CN106719786A - A kind of application of red fire ant contact insecticide and ethiprole in killing red ant - Google Patents
A kind of application of red fire ant contact insecticide and ethiprole in killing red ant Download PDFInfo
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- CN106719786A CN106719786A CN201710009601.2A CN201710009601A CN106719786A CN 106719786 A CN106719786 A CN 106719786A CN 201710009601 A CN201710009601 A CN 201710009601A CN 106719786 A CN106719786 A CN 106719786A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N57/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds
- A01N57/10—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds having phosphorus-to-oxygen bonds or phosphorus-to-sulfur bonds
- A01N57/14—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds having phosphorus-to-oxygen bonds or phosphorus-to-sulfur bonds containing aromatic radicals
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/002—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing a foodstuff as carrier or diluent, i.e. baits
- A01N25/006—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing a foodstuff as carrier or diluent, i.e. baits insecticidal
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N47/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid
- A01N47/02—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid the carbon atom having no bond to a nitrogen atom
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N57/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds
- A01N57/10—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds having phosphorus-to-oxygen bonds or phosphorus-to-sulfur bonds
- A01N57/16—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds having phosphorus-to-oxygen bonds or phosphorus-to-sulfur bonds containing heterocyclic radicals
Abstract
The invention discloses a kind of application of red fire ant contact insecticide and ethiprole in killing red ant.A kind of red fire ant contact insecticide, its active ingredient is ethiprole, phoxim and/or chlopyrifos.Another object of the present invention is to provide application of the ethiprole in killing red ant.Described application, it is that the ant bait formulation of killing containing ethiprole is taken food by red fire ant worker ant, after bait formulation is brought into nest by red fire ant, frequent activities and trophallaxis feeding behavior by worker ant in nest, medicament is delivered to worker ant in nest, young ant and queen, causes full nest red fire ant to be poisoned to death.The present invention is prepared simply, and easy to use, insect killing effect is good.
Description
Technical field
The present invention relates to insecticide, and in particular to a kind of red fire ant contact insecticide and ethiprole are in killing red ant
Using.
Background technology
Red fire ant (S.invicta) is the social insect of most invasive, is classified as in the world and most invaded
One of property and destructive hundred kinds of Exotic pests.Red fire ant feeding habits are complicated, habit is violent, breeding is rapid, competitiveness is strong, energy
Causing the omnivorousness and aggressiveness of heavy economic losses, social danger and ambient influnence, red fire ant makes it be not only a kind of economy
Insect, also threatens public health.Chemical control methods are still preventing and treating red fire ant effective measures, but still can not be eradicated flourishing
Ant.And red fire ant is controlled with chemical agent for a long time, unexpected bounce-back can be caused.
Ethiprole (trade name：Frontline) english common name is fipronil, be Luo Na-Rhone-Poulenc of former France in
The phenyl pyrazoles New-type wide-spectrum insecticide of exploitation in 1981, chemical name：(±) -5- amino -1- (2,6- bis- chloro- 4,4,
4- fluoroforms phenyl) -4- trifluoromethyl sulfinyl -3- cyano pyrazoles.Ethiprole can be combined with GABA receptor, be had
The chloride channel of the GABA regulation of effect obstruction insect CNS, interference insect central nervous system causes
Insect N＆M is overexcited until dead.Used as the insecticide of control crop pest, ethiprole activity is high, insecticidal spectrum
Extensively, have tag, stomach toxicity and systemic action, to Lepidoptera, Homoptera, Semiptera, Thysanoptera and Orthoptera, coleoptera and Hymenoptera
Various pests have excellent preventive effect.
Ethiprole is due to that with unique mechanism of action, can efficiently control to organophosphor, carbamate, pyrethroid
Ester pesticide produces the insect of resistance.In recent years, it has been found that some insects produce a certain degree of resistance to ethiprole.
