Mosquito prevention and control composition based on jonquil extract and application of mosquito prevention and control composition

Abstract

The invention discloses a mosquito prevention and control composition based on jonquil extract and application of the mosquito prevention and control composition. The invention provides an application of an ethanol extract of jonquil leaves in killing mosquitoes or preparing products for killing mosquitoes. The invention firstly proves that the kalanchoe blossfeldiana ethanol extract has a killing effect on three mosquito species, and the killing effect is obviously superior to that of kalanchoe laciniata which is the same family plant. The invention further provides a mosquito prevention and control composition containing the ethanol extract of the jonquil leaves and a mosquito prevention and control compound composition containing the ethanol extract of the jonquil leaves and dinotefuran. The insecticide poison bait prepared by compounding the jonquil extract and the chemical insecticide provides a new thought for constructing an efficient, low-consumption and environment-friendly mosquito comprehensive treatment system, and is suitable for comprehensive prevention and control of mosquito-borne diseases.

Description

Technical Field

[0001] This invention belongs to the field of biological pesticide technology, specifically relating to a mosquito control composition based on an extract of Kalanchoe blossfeldiana, a plant of the Crassulaceae family, and its application. Background Technology

[0002] Mosquitoes can transmit a variety of viral diseases that pose a serious threat to public health, including dengue fever, yellow fever, Japanese encephalitis, West Nile fever, Zika virus disease, chikungunya, and Sindbis disease. While traditional chemically synthesized insecticides (such as organophosphate pesticides) are effective in controlling mosquito populations, their long-term use has led to increasing resistance, ecotoxicity, and residual pollution. Against this backdrop, plant-derived pesticides have become a research hotspot in mosquito vector control due to their environmental friendliness, target specificity, and low likelihood of inducing resistance.

[0003] In recent years, with the deepening research on the active ingredients of traditional Chinese medicine, more and more plant-derived pesticides have been discovered and applied to mosquito control. For example, artemisinin, an active ingredient extracted from Artemisia annua, not only has antimalarial effects but has also been found to have a significant toxic effect on mosquito larvae. Osthol from Cnidium monnieri has broad biological activity and has long been used as a pesticide in my country, showing insecticidal activity against the larvae of various mosquitoes, including Aedes aegypti and Culex pipiens pallens. Although plant-derived pesticides have shown great potential in mosquito control, their practical application still faces many challenges, such as the purification cost of active ingredients, chemical stability, and the need to comprehensively consider the mechanism of action. Finding new, highly effective, low-toxicity, and environmentally friendly plant-derived pesticides remains an important research direction. Kalanchoe (Kalanchoe blossfeldiana) Kalanchoe blossfeldiana Kalanchoe blossfeldiana is a perennial succulent plant belonging to the genus Kalanchoe in the family Crassulaceae. Native to Madagascar, it is now widely cultivated as an ornamental plant in temperate and subtropical regions worldwide. Its cymose inflorescences, which bloom in winter and spring, are richly colored, including red, pink, yellow, and white, giving it high horticultural value. While Crassulaceae plants are known to contain antibacterial active ingredients (such as flavonoids), their potential for mosquito control has not yet been fully explored.

[0004] Attractive Toxic Sugar Baits (ATSB) technology is an innovative mosquito control strategy. This technology utilizes the mosquito's (both male and female) need to ingest sugar sources for energy. Highly effective attractants (such as nectar or fruit juice) are mixed with low-toxicity insecticides to create bait. After being attracted to the attractant and feeding on it, mosquitoes die from insecticide poisoning. ATSB technology has advantages such as strong targeting, broad range of action, and environmental friendliness. Developing plant-based ATSB systems by combining plant-derived insecticides with ATSB technology is a research hotspot for achieving green and sustainable mosquito control and has broad application prospects. Summary of the Invention

