Bee Antidote Composition

Abstract

A honeybee antidote composition against carbaryl insecticide contains curcumin at a concentration of 30 to 300 ppm.

Description

Technology Field

[0001] The present invention relates to a honeybee antidote composition, and more specifically, to a honeybee antidote composition that provides a supplementary food to increase the survival rate of honeybees poisoned by carbaryl insecticides. Background Technology

[0003] Honeybees are important pollinating insects. Damage to honeybees caused by pesticide poisoning is very serious. In particular, pesticide poisoning of honeybees in the spring in apple orchards is mostly caused by the insecticide carbaryl.

[0004] Carbaryl, a synthetic carbamate acetylcholinesterase inhibitor suspected of being an endocrine disruptor, is commonly used in agriculture for pest control due to its broad spectrum and low toxicity to mammals. Its versatility extends to lawns and garden environments. However, deforestation and potential damage, particularly to bees, highlight environmental issues. While effective, the use of carbaryl underscores the need for increased scrutiny of its impact on ecosystems and the exploration of responsible pest management practices and alternative solutions.

[0005] Like all living organisms, honeybees possess detoxification mechanisms to neutralize and eliminate harmful substances. Studies have shown that enzyme groups such as P450, GST, and COE are involved in the detoxification of pesticides and secondary metabolites in honeybees. Honeybees activate these detoxification mechanisms when exposed to toxic pesticides.

[0006] However, there is a problem in that the continuous exposure of honeybees to various pesticides and toxic substances remains a major cause of the decline in the honeybee population.

[0007] delete Prior art literature

[65535] Korean Patent Publication No. 10-1079250 Biomarker for confirming carbaryl exposure and method for confirmation using the same (Registration Date: Oct. 27, 2011) The problem to be solved

[0008] Therefore, the objective of the present invention is to provide a honeybee antidote composition capable of serving as an effective antidote for pesticide toxicity in honeybees. means of solving the problem

[0010] The honeybee antidote composition for carbaryl insecticide according to the present invention for achieving the above objective comprises a curcumin concentration of 30 to 300 ppm. It is possible to provide a curcumin concentration of 30 to 300 ppm that can increase the survival rate of honeybees poisoned by carbaryl insecticide and serve as an effective antidote for insecticide toxicity in honeybees.

[0011] Here, having a curcumin concentration of 90 to 110 ppm is desirable because it can increase the survival rate of honeybees poisoned by the carbaryl insecticide and provide the most desirable curcumin concentration of 90 to 110 ppm, which can act as an effective antidote for insecticide toxicity in honeybees.

[0012] Meanwhile, a method for reducing toxic damage from carbaryl insecticide to honeybees comprises the steps of: preparing a 50% aqueous sucrose solution; including curcumin at a concentration of 30 to 300 ppm in the prepared 50% aqueous sucrose solution; and providing the 50% aqueous sucrose solution containing curcumin as honeybee feed, thereby increasing the survival rate of honeybees poisoned by carbaryl insecticide and providing a concentration of curcumin at 30 to 300 ppm that can act as an effective antidote for insecticide toxicity in honeybees.

[0013] And the step of including the curcumin at a concentration of 30 to 300 ppm is desirable because if the curcumin is included in the 50% sucrose aqueous solution at a concentration of 90 to 110 ppm, it can increase the survival rate of honeybees poisoned by carbaryl and provide the most desirable concentration of curcumin at 90 to 110 ppm, which can act as an effective antidote for pesticide toxicity in honeybees. Effects of the invention

[0015] According to the present invention, the most desirable concentration of curcumin is provided at 90 to 110 ppm, which increases the survival rate of honeybees poisoned by the carbaryl insecticide and has the effect of acting as an effective antidote for insecticide toxicity in honeybees.

