KR20170093413A - Effect of chlorine dioxide on disruption of insect acetylcholinesterase and its insecticidal activity - Google Patents
Effect of chlorine dioxide on disruption of insect acetylcholinesterase and its insecticidal activity Download PDFInfo
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- A—HUMAN NECESSITIES
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- 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
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Abstract
Description
The present invention relates to an insecticidal composition for increasing the activity of raising an acetylcholine ester containing chlorine dioxide, and more particularly to an insecticidal composition for increasing the acetylcholine ester raising activity by chlorine dioxide treatment.
Chlorine dioxide (ClO 2 ) is a disinfectant and bleaching agent with high oxidizing power and is being used for the sterilization of various microorganisms during the production of drinking water. Chlorine dioxide also has an excellent disinfecting effect on hospital bacteria, oral polluted bacteria, drinking water contaminated bacteria and common dish polluted bacteria that cause human food contamination. For bacteria that are particularly resistant to several antibiotics, chlorine dioxide exhibits excellent antibiotic ability compared to conventional sodium hypochlorite (NaClO). In addition, chlorine dioxide is known to cause virus inactivation against a variety of human pathogenic viruses including enterovirus 71 (EV71), which causes not only bacteria but also water-borne diseases.
As to the above chlorine dioxide, a function as a pesticide fumigant for hygienic pests that harm the living environment of a person by converting chlorine dioxide into a gaseous state and low pests that cause damage to stored cereals has been reported. Bleeds in hospital facilities ( Cimex lectularius, Cimex complete control in exposure of hemipterus) a relatively high concentration (with respect to chlorine dioxide exposure of about 1,000 ppm) showed a fast-acting control effect, low grain hwaranggok moth (relatively low concentrations (200 ppm) for Plodia interpunctella), which is applied to for Effect. Also, a false rice thief ( Tribolium castaneum ), chlorine dioxide fumigation showed high insecticidal activity.
The functional groups of chlorine dioxide showing these antibacterial and insecticidal effects have not yet been clarified. It is generally expected that the compounds will exhibit antibacterial and insecticidal activity due to their high oxidizing power. That is, exposure to chlorine dioxide oxidizes aromatic amino acids to induce denaturation of proteins, thereby losing protein function. It is also known that chlorine dioxide directly acts to change the nucleic acid of DNA or RNA. Recently, chlorine dioxide has been reported to cause a high mortality of treated insects by causing a large amount of active oxygen in treated insects, and active oxygen is known to affect relatively diverse biomolecules. However, The molecular end point of active oxygen has not been elucidated.
On the other hand, acetylcholine esterase (AChE) is an enzyme that plays a role in decomposing acetylcholine, a neurotransmitter, in the central nervous system of insects. It is known that the inhibition of AChE activity is associated with insecticidal activity (Devonshire, A. L., 1975, Biochem. J. 149, 463-469). The molecular end point of organophosphorus and carbamate insecticides is AChE, and inhibition of AChE induces hyperactivity of the nervous system, leading to death of the insect. In other words, insecticides against insect pests are known to inhibit the AChE enzyme, leading to the lethal phase of insect pests.
The inventors of the present invention have conducted research on insecticides of low-pest insects and found that when the increase of acetylcholine ester is increased by chlorine dioxide, insecticidal power is increased, thus completing the present invention.
Accordingly, an object of the present invention is to provide a composition for increasing the acetylcholine ester raising activity comprising chlorine dioxide.
However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.
In order to achieve the above object, the present invention provides a composition for increasing the acetylcholine ester raising activity of a pest including chlorine dioxide.
In one embodiment of the present invention, the concentration of chlorine dioxide in the composition is 8 to 1000 ppm.
In another embodiment of the present invention, the insect pest is Plodia interpunctella .
The present invention also provides an insecticidal composition comprising the composition for increasing activity.
The present invention also provides a method for increasing the acetylcholine ester raise activity of a pest comprising treating chlorine dioxide.
In one embodiment of the present invention, the chlorine dioxide treatment is carried out by intramuscular injection or fumigation.
In another embodiment of the present invention, the chlorine dioxide treatment is performed at a concentration of 8 to 1000 ppm.
In addition, the present invention provides a method for insecticidal action using the above activity increasing method.
The present invention confirms that the acetylcholine ester raising activity is increased by chlorine dioxide treatment, and the insecticidal effect can be obtained by increasing the activity of the acetylcholine ester raise.
This is in contrast to the insecticidal effect of the insecticides of the prior art in confirming the insecticidal effect by inhibiting the activity of AChE. The molecular end point showing the insecticidal effect according to chlorine dioxide treatment is AChE, and the activity of AChE is increased, The composition can be used as an excellent insecticidal composition.
FIG. 1 is a graph showing the results of observing the mortality of a moth moth with different concentrations of chlorine dioxide.
FIG. 2 is a graph showing the results of measurement of mortality by concentration of chlorine dioxide on the seventh day after injection.
FIG. 3 is a graph showing the results of analysis of AChE activity by the developmental stages of the moth.