Kolaczinski etc. (2001) is tested the anopheles of 5 strains with WHO set reagents, finds two of which strain to fluorine
Worm nitrile is resistant.Ethiprole is metabolized the generation sulfone that toxicity is suitable with ethiprole or toxicity is higher in animal, plant, environment
Thing or sulfoxide compound.In animal body by absorbing, metabolism, drain and acted on with hydrolysis, photodissociation etc., be distributed widely in animal
Tissue in, particularly content is higher in adipose tissue.Main discharge approach is by excrement.Concrete ways are as follows：
Ethiprole metabolic pathway in animal body.
Chlopyrifos (chlorpyrifos) be it is a kind of efficiently, the organophosphorus insecticide of wide spectrum, with tagging, stomach toxicity and stifling
Effect, can be used to prevent and treat the insect chlopyrifos pair such as snout moth's larva, leaf folder, mythimna separata, scale insect, aphid, bollworm, leafhopper and red spider
Nervous system has toxic action, and the oxidation chlopyrifos than 3000 times of itself strong toxicity can be converted in animal body.
Primary metabolite in water, air, soil, plant and animal body is higher than chlopyrifos toxicity to chicken placenta 2-3 times.And chlopyrifos
Cooperative effect is there may exist with TCP (3,5,6- tri- chloro- 2- pyridols).Chlorpyrifos pesticide is aoxidized with Cytochrome P450 list
Enzyme induction research focuses mostly in fish, the mankind and mouse (Wheelock and Eder, 2005；Cui et al.,2008；
Lee, 2009), in insect, and the research to FscP is still few.Recently, in Rong Rong etc. (2013) poison
Dead tick induction Asiatic migrotory locust, as a result shows, CYP4G62 can significantly be induced by high dose chlopyrifos, and CYP9AQ1 is processed through chlopyrifos
Afterwards, significantly induced in low, medium and high concentration.
The content of the invention
In order to solve the deficiencies in the prior art, killed the invention provides a kind of red fire ant contact insecticide and ethiprole
Application in red fire ant.
The technical scheme is that：A kind of red fire ant contact insecticide, its active ingredient be ethiprole, phoxim and/or
Chlopyrifos.
Further improvement of the present invention includes：
Its effectively become be concentration be 0.15-0.36 μ g/ml phoxim or 0.26-0.55 μ g/ml ethiprole or
The chlopyrifos of 0.34-0.64 μ g/ml.
Described red fire ant contact insecticide is that active ingredient is dissolved in acetone to be obtained.
Its active ingredient is that concentration is not less than 15.168 μ g/ml ethiproles.
Another object of the present invention is to provide application of the ethiprole in killing red ant.
Described application, is that the ant bait formulation of killing containing ethiprole is taken food by red fire ant worker ant, when bait formulation is brought into by red fire ant
After in nest, medicament is delivered to worker ant in nest, young ant and ant for frequent activities and trophallaxis feeding behavior by worker ant in nest
Afterwards, full nest red fire ant is caused to be poisoned to death.
The present invention is prepared simply, and easy to use, insect killing effect is good.
Brief description of the drawings
Fig. 1 is BSA protein content standard curves.
Fig. 2 are the absorbances of various concentrations p-nitrophenol
Specific embodiment
The present invention is elaborated below in conjunction with the accompanying drawings.
Embodiment 1
The prevention and controls of red fire ant are a lot, wherein Techniques For Chemical Control be it is most effective, most deep, be most widely used it is anti-
Control method.Broadcasting sowing for bait formulation is maximally effective and holding effect the mode of red fire ant large area control, fills nest (mound drench)
Be with when, effectively, one of effective ways of fast preventing red fire ant.The application selects 7 kinds of different types of insecticides to flourishing
The Contact toxicity of ant is determined, and specifies different agents right to the Virulence Difference of red fire ant and different brackets red fire ant (queen and worker ant)
The sensitivity difference of medicament.