[0016] In addition, it can increase the survival rate of honeybees poisoned by the insecticide carbaryl, enhance the detoxification role of honeybees against insecticide toxicity, and have the effect of rapidly increasing the activity of oxidases that perform detoxification. Brief explanation of the drawing

[0018] Figure 1 is a flowchart of a method for reducing toxic damage from carbaryl insecticides to honeybees according to the present invention. Figure 2 is a flowchart of a method to reduce toxic damage from carbaryl insecticides to other honeybees. Figure 3 is a graph showing the effect of applying various concentrations of curcumin on the survival of locally carbaryl-poisoned honeybees (A) and collectors (B). Figure 4 is a graph of the Kaplan-Meier survival curves of experimental honeybees (Apis mellifera) chronically orally exposed to 2.5 ppm and 5 ppm of carbaryl. Figure 5 is a graph showing the mortality rate of honeybees over 10 days after chronic exposure to 2.5 ppm (A) and 5 ppm (B) of carbaryl. Figure 6 is a graph showing the effect of applying a 100 ppm curcumin supplement on the honeycomb area (A-B) and number of workers (C-D) of beehives that received a curcumin (100 ppm) supplement solution (A, C) and a regular sugar solution (B, D). Figure 7 is a graph showing the effect of applying a 100 ppm curcumin supplement on the honey area (A-B) and colony weight (C-D) of urticaria treated with curcumin supplement solution (A, C) and regular sugar solution (B, D). Figures 8 and 9 are graphs showing the gene expression analysis of detoxification enzymes 24 hours after exposure to carbaryl for SU (50% sugar syrup), CU (50% sugar syrup + 200 ppm curcumin), and the control group (ACET, exposed to only 2 µl of acetone and supplied with sugar). Specific details for implementing the invention

[0019] Hereinafter, a honeybee antidote composition according to a preferred embodiment of the present invention will be described in detail with reference to the attached drawings.

[0020] FIG. 1 is a flowchart of a method for reducing toxic damage from carbaryl insecticides to honeybees according to the present invention, FIG. 2 is a flowchart of another method for reducing toxic damage from carbaryl insecticides to honeybees, FIG. 3 is a graph showing the effect of applying curcumin at various concentrations on the survival of a honeybee (A) and a collector (B) locally poisoned with carbaryl, FIG. 4 is a graph of the Kaplan-Meier survival curve of experimental honeybees (Apis mellifera) chronically orally exposed to 2.5 ppm and 5 ppm of carbaryl, FIG. 5 is a graph showing the honeybee mortality rate over 10 days after chronic exposure to 2.5 ppm (A) and 5 ppm (B) of carbaryl, FIG. 6 is a graph showing the honeycomb area (A-B) of beehives receiving curcumin (100 ppm) supplement solutions (A, C) and ordinary sugar solutions (B, D) and the worker after the application of a 100 ppm curcumin supplement. Figure 7 is a graph showing the effect on the number (C-D), Figure 7 is a graph showing the effect of applying a 100 ppm curcumin supplement on the honey area (A-B) and colony weight (C-D) of urticaria treated with curcumin supplement solution (A, C) and regular sugar solution (B, D), and Figures 8 and 9 are graphs showing the gene expression analysis of detoxification enzymes 24 hours after carbaryl exposure for SU (50% sugar syrup), CU (50% sugar syrup + 200 ppm curcumin), and the control group (ACET, exposed to only 2 µl of acetone and supplied with sugar).

[0021] Referring to FIGS. 1 to 9, a honeybee antidote composition and a method for reducing toxic damage from carbaryl insecticides to honeybees will be explained.

[0022] A honeybee antidote composition for carbaryl insecticides contains a concentration of 30 to 300 ppm of curcumin, which can increase the survival rate of honeybees poisoned by carbaryl insecticides and act as an effective antidote for insecticide toxicity in honeybees.

[0023] A honeybee antidote composition for carbaryl insecticides may include the most desirable concentration of curcumin at 90 to 110 ppm, which can increase the survival rate of honeybees poisoned by carbaryl insecticides and serve as an effective antidote for insecticide toxicity in honeybees. Here, curcumin can rapidly increase the activity of oxidases that perform the detoxification role.

