CN105850898A - Method for measuring acute and chronic toxicity of drug against varroa jacobsoni - Google Patents
Method for measuring acute and chronic toxicity of drug against varroa jacobsoni Download PDFInfo
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- CN105850898A CN105850898A CN201610375448.0A CN201610375448A CN105850898A CN 105850898 A CN105850898 A CN 105850898A CN 201610375448 A CN201610375448 A CN 201610375448A CN 105850898 A CN105850898 A CN 105850898A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
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Abstract
The invention discloses a method for measuring acute and chronic toxicity of a drug against varroa jacobsoni. The method comprises the following steps: gently spraying the to-be-measured liquid onto the surfaces of healthy bee larvae; transferring the bee larvae from the inside of a larva culture plate into a bee pupa culture cup with the liquid-infiltrated filter paper piece at the bottom; collecting the to-be-measured varroa jacobsoni; introducing a varroa jacobsoni into the bee pupa culture cups of each treatment group and control group, and sealing the cup rims of the bee pupa culture cups with ventilated transparent films to prevent the introduced varroa jacobsoni from escaping the bee pupa culture cups; putting the bee pupa culture cups into an incubator for culturing, wherein the temperature is 34.5 DEG C, and the relative humidity is 75%; counting the deaths of varroa jacobsoni 48h or 72h later, calculating the death rate and analyzing the acute toxicity of the drug against varroa jacobsoni; or continuously recording the survival conditions of the varroa jacobsoni every day, counting the survival time of each varroa jacobsoni, and analyzing the chronic toxicity of the drug against varroa jacobsoni. The method disclosed by the invention has the advantages of controllable experiment conditions, high survival rate of the varroa jacobsoni, long survival time and accurate and credible result.
Description
Technical field
The present invention relates to measure the medicine method to insect toxicity, particularly relate to a kind of medicine that measures to big honeybee
The method of the acute and chronic toxicity of demodicid mite.
Background technology
Apis is important economic insects, has the biggest economic worth and the ecological value, and Apis was both provided that
Useful bee product can be pollinated for plant again, and varoa mite is the most serious Apis harmful animal of harm apiculture.
Varoa mite is the most serious Apis harmful animal of harm world's apiculture, due to life-time service pyrethroid
Class acaricide, varoa mite creates serious Drug resistance, and the acaricidal screening of new and effective green is extremely urgent.
And lack the acaricide standard method to varoa mite toxicity test in the world.At present, two kinds of sides of general employing
Method, one is bee colony toxicity test method, and another kind is Toxicity Determination method.Both approaches all has necessarily
Shortcoming, it is impossible to the Accurate Determining acaricide virulence to varoa mite.Bee colony dispenser method: (1) uncontrollable examination
Test condition, affected by factors such as environment and bee colony group's gesture relatively big, cause systematic error big；(2) reagent dosage
It not the dosage of the actual contact of honeybee demodicid mite, cause result inaccurate；(3) the honeybee demodicid mite parasitic rate between test bee colony
Inconsistent, differ greatly in causing process group；(4) bee colony test is wasted time and energy.Toxicity Determination method is
Honeybee demodicid mite access inwall is evenly coated with in culture dish or the vial of medicine to be measured, measures honeybee demodicid mite mortality rate；Or
Honeybee demodicid mite is put into culture dish, then sprays medicine to be measured facing to honeybee demodicid mite, measure honeybee demodicid mite mortality rate.Indoor virulence
The shortcoming of algoscopy is, lacks the factors such as food due to honeybee demodicid mite, causes matched group honeybee demodicid mite mortality rate high, result
Insincere, also cannot measure the medicine chronic toxicity to varoa mite.
Summary of the invention
The technical problem to be solved in the present invention is to provide that experiment condition is controlled, honeybee demodicid mite survival rate high, the time-to-live
Long, the method for result accurate believable mensuration medicine toxicity acute and chronic to varoa mite.