Test material
Test worm and material
Guangdong Province Guangzhou Zengcheng and Campus of South China Agricultural University are picked up from for examination red fire ant.With reference to (2000) such as Kuriachan
Method, red fire ant ant colony is adopted back with big Plastic Drum from field, places 1-2d, after ant colony establishes new group in bucket, will
Running water is slowly dropped into bucket, makes red fire ant automatic upper shifting, emersion from group.Ant colony is transferred into basin mouthful with the dredger of 40 mesh to apply
In with the plastic tub (7.0L) of talcum powder, (25 DEG C of room temperature, RH 65%-80%) is manually supplied after feed is raised one week indoors
(Zeng Xinnian etc., 2006) is used in experiment.Worker ant, soldier ant, the criterion reference for having wing aner, having wing gyne and queen and young ant were once
Tinkling of pieces of jade etc. (2005) method.
Reagent agent
Medicament
96% chlopyrifos original powder Chlorpyrifos；
95% effective cypermethrin active compound Beta-cypermethrin；
91% phoxim emulsifiable concentrate Phoxim；
96% chlopyrifos Chlorpyrifos；
97% imidacloprid Imidacloprid；
90% ethiprole Fipronil；
95% gamma cyhalothrin Ambda-cyhalothrin；
97.2% pyridine worm miaow Acetamiprid；
96.2% pleocidin Spinosad；
93.7% AVM Abamectin.
Biochemical reagents
Coomassie brilliant G-250, Coomassie Brilliant Blue G250；
Bovine serum albumin (BSA), Bovine serum albumin；
Analytical reagent and organic solvent
Disodium hydrogen phosphate, Disodium hydrogen phospHate are analyzed pure；
Sodium dihydrogen phosphate, Sodium dihydrogen phospHate are analyzed pure；
NaOH, Sodium hydroxide are analyzed pure；
Acetic acid, Acetate is analyzed pure；
Sodium acetate, Sodium acetate are analyzed pure；
Sodium carbonate, Sodium carbonate are analyzed pure；
Sodium acid carbonate, Sodium blcarbonate are analyzed pure；
Hydrochloric acid, Hydrochlori cacid are analyzed pure；
Acetone, Acetone is analyzed pure.
Key instrument equipment
Illumination box (LRH-300-GH)；
Electronic balance (BS2190S, accuracy 0.1mg)；
Ultraviolet specrophotometer (UV-9100 types)；
Micro intravenous drip instrument (HMA 0807140)；
High speed freezing centrifuge (M IKRO 22R types)；
Micro syringe (1-5 μ l)；
Water-bath (HH.S21-4 types)；
HOSHIZAKI ice machines (FM-120EE)；
KH-500 ultrasonic cleaners.
Test method
Contact toxicity is determined
Medicine embrane method:Active compound is dissolved in proper amount of acetone a series of medicament for being made into concentration, 1ml is pipetted after preliminary experiment
In acetone liquid to bore, bottom footpath, the respectively disposable plastic cup of 6.6,4.4 and 6.8cm high, the coating area of cup is
72.85cm2, rotating cup makes liquid be spread evenly across inwall, after acetone volatilization is dry, is inverted cup, smears appropriate in rim of a cup
Talcum powder, selects 10 queens or 50 worker ants in cup at random.Each concentration sets 3 repetitions, and blank is acetone.Observation
Dead ant number after record 24h and 48h, calculates the death rate.Virulence regression equation is asked for by niqueMin method and least square method, is counted
Calculate the lethal concentration of 50 (LC50) value.