[0005] The purpose of this invention is to provide a mosquito control bait based on Kalanchoe blossfeldiana extract and its application. Experiments have shown that the Kalanchoe blossfeldiana ethanol extract alone has significant killing activity against Culex pipiens quinquefasciatus, Aedes albopictus, and Anopheles sinensis, with mortality rates of 55.56%, 55.00%, and 63.33% respectively after 48 hours. Through further compounding, a novel compounding system of Kalanchoe blossfeldiana plant extract and chemical insecticide (dinotefuran) was developed: 0.1% Kalanchoe blossfeldiana extract + 10... -3 The g / L fipronil compound group increased the 48-hour mortality rate of Culex pipiens quinquefolius to 89.50% in simulated field tests. The resulting highly efficient Kalanchoe blossfeldiana extract combined with chemical insecticides provides a new approach for building an efficient, low-cost, and environmentally friendly integrated mosquito control system, suitable for integrated prevention and control of mosquito-borne diseases.

[0006] To achieve the above objectives, the present invention adopts the following technical solution: In a first aspect, the present invention provides the use of an ethanol extract of Kalanchoe blossfeldiana leaves in the killing of mosquitoes or in the preparation of products for killing mosquitoes.

[0007] The inventors further compared the effects of oral administration of Kalanchoe blossfeldiana ethanol extract, Sedum spectabile ethanol extract, and Kalanchoe lanceolata ethanol extract on mosquito mortality. The results showed that Kalanchoe blossfeldiana ethanol extract exhibited stable and significant killing activity against multiple major mosquito vectors, demonstrating the best mosquito-killing activity. This invention is the first to confirm the killing effect of Kalanchoe blossfeldiana ethanol extract on three major mosquito species (Culex pipiens quinquefasciatus, Aedes albopictus, and Anopheles sinensis).

[0008] According to an embodiment of the present invention, the Kalanchoe blossfeldiana ethanol extract is obtained through the following steps: Kalanchoe blossfeldiana leaves are crushed and then soaked in ethanol, followed by separation and collection of the extract. As an example, the crushing is performed by chopping. As an example, the soaking is performed overnight at room temperature. As an example, the amount of ethanol used is sufficient to submerge the leaves; for example, if 10 kg of Kalanchoe blossfeldiana leaves are used, the liquid level should be approximately 3 cm above the leaves. As an example, the separation and collection of the extract includes rotary evaporating the soaking solution at 50°C until no ethanol residue remains.

[0009] According to embodiments of the present invention, the mosquito is selected from one or more of Culex pipiens quinquefolius, Aedes albopictus, and Anopheles sinensis. The mosquito is either male or female.

[0010] According to embodiments of the present invention, the killing is achieved through ingestion poisoning and / or contact killing. The inventors further investigated the effect of direct contact with the ethanol extract of Kalanchoe blossfeldiana on mosquito mortality, finding that the extract possesses both enterotoxic and contact-killing effects.

[0011] In one embodiment of the present invention, the killing is a feeding poisoning method, in which mosquitoes are fed with an ethanol extract of Kalanchoe blossfeldiana.

[0012] In another embodiment of the invention, the killing is a contact killing, which involves bringing mosquitoes into contact with the Kalanchoe ethanol extract.

[0013] Secondly, the present invention provides a mosquito control composition comprising a sugar source, an ethanol extract of Kalanchoe blossfeldiana leaves, and a solvent.

[0014] According to an embodiment of the present invention, the sugar source is selected from one or more of sucrose, honey, fructose, and glucose; it is understood that commercially available sugar-providing products can be used as sugar sources. According to an embodiment of the present invention, the concentration of the sugar source in the composition is 5% to 15% by mass-volume percentage (g:mL), such as 8% (i.e., 8g sucrose: 100mL ethanol).

[0015] According to an embodiment of the present invention, the concentration of the ethanol extract of Kalanchoe blossfeldiana leaves in the composition is 0.1% to 1%, such as 1% (i.e., 1g Kalanchoe blossfeldiana extract: 100mL ethanol), by mass-volume percentage (g:mL).

