyzed using polo-pc (Leora, 1987) software.
Embodiment 3
A kind of systemic insecticide comprises the following steps to the toxicity test method of green plant bug:
（1）It is prepared by man-made feeds:
Raw material：Mix 355 parts of egg, it is 55 parts of sucrose, 11 parts of brewer's yeast, 55 parts of 50% hydromel, 450 parts of yolk, small
150 parts of wheat germ, 250 parts of butter bean coarse powder, 45 parts of soy meal, 45 parts of soybean lecithin, 25 parts of B B-complex, 1710 parts of water,
0. 30 parts of streptomycin sulphate, 0.30 part of cephalosporin analog antibiotic, 11 parts of 10% acetic acid, 1.5 parts of 37% formalin, 1.5 parts of propionic acid,
6.5 parts of benzoic acid, the preparation method of the man-made feeds comprises the following steps：1）First by sucrose, hydromel, 10% acetic acid and 90
Heat and boil after part water mixing, beer yeast powder is added in heating process, then pour into the egg of mixing, ceaselessly stir, it is extremely mixed
Untill when golden yellow and uniformity is expected into conjunction；2）950 parts of yolk, analysis for soybean powder, wheat, butter bean coarse powder and water is uniform mixed
After conjunction, 121 °C, autoclaving 25min；3）Again by soybean lecithin, B B-complex, antibiotic, remaining preservative with it is sterile
The uniform mixing of 670 parts of water；4）By step 2）When compound taking-up after sterilizing is cooled to 50 °C, step 1 is separately added into）
With step 3）Gained compound, after stirring；
（2）Subjects prepare:
Green plant bug nymph was raised with kidney bean to the emerging adult stage, any medicament is not during which contacted, it is standby as test worm
With；
（3）Feed is mixed：
Certain density systemic insecticide mother liquor is prepared by solvent of acetone, then systemic insecticide mother liquor is diluted to
The ratio such as series or equal difference mass concentration gradient, take the dilution 0.75mL of each concentration of gained to be uniformly blended into people described in 75g respectively
In work feed, 4 DEG C of Cord bloods are sealed, it is standby as test group feed；0.75mL is added in the man-made feeds of phase homogenous quantities
Acetone, is stirred, and seals 4 DEG C of Cord bloods, standby as blank control group feed；
（4）The making of mixed medicine feed bag：
5cm × 5cm bags are made with Parafilm films, test group and control group feed are poured into respectively, correspondence is made in sealing
Mixed medicine feed bag, it is standby；
（5）Feeding：
By each mixed medicine feed bag Uniform Tension to original 2 times, it is put into corresponding 150mL cups, then choose emergence 10d
Interior green plant bug adult Nature enemy 5h, random is linked into the cup with 10 every glass, and rim of a cup is covered with gauze, is placed on
Under conditions of temperature is 25~27 DEG C, relative humidity is 75~85%, illumination L/D=16h/8h, viewing test group and control after 48h
Group test worm death condition (with tweezers or other hard thing disturbance adults, death is judged as if it can not normally creep), each processing
If 3 repetitions；
（6）Test statistics：
Medicament toxicity regression curve is fitted, LC is searched or calculate50Value and 95% confidence limit, are all provided with control in each experiment
Reason, is analyzed using polo-pc (Leora, 1987) software.
Claims (2)
1. a kind of systemic insecticide is to the toxicity test method of green plant bug, it is characterised in that comprise the following steps:
（1）It is prepared by man-made feeds：
1. in parts by weight, 20~55 parts of sucrose, 15~55 parts of 50% hydromel, 1~11 part and 70~90 parts of 10% acetic acid are first taken
Heat and boils after water mixing, 1~11 part of beer yeast powder of addition in heating process, then pour into 200~355 parts of the egg of mixing,
Untill ceaselessly stirring is when the golden yellow into uniformity；2. take 200~450 parts of yolk again, it is 10~45 parts of analysis for soybean powder, small
850~950 parts of 50~150 parts of wheat germ, 100~250 parts of butter bean coarse powder and water is uniform mix after, 121 °C, autoclaving 15~
25min；3. 10~45 parts of soybean lecithin, 10~25 parts of B B-complex, streptomycin sulphate 0.06~0. 30 part, head are taken again
0.03~0.30 part of spore class antibiotic, 0.7~1.5 part of 37% formalin, 0.7~1.5 part of propionic acid, benzoic acid 3.0~6.5
650~670 parts with sterilized water of part uniformly mixes；4. when the compound taking-up after finally 2. step is sterilized is cooled to 50 °C, point
Not Jia Ru step 1. with step 3. gained compound, after stirring；
（2）Subjects prepare：
Green plant bug nymph was raised with kidney bean to the emerging adult stage, any medicament is not during which contacted, it is standby as test worm；
（3）Feed is mixed：
Certain density systemic insecticide mother liquor is prepared by solvent of acetone, then systemic insecticide mother liquor is diluted to series
Deng ratio or equal difference mass concentration gradient, 0.5~1.0mL of dilution of each concentration of gained is taken to be uniformly blended into 50~100g institutes respectively
State in man-made feeds, seal 4 DEG C of Cord bloods, it is standby as test group feed；0.5 is added in the man-made feeds of phase homogenous quantities
~1mL acetone, is stirred, and seals 4 DEG C of Cord bloods, standby as blank control group feed；
Systemic insecticide content in the test group feed is controlled in the range of kill rate 10~90%；
The control method of the systemic insecticide content is：Systemic insecticide is configured into finite concentration acetone soln, according to
Step（3）Add man-made feeds to mix, then choose green plant bug adult Nature enemy 5h in emergence 10d as test worm, artificial
Being accessed in feed after test worm 48h, according to death condition, up-regulation or lower insecticide concentration, the ratio for making kill test worm accounts for 10~
90%；
（4）The making of mixed medicine feed bag：
5cm × 5cm bags are made with Parafilm films, test group and control group feed are poured into respectively, sealing is made corresponding mixed
Medicine feed bag, it is standby；
（5）Feeding：
By each mixed medicine feed bag Uniform Tension to original 2 times, it is put into corresponding 150~300mL cups, then choose emergence
Green plant bug adult Nature enemy 5h in 10d, random is linked into the cup with 10 every glass, and rim of a cup is covered with gauze,
It is 25~27 DEG C, under conditions of relative humidity is 75~85%, illumination L/D=16h/8h to be placed on temperature, after 48h viewing test group and
Control group test worm death condition；
（6）Test statistics：
Medicament toxicity regression curve is fitted, LC is searched or calculate50Value and 95% confidence limit.
2. toxicity test method according to claim 1, it is characterised in that：Ratio or the equal difference matter such as step (3) described series
It is the ratios such as 5~7 or equal difference mass concentration gradient to measure concentration gradient.
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