Drop method：Active compound is dissolved in proper amount of acetone a series of medicament for being made into concentration, queen is selected, is had wing gyne each
10, worker ant 30, liquids of the 1 μ l through the above-mentioned concentration after preliminary experiment is pipetted with micro intravenous drip instrument, and drop is in thoracic dorsal before test worm
Plate, is placed in disposable plastic cup, and appropriate talcum powder is smeared in rim of a cup, and with the sucrose water of cotton pellet supply 10%, each is dense
Degree sets 3 repetitions, and acetone drop is used in control.Observed and recorded 24h death borer populations, the death rate is calculated by (2.1) and (2.2) respectively
(%) and corrected mortality (%).With SPSS, (software statistics treatment, obtains virulence regression equation and LC to result statistics50And LC90。
Statistical analysis technique
Biometric data is analyzed using Software of Data Statistics POLO, soft using Microsoft EXCEL2010
Part arranges and data is carried out with statistical analysis (one-way ANOVA and Tukey ' s with GraphPad InStat softwares
test,P<0.05)。
Result and analysis
Insecticide is planted to act on red fire ant worker ant Contact toxicity
Under indoor conditions, 7 kinds of insecticides are determined using medicine embrane method red fire ant worker ant 24h Contact toxicity result of the tests are shown in Table
2-1.Result shows, LC of the phoxim to worker ant50And LC90Respectively：0.15 and 0.36 μ g/ml, gamma cyhalothrin is to work
The LC of ant50And LC90Respectively：0.24 and 0.38 μ g/ml；LC of the ethiprole to worker ant50And LC90Respectively：0.26 and 0.55 μ
g/ml；LC of the chlopyrifos to worker ant50And LC90Respectively：0.34 and 0.64 μ g/ml；LC of the imidacloprid to worker ant50And LC90Respectively
For：663.46 and 877.43 μ g/ml；LC of the pleocidin to worker ant50And LC90Respectively：1068.90 and 1351.11 μ g/ml.
This shows that red fire ant worker ant is more sensitive to most insecticide, tries have 5 kinds of LC of pesticide treatments 24h in 7 kinds of medicaments50Value is low
In 0.5 μ g/ml；Though the above-mentioned four kinds of agricultural chemicals of pleocidin Contact toxicity discord, relatively preferable, only the poison of tagging of AVM
Power is most weak.Ethiprole, phoxim and chlopyrifos also have good tactile cytotoxicity.
Contact toxicity (1.6mg/) of the 7 kinds of insecticides of table 2-1. to red fire ant worker ant
χ2(df)：Chi-square value (free degree).
Ethiprole is to worker ant and the Contact toxicity of queen
Under indoor conditions, using topical application method ethiprole red fire ant worker ant and the 24h Contact toxicity result of the tests of queen
(table 2-2) is visible, and ethiprole is to worker ant and the LC of queen50Respectively 5.12 and 15.168 μ g/ml.Ethiprole is to red fire ant worker ant
Individual with queen very big to the sensitivity differences of medicament, virulence can differ 2-3 times or so.
Table 2-2. ethiproles are to red fire ant worker ant and the Contact toxicity of queen
χ2(df)：Chi-square value (free degree).
Brief summary
Under indoor conditions, 7 kinds of insecticides are determined to red fire ant worker ant 24h Contact toxicities using medicine embrane method.Result shows, red
Fiery ant worker ant is more sensitive to most insecticide, tries have 5 kinds of LC of pesticide treatments 24h in 7 kinds of medicaments50Value is less than 0.5 μ g/
ml；Though pleocidin Contact toxicity not four kinds of agricultural chemicals as described above, effect is relatively preferable, only the Contact toxicity of AVM is most
It is weak.Ethiprole, phoxim and chlopyrifos also have good tactile cytotoxicity.Simultaneously using topical application method ethiprole to flourishing
Ant worker ant is tested with the 24h Contact toxicities of queen, as a result shows, ethiprole is to worker ant and the LC of queen50Respectively 4.604 Hes
15.168μg/ml.Ethiprole is individual to red fire ant worker ant and queen very big to the sensitivity differences of medicament, and virulence can differ 2-
3 times.Queen shows certain drug resistance compared with worker ant.
Embodiment 2
Cytochrome P450 (Cytochrome P450, abbreviation P450) is the core of mixed-function oxidases, whole
Terminal oxidized effect (Qiu Lihong etc., 1999 is played in the oxidative metabolism of individual enzyme system；Isin and Guengerich,2007).