[0016] According to an embodiment of the present invention, the solvent may be ethanol.

[0017] It is understood that the mosquito control composition may also include food coloring and / or preservatives.

[0018] Thirdly, the present invention provides a mosquito control compound composition comprising a sugar source, an ethanol extract of Kalanchoe blossfeldiana leaves, fipronil, and fruit juice.

[0019] According to an embodiment of the present invention, the sugar source is selected from one or more of sucrose, honey, fructose, and glucose; it is understood that commercially available sugar-providing products can be used as sugar sources.

[0020] According to an embodiment of the present invention, the concentration of the sugar source in the composition is 5% to 15% by mass-volume percentage (g:mL), such as 8% (i.e., 8g sucrose: 100mL fruit juice).

[0021] According to an embodiment of the present invention, the concentration of the ethanol extract of Kalanchoe blossfeldiana leaves in the compound composition is 0.1% to 1%, such as 1% (i.e., 1g Kalanchoe blossfeldiana extract: 100mL fruit juice), by mass-volume percentage (g:mL).

[0022] According to an embodiment of the present invention, the concentration of fipronil in the compound composition is 1×10⁻⁶. -4 g / L~1×10 -2 g / L, including but not limited to 5×10 -4 g / L.

[0023] According to an embodiment of the present invention, the mass ratio of the Kalanchoe ethanol extract to fipronil is (2000-10):1, such as 2000:1.

[0024] According to an embodiment of the present invention, the juice is apple juice or pear juice.

[0025] According to embodiments of the present invention, the compound composition further includes food coloring and / or preservatives. Further, the volume percentage of the food coloring in the compound composition is 0.01% to 0.5%, such as 0.1%; the mass-volume percentage of the preservative in the compound composition is 0.01% to 0.5% (g / mL), such as 0.1%; further, the preservative may be potassium sorbate.

[0026] Fourthly, the present invention provides the application of an ethyl acetate extract of the ethanol extract of Kalanchoe blossfeldiana leaves in the killing of mosquitoes or in the preparation of products for killing mosquitoes; wherein the preparation steps of the ethyl acetate extract of the ethanol extract of Kalanchoe blossfeldiana leaves include: 1) adding water and petroleum ether to the ethanol extract of Kalanchoe blossfeldiana leaves, mixing thoroughly, allowing it to stand and separate into layers, and collecting the lower aqueous phase; 2) adding ethyl acetate to the aqueous phase for extraction, mixing thoroughly, allowing it to stand and separate into layers, collecting the upper ethyl acetate phase, evaporating to dryness, and obtaining the ethyl acetate extract.

[0027] Through screening, the inventors discovered that the ethyl acetate extract of Kalanchoe blossfeldiana resulted in a significantly higher mortality rate in Culex pipiens quinquefolius than other polar solvent extracts.

[0028] In an embodiment of the present invention, in step 1), a solvent is added in a volume ratio of extract:water:petroleum ether of 1:1:3, and in step 2), the volume of ethyl acetate is 3 times that of the aqueous phase.

[0029] Fifthly, the present invention provides a mosquito control composition comprising an ethyl acetate extract of an ethanol extract of Kalanchoe blossfeldiana leaves and a solvent; wherein the preparation steps of the ethyl acetate extract of the ethanol extract of Kalanchoe blossfeldiana leaves include: 1) adding water and petroleum ether to the ethanol extract of Kalanchoe blossfeldiana leaves, mixing thoroughly, allowing the mixture to stand and separate into layers, and collecting the lower aqueous phase; 2) adding ethyl acetate to the aqueous phase for extraction, mixing thoroughly, allowing the mixture to stand and separate into layers, collecting the upper ethyl acetate phase, evaporating to dryness, and obtaining the ethyl acetate extract.