The method of varoa mite toxicity is comprised the steps: by the medicine that measures of the present invention
1) medicine to be measured is arranged according to process group, be configured to different concentration liquid to be measured with solvent, will treat
The healthy bee larvae body surface that measured quantity medicinal liquid is sprayed on gently, is then placed in one 34.5 DEG C by bee larvae
Pupa Apis after preheating is cultivated in cup；Matched group solvent processes；Lay the denseest bottom described Pupa Apis cultivation cup
The filter paper dried after degree medicine infiltration；
2) collecting varoa mite to be measured from the foster demodicid mite spleen prepared, the Pupa Apis of each process group and matched group is cultivated
One varoa mite of access in cup, and the rim of a cup of Pupa Apis cultivation cup is sealed with ventilative transparent membrane, Pupa Apis is trained
Foster cup is put in the hole of Pupa Apis culture plate, one, every hole, puts in incubator by Pupa Apis culture plate, and temperature is
34.5 DEG C, relative humidity is 75%；
3) add up the different disposal group varoa mite death toll of different time sections, calculate mortality rate, analyze medicine
Acute toxicity to varoa mite；Or every day records the survival condition of test honeybee demodicid mite continuously, add up every honeybee demodicid mite
Time-to-live, analyze the medicine chronic toxicity to varoa mite；Wherein, the honeybee demodicid mite number/access of mortality rate=death
Honeybee demodicid mite number.
In described method, the bee larvae of described health is in capping larval phase.
In described method, the bee larvae of described health is the larva of Apis mellifera (Apis melifera).
In described method, the solvent of medicine to be measured can be acetone.
In described method, medicine to be measured can be coumafos (Coumaphos).
In described method, the cultural method of the bee larvae of described health is as follows: with laying eggs, controller controls honeybee
Wang Yi opens and lays eggs on sky Nidus Vespae, discharges queen bee after 24h, after 3 days with Moving needle for bee-keeping larva is transferred to equipped with
In the larva culture plate being provided with some holes accommodating larva of 34.5 DEG C of preheatings of feedstuff, every hole moves into one
Bee larvae；Larva culture plate is placed in incubator, and temperature is 34.5 DEG C, relative humidity 95%, even
Continue and feed 6 days；Within 1st day, feed 10 μ L feedstuff A, within the 2nd day, feed 10 μ L feedstuff A, within the 3rd day, raise
Feed 20 μ L feedstuff B, within the 4th day, feed 30 μ L feedstuff C, within the 5th day, feed 40 μ L feedstuff C, the 6th day
Feed 50 μ L feedstuff C, obtain covering larval phase health bee larvae；Described feedstuff A is by containing weight hundred
Glucose, weight percentage that Lac regis apis that point content is 44.25%, weight percentage are 5.3% are
The fructose of 5.3%, weight percentage be 0.90% yeast extract and weight percentage be 44.25%
Water composition；Described feedstuff B is contained by containing the Lac regis apis that weight percentage is 42.95%, weight percent
Fructose, weight percentage that glucose that amount is 6.40%, weight percentage are 6.40% are 1.30%
Yeast extract and water that weight percentage is 42.95% composition；Described feedstuff C is by containing weight hundred
Glucose, the weight percentage that Lac regis apis that point content is 50.00%, weight percentage are 9.00%
Be 9.00% fructose, weight percentage be 2.00% yeast extract and weight percentage be 30.00%
Water composition.Concrete, the preparation method of the Pupa Apis cultivation cup after 34.5 DEG C of preheatings is as follows: medicine to be measured is used
Solvent is configured to different concentration liquid to be measured, is dripped to by quantitative liquid medicine on the filter paper got ready, and filter paper is whole
Soak, then the filter paper of coating is placed in super-clean bench after solvent volatilization is dried, put into Pupa Apis training
Supporting the bottom of cup, each Pupa Apis cultivates one filter paper of cup, Pupa Apis is cultivated cup and is positioned over Pupa Apis culture plate
In hole, Pupa Apis culture plate is placed in incubator, and 34.5 DEG C of preheatings are stand-by；Matched group solvent processes；
The present invention also provides for a kind of measuring the medicine device to varoa mite toxicity.