P450 can in involved in insect body various allogenic materials and endogenous substance metabolism.Cytochrome P450 determination of activity is conventional following
Several method：The de- ethyl methods of 1.O-：By CYP1A ethoxyresorufin O-deethylase (7-ethoxycoumarin O-
Deacthylase, ECOD) activity characterization cytochrome P 450 Enzyme activity.CYP1A (7-
Ethoxycoumarin umbelliferone (7-hydroxycoumarin)) is changed into after de-ethylation, and fluorescent value can be produced afterwards
Changing (excitation wavelength 368nm, launch wavelength 456nm), using sepectrophotofluorometer, surveys umbelliferone generation volume production
The change of raw fluorescent value characterizes activity (Ullrich, 1972 of Cytochrome P450；Aitio,1978).2.O- demethylation methods：
Using paranitroanisole (p-nitroanisole) p-nitrophenol is generated under the O- demethylations of Cytochrome P450
(p-nitrophenol) p- under spectrophotometer (Fredand and Hodgson, 1980) detection alkalescence condition can, be used
Absworption peaks of the nitrophenol at 405nm.Ai its people's foundation ethyl acetate/n-hexane leads to as terminator and extractant
Gradient Elution is crossed, the growing amount of p-nitrophenol is analyzed, sensitivity and the resolution of test is significantly improved
(Ai its people etc., 2009)；3. drinox epoxidation process (aldrinepoxidation, AE)：It is turned into Cytochrome P450 epoxy
Dieldrite is generated with lower drinox, then again by the amount of gas chromatographic detection dieldrite, so as to determine Cytochrome P450
Activity (Hammock et al., 1975；Yu et al.,1982).
The application determines the enzymatic activity of red fire ant P450 using P450O- demethylations method, with paranitroanisole (p-
Nitroanisole be) substrate, probe into red fire ant worker ant and queen P450 enzyme activity sex differernce with insecticide to red fire ant cell
The inducing action of cytochrome p 450 O- demethylating activities.Specify that xenobiontics are induced red fire ant worker ant P450 enzymatic activitys when
Between effect and dosage effect so that be further research red fire ant mechanism of drug resistance and red fire ant science administer theory is provided
Foundation.
Materials and methods
Selected insect source and raising
With embodiment 1.
Chemical reagent
NADPH (98% purity), bovine serum albumin(BSA) (BSA)；Tris-base, dithiothreitol (DTT) (DTT, 99% purity)；
Phenylthiourea (PTU)；Phenylthiourea (PTU)；Phenylmethylsulfonyl fluoride (PMSF)；Ethylenediamine tetra-acetic acid (EDTA), 7- ethoxy basic notes
Legumin (7-ethoxycoumarin), umbelliferone (7-hydroxycoumarin)；Paranitroanisole, p-
nitroanisole；P-nitrophenol p-nitrophenol；Coomassie brilliant G-250；Ethylenediamine tetra-acetic acid (ethylene
Diamine tetraacetic acid, EDTA), glycine (Gly), trichloroacetic acid (TCA), glycerine, 85% phosphoric acid, acetone,
NaH2PO4And Na2HPO4。
Laboratory apparatus
5417C/R types table-type high-speed refrigerated centrifuge (Eppendorf, Germany)；LS-55 sepectrophotofluorometers
(Perkin Elmer, USA)；Bio-40 spectrophotometries instrument (Perkin Elmer, USA)；Sartorius
2004M electronic balances (Opton, 13Germany)；SPH-100B type shaking tables；Water-bath.
Experimental technique
The neat red fire ant some heads of worker ant adult of development are chosen, ethiprole LC is used respectively50(0.25 μ g/ml), chlopyrifos
LC50(0.34 μ g/ml) treatment, with acetone treatment as control.It is put into the disposal plastic cup for scribbling medicine film, is placed in environment bar
Part is 26 ± 2 DEG C of temperature, relative humidity 50%-60%, photoperiod 16h：8h(L：D in greenhouse), respectively after treatment 24,
48th, 72 and 96h samplings, are stored in -80 DEG C of refrigerators for determining P-NA O- demethylating activities.