[0030] According to an embodiment of the present invention, the concentration of the ethanol extract of Kalanchoe blossfeldiana leaves in the composition is 0.1% to 1%, such as 1% (i.e., 1g Kalanchoe blossfeldiana extract: 100mL ethanol), by mass-volume percentage (g:mL).

[0031] According to an embodiment of the present invention, the solvent may be ethanol.

[0032] According to embodiments of the present invention, the composition may further include food coloring and / or preservatives.

[0033] In a sixth aspect, the present invention provides a mosquito control method, comprising the following steps: deploying the mosquito control composition (containing an ethanol extract of Kalanchoe blossfeldiana leaves), or the mosquito control compound composition (containing an ethyl acetate extract of the ethanol extract of Kalanchoe blossfeldiana leaves), as described in any of the above claims, in mosquito breeding grounds or habitats to attract and kill mosquitoes.

[0034] According to embodiments of the present invention, the mosquito is selected from one or more of Culex pipiens quinquefolius, Aedes albopictus, and Anopheles sinensis. The mosquito is either male or female.

[0035] According to embodiments of the present invention, in addition to the active ingredients described above, the mosquito control composition or the mosquito control compound composition may also include excipients, which may be selected according to the dosage form. As an example, to facilitate mosquito biting and ingestion, the liquid composition uses cotton balls or sponges as carriers.

[0036] In an embodiment of the present invention, a simulated field experiment was conducted using a mosquito net.

[0037] The present invention has the following beneficial effects: 1. Innovative active ingredients: For the first time, the ethanol extract of Kalanchoe blossfeldiana has been shown to kill three major mosquito species (Culex pipiens quinquefolius / Aedes albopictus / Anopheles sinensis) with a 48-hour mortality rate of 55%-63.33%, which is significantly better than Kalanchoe blossfeldiana (4.44%) and Sedum spectabile (7.78%).

[0038] 2. Synergistic Compound Technology: A binary compound system of "plant-chemistry" is created, consisting of 0.1% Kalanchoe extract + 10... -3 The 48-hour mortality rate of the 100-g / L dinotefuran compound group against female Culex pipiens quinquefolius reached 86.33%, which was 39% higher than that of the dinotefuran single group (p<0.05).

[0039] 3. Optimization of extraction process: A n-butanol fractional extraction method was developed, with an active fraction yield of 0.16% (only 0.05% for ethyl acetate fraction), which solved the problem of enrichment of hydrophilic components.

[0040] 4. Mechanism of action: The ethyl acetate extract of Kalanchoe blossfeldiana showed a significantly higher killing effect on Culex pipiens quinquefolius (90%) than that of petroleum ether extract (16.67%) and n-butanol extract (55.56%), and liquid chromatography-mass spectrometry analysis suggested that it was related to alkaloid regulation. Attached Figure Description

[0041] Figure 1This invention demonstrates the killing effect of different plant extracts on Culex pipiens quinquefolius and Aedes albopictus in Example 1.

[0042] Figure 2 This invention demonstrates the direct contact killing effect of different concentrations of Kalanchoe blossfeldiana extract on Culex pipiens quinquefolius and Aedes albopictus in Example 2 of this invention.

[0043] Figure 3 This demonstrates the killing effect of the combination of Kalanchoe blossfeldiana and insecticide in Example 3 of the present invention on Culex quinquefolia, Aedes albopictus, and Anopheles sinensis.

[0044] Figure 4 This demonstrates the killing effect of the combination of Kalanchoe blossfeldiana and insecticide in the mosquito net in Example 4 of this invention on Culex pipiens quinquefolia.

[0045] Figure 5 This figure shows the killing effect of Kalanchoe blossfeldiana extracts with different polarity solvents on Culex pipiens quinquefolius in Example 5 of the present invention. Note: Figures (A-C) show the mortality rates of male, female, and total Culex pipiens quinquefolius mosquitoes after 24 h and 48 h after oral administration of Kalanchoe blossfeldiana extracts with different polarity solvents. *P<0.05.