The mensuration medicine of the present invention device to varoa mite toxicity, cultivates including Pupa Apis culture plate and Pupa Apis
Cup, it is characterised in that: if described Pupa Apis culture plate is to set dry hole, the size in described hole cultivates cup phase with Pupa Apis
Should, each hole can accommodate a Pupa Apis and cultivate cup；It is goblet that described Pupa Apis cultivates cup, and described Pupa Apis is cultivated
Filter paper is laid in the bottom of cup, and rim of a cup sets the ventilative transparent membrane for sealing.
In an embodiment of the invention, described Pupa Apis culture plate sets 24-48 and can accommodate Pupa Apis and cultivate
The hole of cup.
Further, assembly of the invention also includes providing cultivation temperature 34.5 DEG C, the relative humidity are
The incubator of 75%-95%.
The structure design of said apparatus, it is provided that the culture vessel of larva to Pupa Apis-Pupa Apis cultivation cup 2, meanwhile,
This container can also access varoa mite and have the medicine of different toxicity to varoa mite so that, detection more connects
Near-nature forest state, more life-like.Meanwhile, this device also facilitates the detection of multisample, utilizes the honeybee of porous
Pupa culture plate 1, makes one-time detection can carry out tens repetitions, is suitable for toxicity detection this needs large sample
The experiment of detection.
The inventive method utilizes said apparatus to detect, and its advantage is: (1) experimental condition is controlled；(2)
Matched group honeybee demodicid mite survival rate is high, and the time-to-live is long；(3) time saving and energy saving；(4) result is the most credible.
The method can be used for acaricide and measures the acute toxicity of varoa mite and chronic toxicity mensuration, kills for screening
Demodicid mite medicine provides easy believable assay method, is substantially shorter green and kills the research and development profit of honeybee demodicid mite medicine
With the cycle, existing important Research Significance has again wide market prospect.
Accompanying drawing explanation
Fig. 1 is that the present invention measures medicine and (puts into the side view of the device of varoa mite toxicity Pupa Apis and cultivate cup
24 hole Pupa Apis culture plate side views)；
Fig. 2 is Pupa Apis culture plate；
Fig. 3 is that Pupa Apis cultivates cup；
Fig. 4 is to put into bee larvae the Pupa Apis cultivation cup sealed；
Fig. 5 is larva culture plate；
Fig. 6 is that Apis grows each stage diagram.
Fig. 7 is toxicity test experiment, varoa mite to be measured and the growth of Apis.
Fig. 1-Fig. 5, in figure, 1 is Pupa Apis culture plate, and 2 cultivate cup for Pupa Apis, and 3 is breathing freely for sealing
Transparent membrane, 4 is filter paper, and 5 is Pupa Apis, and 6 is varoa mite.
In Fig. 6, caption is as follows:
| 1 day old larva (the first row is left) | 2 day old larva (in the first row) | 3 day old larva (the first row is right) |
| 4 day old larva (the second row is left) | 5 day old larva (in the second row) | 6 day old larva (the second row is right) |
| The larva (the third line is left) of capping | The larva (in the third line) of capping | The larva (the third line is right) of capping |
| Pupa (fourth line is left) | Pupa (in fourth line) | Pupa (fourth line is right) |
| Pupa (fifth line is left) | Pupa (in fifth line) | Pupa (fifth line is right) |
| Pupa (the 6th row is left) | Pupa (in the 6th row) | Turn into into honeybee (the 6th row is right) |
Detailed description of the invention
According to the present invention, it is provided that measure the medicine method to varoa mite toxicity, concrete, comprise the steps:
1) medicine to be measured is arranged according to process group, be configured to different concentration liquid to be measured with solvent, will treat
The healthy bee larvae body surface that measured quantity medicinal liquid is sprayed on gently, is then placed in one 34.5 DEG C by bee larvae
Pupa Apis after preheating is cultivated in cup；Matched group solvent processes；Lay the denseest bottom described Pupa Apis cultivation cup
The filter paper dried after degree medicine infiltration；
2) collecting varoa mite to be measured from the foster demodicid mite spleen prepared, the Pupa Apis of each process group and matched group is cultivated
One varoa mite of access in cup, and the rim of a cup of Pupa Apis cultivation cup is sealed with ventilative transparent membrane, Pupa Apis is trained
Foster cup is put in the hole of Pupa Apis culture plate, one, every hole, puts in incubator by Pupa Apis culture plate, and temperature is
34.5 DEG C, relative humidity is 75%；
3) add up the different disposal group varoa mite death toll of 48h or 72h, calculate mortality rate, analyze medicine
Acute toxicity to varoa mite；Or every day records the survival condition of test honeybee demodicid mite continuously, add up every honeybee demodicid mite
Time-to-live, analyze the medicine chronic toxicity to varoa mite；Wherein, the honeybee demodicid mite number/access of mortality rate=death
Honeybee demodicid mite number.