P450 enzyme assays
The preparation of enzyme liquid
Method with reference to rainbow etc. (2002) is simultaneously modified slightly, and it is some to take red fire ant, is added in 60/ml ratios certain
0.1mol/L, pH 7.5 of amount phosphate buffer (EDTA containing 1mmol/L, 1mmol/L PMSF, 1mmol/L DTT and
10% glycerine) homogenate, 4 DEG C, 10800rpm centrifugation 20min are de- for P-NA O- with absorbent cotton filtering supernatant as enzyme liquid
The measure of methyl activity and protein content.
P-NA O- demethylating activities are determined
With reference to Yu etc. (1992) and improved, reacted：Take 375 μ l 2mM PNA, 30 μ l 9.6mM NADPH, and 345 μ
The thick enzyme source mixing of l, 34 DEG C, exposed to water-bath 30min in air, add the reaction solution of 200 μ l, at 405nm in ELISA Plate
Mensuration absorbance.The result of generation is calculated with p-nitrophenol amount, enzyme activity unit is：μmol/pro(mg)/30min.
P-nitrophenol standard curve making
1、10-5Mol/L p-nitrophenyl phenol solutions：Accurately 0.03478g p-nitrophenols are weighed to be dissolved in the ethanol of 2.5ml,
Treat that the used time dilutes 10000 times with ethanol again.
2nd, 0.5mol/L NaOH solutions：Accurately 0.1g NaOH are weighed to be dissolved completely in the distilled water of 5ml.
3rd, by the standard of 0,20,40,80,120,160,200 μ l by 10-5Mol/L p-nitrophenyl phenol solutions are added separately to
In ELISA Plate hole, less than 250 μ l, with 0.5mol/L NaOH solution filling-in.Each concentration is at least repeated 3 times.
4th, its absorbance (OD values) is determined with ELIASA at A405, draws standard curve.
Determining the protein quantity
With reference to the method for the Coomassie brilliant G-250 of Bradford (1976), albumen is made with bovine serum albumin(BSA) (BSA)
Matter standard curve.Test tube is divided into 7 groups, every group sets 3 repetitions, sets up the reaction system (such as table 3-1) of 3ml.
Table 3-1. protein standard curves are determined
Table 3-1.The method of protein standard curve
2.5ml G-250+0.5ml determine liquid liquid and mix, and room temperature places 5min, are determined with spectrophotometry instrument
Light absorption value at 595nm.With OD values as abscissa, BSA concentration makes protein standard curve for ordinate.Enzyme liquid albumen is determined to contain
During amount, original enzyme liquid is diluted into 30-50 times, the same reaction system using 3ml, wherein enzyme liquid 0.5ml, G-250 2.5ml, room temperature
Light absorption value at 595nm is determined after reaction 5min, the protein content in enzyme liquid is calculated according to protein standard curve.
Data statistic analysis
Red fire ant paranitroanisole (p-nitroanisole) is calculated in Cytochrome P450 according to Rate activity formula
The activity of O- demethylations：
Rate activity (μm ol/min/mgPro.)=(OD × V is always/(T × Pro. × V enzymes)
Wherein, OD is light absorption value of the product p-nitrophenol (p-nitrophenol) (μm ol/ml) at 405nm；
V is always reaction cumulative volume (ml)；T is the reaction time (min)；Pro. it is original enzyme liquid protein content (mg/ml)；V is reaction enzyme liquid
Volume (ml).
Result and analysis
Protein content standard curve
Measurement result is as shown in Figure 1.Bovine serum albumin content (μ g/ml) is independent variable (X), with OD595It is dependent variable to be worth
(Y), fitting protein content calibration curve equation is：Y=0.0067x+0.1154, R2=0.9948, to calibration curve equation
Regression relation carry out t inspections, show as significant difference.
The standard curve of p-nitrophenol
Measurement result is as shown in Figure 2.Independent variable (X), OD are made with the concentration value (nmol/L) of p-nitrophenol400Value make because
Variable (Y), be fitted p-nitrophenol calibration curve equation be：Y=3.18 × 10-2+1.41×10-2X, R2=0.9996, it is right
The regression relation of calibration curve equation carries out t inspections, shows as significant difference.