[0024] This invention describes an experimental method that can increase the survival rate of honeybees poisoned by the carbamate insecticide Carbaryl and serve as an effective antidote for insecticide toxicity in honeybees.

[0025] Curcumin is selected after screening various substances capable of detoxifying pesticide toxicity.

[0026] After exposing adult honeybees to pesticides through feeding and contact, we investigate whether feeding a candidate antidote has a mortality reduction effect.

[0027] Subsequently, through concentration treatment, the concentration at which curcumin selectively detoxifies carbaryl toxicity is investigated.

[0028] Long-term laxity and toxicity reduction capabilities are evaluated through long-term exposure studies.

[0029] Evaluate whether the application of 100 ppm curcumin to bee colony health is harmful or detoxifying under greenhouse conditions.

[0030] Afterwards, additional experiments on the activity of detoxification enzymes are conducted to investigate whether the activity of detoxification enzymes is high at 100 ppm of curcumin.

[0031] Figure 1 is a flowchart of a method for reducing toxic damage from carbaryl insecticides to honeybees according to the present invention.

[0032] Prepare a 50% sucrose solution (S1).

[0033] Curcumin is added to a prepared 50% aqueous sucrose solution at a concentration of 30 to 300 ppm (S2).

[0034] A 50% sucrose solution containing curcumin is provided as bee food (S3).

[0035] Figure 2 is a flowchart of a method to reduce toxic damage from carbaryl insecticides to other honeybees.

[0036] Prepare a 50% sucrose solution (S11).

[0037] Curcumin at a concentration of 90 to 110 ppm is added to a prepared 50% aqueous sucrose solution (S12).

[0038] A 50% sucrose solution containing curcumin is provided as bee food (S13).

[0040] To evaluate the effects on the survival of honeybees intentionally poisoned with carbaryl, various concentrations of curcumin (50, 100, and 200 ppm) were administered. This experiment was conducted separately on emerging honeybees and hunter-gatherers. 0.063 µg of the insecticide was administered to the thorax of emerging honeybees, while 0.11 µg was administered per bee to hunter-gatherers. After exposure to carbaryl, supplemental food was provided to the intoxicated honeybees. The control group consumed only sugar syrup, whereas the experimental group consumed supplemental food containing curcumin. All honeybee groups were fed for 48 hours under controlled laboratory conditions.

[0041] The effects of adding 100 ppm of curcumin to melon production greenhouses were evaluated. Thirty greenhouses owned by three farmers were used in this study. One group of bees was provided with a supplemental solution of 100 ppm curcumin (CU100), while control hives were provided with only 50% sugar syrup. Colony strength factors were measured every 10 days.

[0042] As a result, it was confirmed that there was a reduction in mortality rates in adult honeybees (brood bees, foraging bees) after they were exposed to pesticides through contact and fed with the antidote candidate.

[0043] The same results were confirmed through long-term drug contact and the administration of an antidote.

[0044] The results of the field experiment also showed that honeybee activity was superior in the curcumin 100 ppm treatment group.

[0045] Therefore, it has been proven that a cucumin concentration of 100 ppm has a selective detoxification effect on carbaryl toxicity.

[0046] Subsequently, additional experiments on the activity of detoxification enzymes confirmed that the activity of detoxification enzymes was high even at 200 ppm of cucumin.

[0047] In laboratory tests, the control group administered only sugar syrup showed a high mortality rate of 78% among bees exposed to carbaryl. A noticeable decrease in the mortality rate of nurse bees was observed after administering various concentrations of curcumin. The significance of this reduction became even more pronounced when higher doses of the antidote, up to 100 ppm, were applied to newly emerged bees. In parallel feeding experiments conducted on hunter-gatherers, individuals treated with CU50 and CU100 showed significantly lower mortality rates compared to the control group. Our analysis indicates that applying CU100 after feeding can serve as an effective antidote to mitigate the adverse effects of carbaryl on intoxicated bees.