In described method, the bee larvae of described health is in capping larval phase.
In described method, the bee larvae of described health is the larva of Apis mellifera (Apis melifera).
In described method, the cultural method of the bee larvae of described health is as follows: with laying eggs, controller controls honeybee
Wang Yi opens and lays eggs on sky Nidus Vespae, discharges queen bee after 24h, after 3 days with Moving needle for bee-keeping larva is transferred to equipped with
In the larva culture plate being provided with some holes accommodating larva of 34.5 DEG C of preheatings of feedstuff, every hole moves into one
Bee larvae；Larva culture plate is placed in incubator, and temperature is 34.5 DEG C, relative humidity 95%, even
Continue and feed 6 days；Within 1st day, feed 10 μ L feedstuff A, within the 2nd day, feed 10 μ L feedstuff A, within the 3rd day, raise
Feed 20 μ L feedstuff B, within the 4th day, feed 30 μ L feedstuff C, within the 5th day, feed 40 μ L feedstuff C, the 6th day
Feed 50 μ L feedstuff C, obtain the bee larvae of health；Described feedstuff A by containing weight percentage is
The Lac regis apis of 44.25%, weight percentage be 5.3% glucose, weight percentage be the fruit of 5.3%
Sugar, weight percentage are yeast extract and water that weight percentage is 44.25% composition of 0.90%；
Described feedstuff B is by being 6.40% containing the Lac regis apis that weight percentage is 42.95%, weight percentage
Glucose, weight percentage be 6.40% fructose, weight percentage be 1.30% yeast extract
Thing and the water composition that weight percentage is 42.95%；Described feedstuff C by containing weight percentage is
The Lac regis apis of 50.00%, weight percentage be 9.00% glucose, weight percentage be 9.00%
Fructose, weight percentage are the yeast extract of 2.00% and water group that weight percentage is 30.00%
Become.
Concrete, the preparation method of the Pupa Apis cultivation cup after 34.5 DEG C of preheatings is as follows: medicine solvent to be measured is joined
Making different concentration liquid to be measured, dripped to by quantitative liquid medicine on the filter paper got ready, filter paper all soaks,
Then the filter paper of coating is placed in super-clean bench after solvent volatilization is dried, puts into Pupa Apis and cultivate cup
Bottom, each Pupa Apis cultivation one filter paper of cup, Pupa Apis is cultivated in the hole that cup is positioned over Pupa Apis culture plate,
Pupa Apis culture plate is placed in incubator, and 34.5 DEG C of preheatings are stand-by；Matched group solvent processes；
The method of the present invention employs the mensuration medicine device to varoa mite toxicity, mensuration of the present invention
The medicine device to varoa mite toxicity, cultivates cup 2, described Pupa Apis culture plate including Pupa Apis culture plate 1 and Pupa Apis
If being to set dry hole, it is corresponding that the size in described hole cultivates cup to Pupa Apis, and each hole can accommodate a Pupa Apis and cultivate cup；
Described Pupa Apis cultivate cup be goblet, described Pupa Apis cultivate cup bottom lay filter paper 4, rim of a cup set for
The ventilative transparent membrane 3 of sealing.
In an embodiment of the invention, described Pupa Apis culture plate sets 24-48 and can accommodate Pupa Apis and cultivate
The hole of cup.