Different brackets red fire ant (queen and worker ant) Cytochrome P450 O- hepatic Microsomal Aniline Hydroxylases
Red fire ant different brackets (queen and worker ant) Cytochrome P450 O- demethylase measurement results are shown in Table 3-2., flourishing
Ant queen Cytochrome P450 O- hepatic Microsomal Aniline Hydroxylases are less than worker ant and significant difference, and its Rate activity is respectively 3.23 and 5.35 μ
Mol/mg/pro/30min, differs 1.66 times.
Table 3-2. different brackets red fire ant (queen and worker ant) Cytochrome P450 O- hepatic Microsomal Aniline Hydroxylases are determined
Table 3-2.The specific activities of cytochrome P450of different
castes of S.invicta
Note：Numerical value is average value ± standard error (μm ol/mg/30min protein) in table, different lowercases in same column
Different disposal significant difference (ANOVA, P ＜ 0.05) is represented, numerical value is the ratio of worker ant and queen group in bracket
Note:The results are shown as mean±S.E(μmol/mg/30min protein)；
different lowercase letters in the same column mean significant difference
(ANOVA, P ＜ 0.05), data in bracket denote fold between workers/queens.
Ethiprole and chlopyrifos are acted on red fire ant P450 activity inducements
Using medicine embrane method ethiprole LC50With chlopyrifos LC50Red fire ant worker ant, the cytochromes of measure are processed respectively
P450 Activity Results are shown in Table 3-3..Red fire ant worker ant P450 Activity Results show, during 24h, at ethiprole treatment group and chlopyrifos
Reason group P450 activity is significantly higher than control group, respectively the 1.97 of control group times and 1.71 times；During 48h, two treatment group P450 live
Property substantially reaches peak, respectively the 3.00 of control group times and 1.96 times；During 72h, two treatment group P450 activity is reduced, poison
Dead tick treatment group reduction becomes apparent, respectively the 2.43 of control group times and 1.27 times.During 96h, two treatment group P450 activity is still
Be higher than so control group, respectively the 2.26 of control group times and 1.12 times, chlopyrifos treatment group P450 it is active substantially with control group phase
It is flat.
Table 3-3. ethiproles LC50(0.05 μ g/ml) and chlopyrifos LC50Red fire ant (1.25mg/ after (0.15 μ g/ml) reason
Head) P450 activity
Note：Numerical value is average value ± standard error (μm ol/mg/30min protein) in table, same time point difference small letter
Letter represents different disposal significant difference (ANOVA, P ＜ 0.05), and numerical value is same time point treatment group and control group in bracket
Ratio
Brief summary
Red fire ant different brackets (queen and worker ant) Cytochrome P450 O- demethylase measurement results show, red fire ant ant
Cytochrome P450 O- hepatic Microsomal Aniline Hydroxylases are less than worker ant and significant difference afterwards, and its Rate activity is respectively 3.23 μm of ol/mg/pro/
30min and 5.35 μm of ol/mg/pro/30min, differs 1.66 times.Probably due to the difference of red fire ant queen and worker ant function, ant
It is afterwards the center of whole ant nest, is responsible for regulation and control ant colony population quantity, sex ratio etc., the outer activity of ant nest of seldom going out, and worker ant
Often go out to be engaged in the activity of looking for food etc., it is easy to touch xenobiontics and raise metabolic detoxification enzyme content.Show thin
Born of the same parents' cytochrome p 450 is in worker ant to the importance of drug detoxication metabolic function.