[0048] In field tests, the effects of CU100 on colony strength factors of Seongju melon pollination colonies were evaluated. The results showed that the breeding area tended to increase in the group receiving CU100, whereas it tended to decrease in the control group. Similarly, the number of workers showed a slight increasing trend in both groups. The honey area remained consistent throughout the study, increasing slightly in hives supplied with CU100 and decreasing slightly in hives supplied with only sugar syrup. Despite an overall decrease in colony weight in both groups, the colonies supplied with CU100 experienced a less steep decline compared to the colonies supplied with only sugar syrup.

[0049] In conclusion, the study included the application of curcumin to mitigate the negative effects of pesticides on honeybees. Curcumin was selected due to its well-known biological activities, including anti-inflammatory, antioxidant, and detoxification properties. Our goal was to evaluate its effect on reducing honeybee mortality following pesticide exposure.

[0050] Experimental results on honeybees exposed to carbaryl (0.063 µg / bee) and nurse honeybees subsequently fed 100 ppm of curcumin showed a remarkable reduction in mortality. While the mortality rate in the control group was 78%, it decreased to 28% in the CU100 group. In the case of hunter-gatherers, the control group showed a mortality rate of 34%, which dropped significantly to 18% and 16% in the groups that received CU50 and CU100 after feeding.

[0051] Field trials of CU100 on bee colonies used for melon pollination showed a positive effect on colony strength factors compared to colonies that received only sugar syrup. Our findings suggest that curcumin can serve as an effective antidote for pesticide toxicity in bees. Adding this compound to food can significantly increase survival chances in environments exposed to pesticides. Apple pollination service providers are advised to include Cu in the sugar solutions provided to bees. Considering the positive effects of curcumin on bee colonies involved in melon greenhouse pollination, it is recommended to include Cu as a dietary supplement.

[0053] Figure 3 is a graph showing the effect of applying various concentrations of curcumin on the survival of locally carbaryl-poisoned honeybees (A) and collectors (B).

[0054] In an indoor feeding study on brood bees and worker bees, 100 ppm of curcumin stably reduced the mortality rate by 80 24, 35 It was lowered to 18. In indoor feeding tests on brood bees and foraging bees, curcumin 50 ppm also lowered the mortality rate to 80 50, 35 It was lowered to 24. However, it did not show the same effect as curcumin 100 ppm.

[0055] Here, looking at the mortality rates at curcumin 90 ppm, curcumin 100 ppm, and curcumin 110 ppm in indoor feeding tests on brood bees and worker bees, curcumin 90 ppm showed a better effect than curcumin 50 ppm (80 33, 35 24) indicates. However, the mortality rate at curcumin 110 ppm shows a phenomenon of increasing again (80 34, 35 24) was shown.

[0056] Figure 4 is a graph of the Kaplan-Meier survival curves of experimental honeybees (Apis mellifera) chronically orally exposed to 2.5 ppm and 5 ppm of carbaryl.

[0057] The Su group received only 50% sugar syrup, and the Cu group received only 50% sugar syrup + 100 ppm curcumin.

[0058] In a long-term indoor feeding experiment, treatment with Cu (Curcumiin 100 ppm) did not show toxicity, and in the carbaryl exposure test, the survival rate of honeybees was highest with treatment with curcumin 100 ppm.

[0059] Figure 5 is a graph showing the mortality rate of honeybees over 10 days after chronic exposure to 2.5 ppm (A) and 5 ppm (B) of carbaryl.

[0060] The control group was administered only 50% sugar syrup, and the Cu group was administered 50% sugar syrup + 100 ppm curcumin.

[0061] In an indoor long-term exposure test of carbaryl, treatment with 100 ppm curcumin reduced honeybee mortality by 38 21, 78 It showed an effect of reducing it by about 55%.