Further, assembly of the invention also includes providing cultivation temperature 34.5 DEG C, the relative humidity are
The incubator of 75%-95%.
The structure design of said apparatus, it is provided that the culture vessel of larva to Pupa Apis-Pupa Apis cultivation cup, meanwhile,
This container can also access varoa mite and have the medicine of different toxicity to varoa mite so that, detection more connects
Near-nature forest state, more life-like.Meanwhile, this device also facilitates the detection of multisample, utilizes the honeybee of porous
Pupa culture plate 1, makes one-time detection can carry out tens repetitions, is suitable for toxicity detection this needs large sample
The experiment of detection.
Embodiment
Embodiment 1, the medicine assay method to the toxicity of varoa mite
Prepare before test: Apis mellifera (Apis melifera) bee colony (healthy group, infect honeybee demodicid mite group), training
Support case, super-clean bench, balance, magnetic stirring apparatus, mix agitator, Hygrothermograph, pipettor, super-clean bench
Heater, cold light source illuminates, controller of laying eggs, Moving needle for bee-keeping, tweezers, 48 hole larva culture plates, larva
Cultivate cup, honeybee demodicid mite catcher (culture dish with cover), shifting demodicid mite small brushes, Parafilm, Nidus Vespae incubation chamber,
Altex glove, mask, Lac regis apis, glucose, fructose, yeast extract, potassium sulfate, sodium chloride, double
Steam water, medicine to be measured.If no special instructions, medicine and equipment used by the present embodiment all can be from business ways
Footpath obtains.
1 prepares honeybee demodicid mite
Checking varoa mite parasitic rate in bee colony, the bee colony that host age preference rate is high is foster demodicid mite group, does not the most control demodicid mite.
When varoa mite virulence test starts, from foster demodicid mite group, take out the sub-spleen of new capping.Open lair one by one, collect
Varoa mite is standby.
2 raising honeybee larvas
Control queen bee with controller of laying eggs to lay eggs on an empty Nidus Vespae, after 24h, discharge queen bee, with moving after 3d
Larva is transferred to equipped with in the 48 warmed-up hole larva culture plates of man-made feeds (Fig. 5) by worm pin, every hole
Move into a larva.Culture plate is placed in incubator, and temperature is 34.5 DEG C, relative humidity 95%.Continuously
Feed 6 days, within the 1st day, feed 10 μ L feedstuff A, within the 2nd day, feed 10 μ L feedstuff A, within the 3rd day, feed
20 μ L feedstuff B, feed 30 μ L feedstuff C on the 4th day, within the 5th day, feed 40 μ L feedstuff C, within the 6th day, raise
Feeding 50 μ L feedstuff C, within the 7th day, be transferred to the present invention measures the medicine device to varoa mite toxicity.
Above-mentioned feed formula is as shown in table 1, and the percentage composition described in table 1 is weight percentage.
Table 1.
| Feedstuff A | Feedstuff B | Feedstuff C | |
| Lac regis apis | 44.25% | 42.95% | 50.00% |
| Glucose | 5.30% | 6.40% | 9.00% |
| Fructose | 5.30% | 6.40% | 9.00% |
| Yeast extract | 0.90% | 1.30% | 2.00% |
| Water | 44.25% | 42.95% | 30.00% |
Apis from ovum to going out room (emergence) 21 days.4th day (1 day old larva) starts to support at laboratory,
Be larva culture plate, the 10th day (capping larval phase), transfers to Pupa Apis culture plate, is administered and connects demodicid mite
Also it is this sky.Concrete operations see table 2.
Table 2.
3 dispensers
Utilize medicine of the present invention that the device of varoa mite toxicity is tested, Pupa Apis is cultivated cup (Fig. 3) and puts
In entering to 24 holes Pupa Apis culture plate (Fig. 2), every hole is put into a Pupa Apis and is cultivated cup (Fig. 1).Medicine to be measured
Thing is configured to variable concentrations according to test requirements document, is dripped to by quantitative liquid medicine on the little scraps of paper got ready, and the little scraps of paper are complete
Portion soaks, and the little scraps of paper of coating are placed in super-clean bench after solvent volatilization is dried, and puts into Pupa Apis and cultivates cup
In, each cultivation cup a piece of paper sheet, in Pupa Apis culture plate is placed into incubator (34.5 DEG C), preheating is stand-by.