FIG. 4 is a graph showing the results of confirming the AChE activity according to the concentration of chlorine dioxide in the fifth instar larva of the white moth.
5 is a view showing an experimental apparatus for confirming the change in the negative luminosity of the hanamari moth by chlorine dioxide.
6 is a diagram showing the result of confirming the change in the negative luminosity of the fifth instar larvae of anthocyanin moth by chlorine dioxide.
7 is a graph showing changes in the activity of AChE by fumigation treatment of chlorine dioxide.
8 is a view showing a result of observation of surviving individuals after 5 days of fumigation treatment of chlorine dioxide.
FIG. 9 is a graph showing the results of confirming AChE activity in an individual surviving 5 days after fumigation with chlorine dioxide. FIG.
The present invention relates to a composition for increasing the acetylcholine ester raising activity of insect pests including chlorine dioxide and a method for increasing the acetylcholine ester raising activity for treating the chlorine dioxide.
The concentration of the chlorine dioxide may be 8 to 1000 ppm. The treatment of chlorine dioxide can be effected by intramuscular injection or fumigation, and is preferably carried out at 8 to 800 ppm.
The insect pest may be Plodia interpunctella but is not limited thereto as long as it is an insect showing an increase in the activity of the acetylcholine ester raise by treatment with chlorine dioxide as in the present invention.
The present invention can provide an insecticidal composition comprising the activity-increasing composition, and the composition can be used with the addition of commonly used additives.
In addition, the present invention can provide a method of insecticidal action by increasing the activity of raising acetylcholine ester by treatment with chlorine dioxide.
The control effect of chlorine dioxide on low insect pests is known, but it is expected that this insecticide is ultimately caused by oxidative stress caused by active oxygen production of chlorine dioxide. However, there is no link to what kind of active oxygen generated by chlorine dioxide leads to the insecticidal effect through certain molecular endpoints. Therefore, the present inventors paid attention to AChE as a molecular end point connecting these causal relationships.
On the other hand, in the examples of the present invention, the mortality of the moth moth was confirmed through the liquid phase or the fumigation of chlorine dioxide, and chlorine dioxide increased the activity of AChE in common.
In addition, when AChE increased, it was confirmed that the acetylcholine secretion of the moth moth was controlled, and the normal activity of the nervous system was changed by the regulation of acetylcholine, thereby causing disturbance to the congenital behavior of the moth moth.
Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the examples.
Example One: Disclosure breed
The moth moths used in this example were collected at the dry vegetable storage warehouse of Daegu in 1994 as indoor moths. The larvae were incubated at a temperature of 28 ± 1 ℃, a relative humidity of 65-75%, and a day length of 16 ± 1 ℃ using artificial diet (800 g rice bran, 200 g yeast extract, 500 mL glycerol, 2 g sorbic acid and 2 g methyl p-hydroxybenzoate) : 8 (L: D) h. In order to reduce the random genetic drift of small groups, more than 100 male and female generations were mated to form the next generation. Adults were given 10% sugar water.
Example 2: chlorine dioxide treatment
In this embodiment, 800 ppm of chlorine dioxide was used as a chlorine dioxide (Furugo Farm, Kyonggi Province, Korea).
(1) Liquid phase injection treatment
The chlorine dioxide liquid injection process was performed at 0, 50, 100, 200, 400 and 800 ppm. Injection was performed using a microcapillary (SYS-microcontroller, World Precision Instruments, Sarasota, FL, USA) equipped with a micropump using a glass capillary. Glass capillary tubes were fabricated using micropipette puller (PN-30, Narishige, Tokyo, Japan).
After the liquid phase injection treatment, the cells were allowed to stand at 25 캜 in an indoor condition for 24 hours, and the mortality was examined. Death was defined as the absence of arbitrary behavior when the abdomen was lightly pressed with tweezers.
(2) Fumigation treatment
Chlorine dioxide was produced by electrolysis. The concentration was adjusted because it was dissolved in water and fumed in gas form. It was administered using a gas generator (Purugo Farm, Gyeonggi-do, Korea) in a sealed chamber specially made of acrylic plate. Test insects were administered with a small amount (approximately 10) brown rice in 50 mL tubes (Falcon, Tamaulipas, Mexico). The upper surface of the vessel was covered with mesh to enable fumigation treatment. This insect container was placed in a chlorine dioxide treatment chamber and the concentration of chlorine dioxide was continuously monitored with a gas leak detector (C16, Analytical technology, Collegeville, PA, USA) during the treatment period to confirm the treatment concentration.
The fumigation treatment was carried out for 0, 2, 3, and 4 days, respectively, at a concentration of 50 ppm chlorine dioxide. Each concentration treatment was carried out in three replicates, each of which was performed on 10 5th instar.
After the fumigation, the feces were left to stand for 24 hours at room temperature of 25 ° C. Death was defined as the absence of arbitrary behavior when the abdomen was lightly pressed with tweezers.
Example 3: Toxicity of Chlorine Dioxide by Injection
In the method of Example 2, 1 μL of chlorine dioxide was injected into the fifth instar larva.