Using medicine embrane method ethiprole LC50With chlopyrifos LC50Red fire ant worker ant, the cytochromes of measure are processed respectively
P450 activity displays, during 24h, ethiprole treatment group and chlopyrifos treatment group P450 activity are significantly higher than control group, respectively right
According to 1.97 times and 1.71 times of group；During 48h, two treatment group P450 activity substantially reaches peak, respectively the 3.00 of control group
Again with 1.96 times；During 72h, two treatment group P450 activity reduces special chlopyrifos treatment group and is decreased obviously, respectively control group
2.43 times and 1.27 times.During 96h, two treatment group P450 activity be still above control group, respectively the 2.26 of control group times and
1.12 times, chlopyrifos treatment group P450 activity is substantially equal with control group.From entirety, relative to chlopyrifos, ethiprole is to red
Fiery ant has and preferably tags and inducing action, and with persistence.Ethiprole is a kind of fluorine-containing pyrazoles insecticides, activity height,
Insecticidal spectrum is wide, and ethiprole is set to a kind of medicament that organic phosphates can be replaced to prevent and treat ant for United States environmental protection agency
(U.S.EPA,2000).This laboratory research simultaneously shows that ethiprole kills ant bait formulation has excellent stomach poison function to red fire ant,
Worker ant takes food red fire ant, after bait formulation is brought into nest by red fire ant, frequent activities and trophallaxis feeding row by worker ant in nest
For, medicament is delivered to worker ant in nest, young ant and queen, cause full nest red fire ant to be poisoned to death, reach killing red ant ant
The purpose of nest.
General principle of the invention and principal character and advantages of the present invention has been shown and described above.The technology of the industry
Personnel it should be appreciated that the present invention is not limited to the above embodiments, simply explanation described in above-described embodiment and specification this
The principle of invention, without departing from the spirit and scope of the present invention, various changes and modifications of the present invention are possible, these changes
Change and improvement all fall within the protetion scope of the claimed invention.The claimed scope of the invention by appending claims and its
Equivalent thereof.
Claims (6)
1. a kind of red fire ant contact insecticide, it is characterised in that its active ingredient is ethiprole, phoxim and/or chlopyrifos.
2. a kind of red fire ant contact insecticide according to claim 1, it is characterised in that it is that concentration is that it effectively becomes
The chlopyrifos of the phoxim of 0.15-0.36 μ g/ml or the ethiprole of 0.26-0.55 μ g/ml or 0.34-0.64 μ g/ml.
3. a kind of red fire ant contact insecticide according to claim 1, it is characterised in that active ingredient is dissolved in acetone
In be obtained.
4. a kind of red fire ant contact insecticide according to claim 1, it is characterised in that its active ingredient is not low concentration
In 15.168 μ g/ml ethiproles.
5. application of the ethiprole in killing red ant.
6. application according to claim 4, it is characterised in that the ant bait formulation that kills containing ethiprole is taken by red fire ant worker ant
Food, after bait formulation is brought into nest by red fire ant, frequent activities and trophallaxis feeding behavior by worker ant in nest transmit medicament
Worker ant, young ant and queen in nest, cause full nest red fire ant to be poisoned to death.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101069513A (en) * | 2007-04-30 | 2007-11-14 | 珠海经济特区瑞农植保技术有限公司 | Solenopsis invicta buren touch-killing powder agent |
| CN101601395A (en) * | 2009-03-02 | 2009-12-16 | 深圳市捷立克卫生服务有限公司 | The bait for luring and killing red fire ants of improvement |
| CN101669528A (en) * | 2009-09-17 | 2010-03-17 | 惠州市南天生物科技有限公司 | Solenopsis invicta buren bait |
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2017
- 2017-01-06 CN CN201710009601.2A patent/CN106719786A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101069513A (en) * | 2007-04-30 | 2007-11-14 | 珠海经济特区瑞农植保技术有限公司 | Solenopsis invicta buren touch-killing powder agent |
| CN101601395A (en) * | 2009-03-02 | 2009-12-16 | 深圳市捷立克卫生服务有限公司 | The bait for luring and killing red fire ants of improvement |
| CN101669528A (en) * | 2009-09-17 | 2010-03-17 | 惠州市南天生物科技有限公司 | Solenopsis invicta buren bait |
Non-Patent Citations (1)
| Title |
|---|
| 黄田福 等: "15种杀虫剂对红火蚁工蚁的触杀活性研究", 《华南农业大学学报》 * |
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