Before larva goes to Pupa Apis culture plate, the body surface being sprayed on gently by medicinal liquid, then by larva from larva culture plate
Inside it is transferred in Pupa Apis culture plate.Matched group solvent processes.
4 connect demodicid mite
Collecting varoa mite from the foster demodicid mite spleen prepared, the Pupa Apis of each process group and matched group accesses in cultivating cup
One varoa mite, and seal culture hole (Fig. 3, Fig. 4) with ventilative transparent membrane, prevent the big honeybee accessed
Demodicid mite escapes from culture hole, it is simple to observe honeybee demodicid mite survival condition.Putting in incubator by Pupa Apis culture plate, temperature is
34.5 DEG C, relative humidity is 75%.
5 statistics honeybee demodicid mite mortality rates
Statistics 48h or 72h honeybee demodicid mite death toll, calculates mortality rate, analyzes the medicine acute toxicity to varoa mite.
Or every day records the survival condition of test honeybee demodicid mite continuously, add up the time-to-live of every honeybee demodicid mite, analyze medicine pair
The chronic toxicity of varoa mite.
The honeybee demodicid mite number of the honeybee demodicid mite number/access of mortality rate=death.
6, the coumafos (Coumaphos) toxicity test to varoa mite
The coumafos (Coumaphos) toxicity test to varoa mite is carried out according to above-mentioned steps 1-5, each
Process 3 repetitions, often repeat 12 varoa mites, as shown in table 2, D10 (i.e. capping larva the 1st day)
Chemicals treatment, is concurrently accessed honeybee demodicid mite, adds up honeybee demodicid mite mortality rate after 48h.Concrete outcome is as shown in table 3.
Table 3.
In table 3,1ppm=0.001 ‰, percentage by weight.
7, the coumafos (Coumaphos) chronic toxicity test to varoa mite
The coumafos (Coumaphos) chronic toxicity test to varoa mite is carried out according to above-mentioned steps 1-5,
In the Apis stage of development as shown in table 2, Apis grows the 10th day (i.e. capping larva the 1st day) and connects
Entering honeybee demodicid mite, the 21st day (Apis goes out room or emergence) checks mortality rate.Concrete outcome is as shown in table 4, its
The picture that middle honeybee demodicid mite survives on Apis body as it is shown in fig. 7, wherein, is that honeybee demodicid mite is honeybee from left first picture
Pupa is cultivated in cup, does not sees varoa mite, is Apis from left 2-4 pictures and honeybee demodicid mite is cultivated cup from Pupa Apis
Clap after taking out, on left 4th pictures, show the offspring of honeybee demodicid mite breeding.
Table 4.
In table 4,1ppm=0.001 ‰, percentage by weight.
Above-mentioned experiment shows, the advantage of the inventive method: (1) experimental condition is controlled；(2) matched group honeybee demodicid mite
Survival rate is high, and the time-to-live is long；(3) time saving and energy saving；(4) result is the most credible.
The method can be used for acaricide and measures the acute toxicity of varoa mite and chronic toxicity mensuration, kills for screening
Demodicid mite medicine provides easy believable assay method, is substantially shorter green and kills the research and development profit of honeybee demodicid mite medicine
With the cycle, existing important Research Significance has again wide market prospect.
Below it is only the preferred embodiment of the present invention, it is noted that for the ordinary skill of the art
For personnel, under the premise without departing from the principles of the invention, it is also possible to make some improvements and modifications, these
Improvements and modifications also should be regarded as protection scope of the present invention.