3.1. Determination of mortality by concentration
The concentration of chlorine dioxide was adjusted to 0, 50, 100, 200, 400, and 800 ppm, and the mortality rate of the 5th instar larvae was confirmed. The results are shown in FIG.
As shown in Fig. 1, toxicity was confirmed even at the lowest concentration of 50 ppm, which is the analytical concentration. Therefore, the insecticidal power by chlorine dioxide treatment was confirmed.
3.2. Determination of the mortality rate according to the elapsed days
FIG. 2 shows the mortality rate measured up to 7 days after the treatment, and the insecticidal power is also increased as the number of days elapsed after the treatment is increased. After 7 days of treatment, the maximum insecticidal activity was observed. Thereafter, there was no difference in insecticidal activity. Therefore, the toxicity analysis was set to 7 days after the treatment. Half of this specific process, based on the concentration lethal dose (LC 50) was 196.5 ppm (95% CI: 125.6 ~ 301.5) appeared.
Thus, it can be seen that chlorine dioxide injected into the body causes an insecticidal effect on the larvae moth larvae.
Example 4: Growth period according to chlorine dioxide treatment AChE Verify Active
The AChE activity was analyzed by the developmental stage of the moth moth. In Fig. 3, L1-L5 means 1-5 week old larva, P means pupa, and A means adult. From the fully developed first instar larva to adult, all of them showed similar activity except the last order and pupa. During the last period and the pupal period, AChE activity was lower than other developmental stages.
Figure 4 shows the change in AChE activity following the injection of chlorine dioxide to the final 5th instar. At low chlorine dioxide concentrations (0.8 ppm), there was no change in AChE activity, whereas at more than 8 ppm, the activity was increased at least twice, regardless of treatment concentration. Considering that the insecticidal effect (LC 2 ) value of 2% of Hwangryong moth according to chlorine dioxide treatment is 4.86 ppm (95% confidence interval: 0.30-15.54) ppm, 8 ppm treatment of chlorine dioxide reduces physiological disorder The concentration of chlorine dioxide used in the injection of the blood was then set at 8 ppm.
Thus, it was confirmed that AChE activity was increased by treatment with chlorine dioxide. The increase in AChE activity was observed at chlorine dioxide concentrations above 8 ppm, which could lead to such insecticidal effects.
Example 5: Confirming the disturbance of the fluorescence by the chlorine dioxide treatment
In order to examine the behavioral changes of 5th instar moths after chlorine dioxide treatment, we confirmed the disturbance of the negative luminosity. As in the experimental tool shown in Fig. 5, after connecting two 50 mL test tubes, one laboratory tube was covered with a bright environment and the remaining tubes were covered with a black tape to maintain a dark environment. The experiment was conducted by placing 10 larvae in the laboratory.
Chlorine dioxide (8 ppm, injected with blood) was treated and the negative diurnal behavior was analyzed according to the elapsed time. As shown in Fig. 6, the negative diurnal behavior was disturbed as time passed after the treatment. There was no significant difference (P = 0.7139) between the control and the control until 6 hours after the treatment, but there was a large difference (P = 0.0676) between the 12 and 24 hour treatments.
In other words, the increase of AChE activity by chlorine dioxide treatment on H. moth larvae can decrease the amount of acetylcholine which is a neurotransmitter in neuronal synapses, and shows that the moth larvae respond to light and move in the opposite direction, In view of the need for transmission of the body nerve signal through the synaptic connection, the activity of AChE was increased by chlorine dioxide, and it was confirmed that the signal transduction material of neuronal synapse could be easily decomposed and inhibited neurotransmission.
Example 6: According to chlorine dioxide fumigation treatment AChE Check for changes in activity
In this example, changes in the activity of AChE were analyzed for chlorine dioxide fumigation at a concentration of 50 ppm. As shown in Fig. 7, the insecticidal activity was increased as the treatment time of the chlorine dioxide fumigation of 50 ppm was increased with respect to the fifth instar larva of the moth.
The surviving individuals after 5 days of initial treatment were mostly pupa (P) or dedicated phase (PP) (Fig. 8).
Analysis of AChE activity on survivors showed that the chlorine dioxide treatment increased the activity from 2 to 4 times that of the control (Fig. 9).
Through the above example, the fungicidal treatment of chlorine dioxide shows the insecticidal effect of the moth moth, but the molecular end point of one molecule is AChE, and the increase of the activity of the enzyme inhibits physiological behavior disturbance and various body nervous control, leading to death .
It will be understood by those skilled in the art that the foregoing description of the present invention is for illustrative purposes only and that those of ordinary skill in the art can readily understand that various changes and modifications may be made without departing from the spirit or essential characteristics of the present invention. will be. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive.
Claims (9)
Wherein the concentration of chlorine dioxide in the composition is 8 to 1000 ppm.
Wherein the insect pest is Plodia interpunctella. ≪ RTI ID = 0.0 > 11. < / RTI >
Wherein the chlorine dioxide treatment is carried out by intramuscular injection or fumigation.
Wherein the chlorine dioxide treatment is carried out at a concentration of from 8 to 1000 ppm.
Wherein the insect pest is Plodia interpunctella.
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