[0046] Note: Figure 1-5 In this study, to examine differences in mortality rates among multiple groups, if the data conformed to a normal distribution and had homogeneous variances, one-way ANOVA was used. If the data conformed to a normal distribution but had homogeneous variances, Brown-Forsythe and Welch's ANOVA were used for comparison. If the data did not conform to a normal distribution, the nonparametric Kruskal-Wallis test and Dunn's multiple comparison analysis were used. Detailed Implementation

[0047] The present invention will now be described in further detail with reference to specific embodiments. The given embodiments are merely illustrative of the invention and not intended to limit its scope. The embodiments provided below can serve as a guide for further improvements by those skilled in the art and do not constitute a limitation on the invention in any way.

[0048] Unless otherwise specified, the methods used in the following embodiments are conventional methods, performed according to the techniques or conditions described in the literature in this field or according to the product instructions. Unless otherwise specified, the materials and reagents used in the following embodiments are commercially available.

[0049] The three plants used in the following examples—Kalanchoe blossfeldiana, Sedum spectabile, and Kalanchoe sinensis—were all purchased from Xinzhichun Landscape Gardening Co., Ltd., Suqian City, Jiangsu Province. The sensitive strains of the experimental insects—Culex quinquefasciatus, Aedes albopictus, and Anopheles sinensis—were provided by the Vector Breeding Laboratory of the Academy of Military Medical Sciences. Apple juice was purchased from Beijing Huiyuan Beverage & Food Group Co., Ltd., adhering to enterprise standard QMAAD 0001S Fruit and Vegetable Juices and Fruit and Vegetable Juice Beverages. Pear juice and other fruit juices were used in the same way as apple juice. All fruit juices used were pure fruit juices, without added sugars or preservatives.

[0050] In the following examples, all experimental data were statistically analyzed using SPSS or GraphPad Prism software. Abbott's formula was used to correct for mortality rates: Corrected mortality rate (%) = [(Treatment group mortality rate - Control group mortality rate) / (100 - Control group mortality rate)] × 100. Chi-square test or Fisher's exact test was used to compare differences in mortality rates between different treatment groups. p < 0.05 was considered statistically significant.

[0051] Preparation Example 1: Preparation of Kalanchoe blossfeldiana ethanol extract and extracts from different parts This embodiment provides the preparation steps of Kalanchoe blossfeldiana extract, as follows: Take 10 kg of Kalanchoe blossfeldiana leaves, chop them thoroughly using a herb chopper, then add 95% ethanol to submerge the leaves, ensuring the liquid level is about 3 cm above the leaves. Soak overnight at room temperature. The next day, rotary evaporate the soaking solution at 50°C until no ethanol residue remains, obtaining a crude extract (yield 0.23%). Take 1 / 10 of the extract by volume as the ethanol extract fraction and retain it, then rotary dry for storage.

[0052] The remaining extracts were then subjected to solvent extraction for separation: First, solvent was added at a volume ratio of extract:water:petroleum ether of 1:1:3. After thorough mixing, the mixture was allowed to stand and separate into layers. The upper petroleum ether phase was collected and evaporated to dryness at 50°C to obtain the petroleum ether fraction (v / v) extraction. The petroleum ether fraction was collected (yield 0.08%).

[0053] Next, the lower aqueous phase was extracted with 3 times its volume of ethyl acetate. After thorough mixing, the mixture was allowed to stand and separate into layers. The upper ethyl acetate phase was collected and evaporated to dryness by rotary evaporation to obtain the ethyl acetate fraction (yield 0.05%). The remaining lower aqueous phase was then extracted with 3 times its volume of n-butanol. After thorough mixing, the mixture was allowed to stand and separate into layers. The upper n-butanol phase was collected and evaporated to dryness by rotary evaporation at 50°C to obtain the n-butanol fraction (yield 0.16%). Finally, the remaining lower aqueous phase is evaporated to dryness at 60°C to obtain the water fraction.