Claims (7)
1. measure a medicine method to varoa mite toxicity, comprise the steps:
1) medicine to be measured is arranged according to process group, be configured to different concentration liquid to be measured with solvent, will treat
The healthy bee larvae body surface that measured quantity medicinal liquid is sprayed on gently, is then placed in one 34.5 DEG C by bee larvae
Pupa Apis after preheating is cultivated in cup；Matched group solvent processes；Lay the denseest bottom described Pupa Apis cultivation cup
The filter paper dried after degree medicine infiltration；
2) collecting varoa mite to be measured from the foster demodicid mite spleen prepared, the Pupa Apis of each process group and matched group is cultivated
One varoa mite of access in cup, and the rim of a cup of Pupa Apis cultivation cup is sealed with ventilative transparent membrane, Pupa Apis is trained
Foster cup is put in the hole of Pupa Apis culture plate, one, every hole, is put into by Pupa Apis culture plate in incubator and cultivates, temperature
Degree is 34.5 DEG C, and relative humidity is 75%；
3) add up the different disposal group varoa mite death toll of different time sections, calculate mortality rate, analyze medicine
Acute toxicity to varoa mite；Or every day records the survival condition of test honeybee demodicid mite continuously, add up every honeybee demodicid mite
Time-to-live, analyze the medicine chronic toxicity to varoa mite；Wherein, the honeybee demodicid mite number/access of mortality rate=death
Honeybee demodicid mite number.
Method the most according to claim 1, it is characterised in that: the bee larvae of described health is in envelope
The larval phase of lid.
Method the most according to claim 1, it is characterised in that: the bee larvae of described health is west
The larva of Apis (Apis melifera).
Method the most according to claim 1, it is characterised in that: the training of the bee larvae of described health
Breeding method is as follows: controls queen bee with controller of laying eggs and lays eggs on an empty Nidus Vespae, discharges queen bee after 24h,
With Moving needle for bee-keeping, larva is transferred to being provided with of 34.5 DEG C of preheatings equipped with feedstuff after 3 days and some accommodates larva
Hole larva culture plate in, every hole move into a bee larvae；Larva culture plate is placed in incubator,
Temperature is 34.5 DEG C, and relative humidity 95% feeds 6 days continuously；Within 1st day, feed 10 μ L feedstuff A, the
Within 2 days, feed 10 μ L feedstuff A, within the 3rd day, feed 20 μ L feedstuff B, within the 4th day, feed 30 μ L feedstuff C,
Within 5th day, feed 40 μ L feedstuff C, within the 6th day, feed 50 μ L feedstuff C, obtain the bee larvae of health；
Described feedstuff A is the Fructus Vitis viniferae of 5.3% by the Lac regis apis that weight percentage is 44.25%, weight percentage
Sugar, weight percentage be 5.3% fructose, weight percentage be 0.90% yeast extract and weight
Amount percentage composition is the water composition of 44.25%；Described feedstuff B is the queen bee of 42.95% by weight percentage
Slurry, weight percentage be 6.40% glucose, weight percentage be 6.40% fructose, weight hundred
The yeast extract and the water that weight percentage is 42.95% that divide content to be 1.30% form；Described feedstuff C
By glucose, weight that the Lac regis apis that weight percentage is 50.00%, weight percentage are 9.00%
Percentage composition be 9.00% fructose, weight percentage be 2.00% yeast extract and weight percent contain
Amount is the water composition of 30.00%.
5. measure a medicine device to varoa mite toxicity, cultivate cup including Pupa Apis culture plate and Pupa Apis, its
Being characterised by: if described Pupa Apis culture plate is to set dry hole, it is corresponding, often that the size in described hole cultivates cup to Pupa Apis
Individual hole can accommodate a Pupa Apis and cultivate cup；It is goblet that described Pupa Apis cultivates cup, and described Pupa Apis cultivates the end of cup
Filter paper is laid in portion, and rim of a cup sets the ventilative transparent membrane for sealing.
Device the most according to claim 5, it is characterised in that: described Pupa Apis culture plate sets 24-48
Pupa Apis can be accommodated and cultivate the hole of cup.
Device the most according to claim 5, it is characterised in that: described device also includes providing cultivation
Temperature 34.5 DEG C, relative humidity are the incubator of 75%-95%, during use, and described Pupa Apis culture plate and honeybee
Pupa is cultivated cup and is placed in incubator.
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