[0062] Figure 6 is a graph showing the effect of applying a 100 ppm curcumin supplement on the honeycomb area (A-B) and number of workers (C-D) of beehives that received a curcumin (100 ppm) supplement solution (A, C) and a regular sugar solution (B, D).

[0063] In a long-term exposure test in a Seongju Korean melon greenhouse, both adult and larval honeybees showed an increasing trend in the group treated with 100 ppm curcumin, whereas larvae decreased in the colony treated only with sucrose solution.

[0064] Figure 7 is a graph showing the effect of applying a 100 ppm curcumin supplement on the honey area (A-B) and colony weight (C-D) of urticaria treated with curcumin supplement solution (A, C) and regular sugar solution (B, D).

[0065] In the long-term exposure test in the Seongju Korean melon greenhouse, there was no change in honey weight, but the rate of hive weight reduction was lower in the curcumin 100 ppm treatment group than in the colony treated only with sucrose solution.

[0066] Figures 8 and 9 are graphs showing the gene expression analysis of detoxification enzymes 24 hours after exposure to carbaryl for SU (50% sugar syrup), CU (50% sugar syrup + 200 ppm curcumin), and the control group (ACET, exposed to only 2 µl of acetone and supplied with sugar).

[0067] Figure 8 shows that the activity of oxidase, which plays a detoxification role in the intestines of honeybees, increased rapidly in the Cu-treated group compared to the untreated group.

[0068] As can be seen in Figure 11, the activity in the heads of honeybees was similar to that of the untreated group, and the activity was very low in the area where sucrose solution was added after carbaryl treatment.

[0069] Due to the above-described honeybee antidote composition and the method for reducing toxic damage from carbaryl insecticides to honeybees, the most desirable concentration of curcumin is provided at 90 to 110 ppm, which increases the survival rate of honeybees poisoned by carbaryl insecticides and can serve as an effective antidote for insecticide toxicity in honeybees.

[0070] In addition, it can increase the survival rate of honeybees poisoned by the carbaryl insecticide, enhance the detoxification role of honeybees against insecticide toxicity, and rapidly increase the activity of oxidases that perform detoxification.

Claims

Claim 1 A honeybee antidote composition for carbaryl insecticides, wherein curcumin is selected after screening various substances capable of detoxifying pesticide toxicity, adult honeybees are exposed to pesticides through feeding and contact, and then a mortality reduction effect is investigated through feeding of the antidote candidate, subsequently the concentration at which curcumin selectively detoxifies carbaryl toxicity is investigated through concentration treatment, long-term phytotoxicity and toxicity reduction ability are evaluated through long-term exposure studies, and harmfulness to colony health and detoxification are evaluated by applying curcumin at a concentration of 90 to 110 ppm under greenhouse conditions, and subsequently additional experiments on detoxification enzymes are conducted to investigate whether the activity of detoxification enzymes is high at a curcumin concentration of 90 to 110 ppm, and the honeybee antidote composition is characterized by containing curcumin at a concentration of 90 to 110 ppm. Claim 2 delete Claim 3 A method for mitigating damage from carbaryl toxicity in honeybees comprises the following steps: screening various substances capable of detoxifying pesticide toxicity and selecting curcumin; investigating whether there is a mortality reduction effect by exposing adult honeybees to pesticides via contact and having them consume the antidote candidate; subsequently investigating the concentration at which curcumin selectively detoxifies carbaryl toxicity through concentration treatment; evaluating long-term phytotoxicity and toxicity reduction capabilities through long-term exposure studies; evaluating whether it is harmful to colony health or detoxifies by applying curcumin at a concentration of 90 to 110 ppm under greenhouse conditions; subsequently conducting additional experiments on detoxification enzymes to investigate whether the activity of detoxification enzymes is high at a curcumin concentration of 90 to 110 ppm; and preparing a 50% sucrose aqueous solution. A method for reducing toxic damage from carbaryl insecticides to honeybees, characterized by including the step of including curcumin at a concentration of 90 to 110 ppm in the above 50% sucrose aqueous solution. Claim 4 delete

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