[0054] Solid products were obtained from the ethyl acetate and n-butanol fractions. The remaining fractions (ethanol and petroleum ether fractions) were weighed and then ultrasonically dispersed and suspended in ethanol to obtain corresponding 1 g / mL extract solutions. All samples were properly preserved for subsequent analysis of active ingredients and evaluation of bioactivity.

[0055] Comparative Example 1: Preparation of Rhodiola rosea extract and extracts from different parts Take 2.5 kg of fresh Sedum spectabile leaves, thoroughly chop them using a herb cutter, and then submerge them in 95% ethanol, with the liquid level about 3 cm above the leaves. Soak overnight at room temperature. The next day, rotary evaporate the soaking solution at 50°C until no ethanol residue remains. Take 1 / 10 of the extract as the ethanol extract sample, and evaporate it to dryness for storage.

[0056] The remaining extracts were then subjected to solvent extraction for separation: First, solvent was added at a volume ratio of extract:water:petroleum ether of 1:1:3. After thorough mixing, the mixture was allowed to stand and separate into layers. The upper petroleum ether phase was collected and evaporated to dryness at 50°C to obtain the petroleum ether fraction. Next, 3 times the volume of ethyl acetate was added to the lower aqueous phase for extraction. After thorough mixing, the mixture was allowed to stand and separate into layers. The upper ethyl acetate phase was collected and evaporated to dryness by rotary evaporation to obtain the ethyl acetate fraction. 3 times the volume of n-butanol was added to the remaining lower aqueous phase for extraction. After thorough mixing, the mixture was allowed to stand and separate into layers. The upper n-butanol phase was collected and evaporated to dryness by rotary evaporation at 50°C to obtain the n-butanol fraction. Finally, the remaining lower aqueous phase was rotary evaporated to dryness at 60°C to obtain the aqueous fraction. The masses of each fraction (ethanol, petroleum ether, ethyl acetate, and n-butanol fractions) were weighed and ultrasonically dispersed and suspended in ethanol to obtain corresponding 1 g / mL solutions. All samples were properly preserved for subsequent analysis of active ingredients and evaluation of bioactivity.

[0057] Comparative Example 2: Preparation of Kalanchoe extract and extracts from different parts of the plant. Take 2.5 kg of fresh Kalanchoe leaves, chop them using a herb cutter, and then immerse them in 95% ethanol, ensuring the liquid level is approximately 3 cm above the leaves. Extract using ultrasound-assisted extraction for 3 hours. After extraction, rotary evaporate the extract at 50°C until no ethanol residue remains, obtaining the crude extract. Take 1 / 10 of the extract volume as the ethanol extract fraction and retain it, then rotary dry for storage.

[0058] The remaining extracts were then subjected to solvent extraction for separation: First, solvent was added at a volume ratio of extract:water:petroleum ether of 1:1:3. After thorough mixing, the mixture was allowed to stand and separate into layers. The upper petroleum ether phase was collected and evaporated to dryness at 50°C to obtain the petroleum ether fraction. Next, 3 times the volume of ethyl acetate was added to the lower aqueous phase for extraction. After thorough mixing, the mixture was allowed to stand and separate into layers. The upper ethyl acetate phase was collected and evaporated to dryness by rotary evaporation to obtain the ethyl acetate fraction. 3 times the volume of n-butanol was added to the remaining lower aqueous phase for extraction. After thorough mixing, the mixture was allowed to stand and separate into layers. The upper n-butanol phase was collected and evaporated to dryness by rotary evaporation at 50°C to obtain the n-butanol fraction. Finally, the remaining lower aqueous phase was rotary evaporated to dryness at 60°C to obtain the aqueous fraction. Each of the above fractions (ethanol, petroleum ether, ethyl acetate, and n-butanol fractions) was ultrasonically dispersed and suspended in ethanol to obtain corresponding 1 g / mL extract solutions. All samples were properly preserved for subsequent analysis of active ingredients and evaluation of bioactivity.

[0059] Following the steps described above, active ingredients were extracted from 10 kg of Kalanchoe blossfeldiana leaves, 2.5 kg of Sedum spectabile, and 2.5 kg of Kalanchoe lanceolata using a solvent polar fraction extraction method. The yields of each fraction are shown in Table 1.

[0060] Table 1. Yields of extracts from different parts of Kalanchoe blossfeldiana, Sedum spectabile, and Kalanchoe pinnatifida.

[0061] Table 1 shows that the n-butanol fractions of Kalanchoe blossfeldiana, Sedum spectabile, and Kalanchoe lanceolata had the highest yields, reaching 0.16%, 0.05%, and 0.03%, respectively, indicating that n-butanol has the best extraction efficiency for the active ingredients of these three plants. In contrast, the ethyl acetate fractions of Kalanchoe blossfeldiana, Sedum spectabile, and Kalanchoe lanceolata had the lowest yields, at 0.05%, 0.01%, and 0.01%, respectively, suggesting that ethyl acetate solvent has a poor extraction effect on the corresponding plant materials. The results indicate that there are differences in the optimal extraction solvents for plant materials. Kalanchoe blossfeldiana, Sedum spectabile, and Kalanchoe lanceolata are more soluble in n-butanol, while the extraction efficiency in ethyl acetate solvent is lower, suggesting that the hydrophilic components of these plants may be more abundant than the lipophilic components.

[0062] Example 1: Effect of oral administration of medicinal plant ethanol extracts on mosquito mortality. To select the optimal plant materials, this embodiment uses three common Crassulaceae plants—Kalanchoe blossfeldiana (Kalanchoe thunbergii)—as raw materials. Kalanchoe blossfeldiana ), Sedum spectabile ( Hylotelephium spectabile ) and Kalanchoe ( Kalanchoe daigremontiana The ethanol extracts of three plant extracts were systematically evaluated for their mosquito-killing effects. The experiment tested the effectiveness of these extracts against Culex pipiens quinquefolius (a mosquito) via oral feeding. Culex quinquefasciatus ) and Aedes albopictus ( Aedes albopictus The kill rate of ).

[0063] Experimental Methods: The mosquito cages were labeled according to groups, with three replicates per group. Mosquitoes were anesthetized using carbon dioxide, and healthy individuals were selected on ice. The experiment was conducted in independent 30×30×25cm mosquito cages to test the killing effects of different plant ethanol extracts on different mosquito species. Different plant ethanol extracts (the Kalanchoe blossfeldiana ethanol extract sample prepared in Example 1, the Sedum spectabile ethanol extract sample prepared in Comparative Example 1, and the Kalanchoe blossfeldiana ethanol extract sample prepared in Comparative Example 2) were prepared into 1% (final concentration based on the mass percentage of the extract in the juice) ATSB solutions using fruit juice. The formulations for each treatment group were as follows: A. Blank (apple juice + 8% sugar water): 8g sucrose added to 100ml apple juice; B. Control (blank + ethanol): 8g sucrose added to 100ml ethanol; C. (Kalanchoe blossfeldiana + B): 1% Kalanchoe blossfeldiana ethanol extract was added to control B, i.e., 8g sucrose and 1ml of the 1g / mL extract solution obtained in Example 1 were added to 100ml ethanol. D (Kalanchoe + B): Add 1% Kalanchoe ethanol extract to control B, i.e., add 8g sucrose and 1ml of the 1g / mL extract solution obtained in Comparative Example 2 to 100ml ethanol. E (Sedum spectabile + B): Add 1% Sedum spectabile ethanol extract to control B, i.e., add 8g sucrose and 1ml of the 1g / mL extract solution obtained in Comparative Example 1 to 100ml ethanol. Release 30 experimental mosquitoes (half male and half female) that have emerged 3-10 days prior into each mosquito cage. After 8 hours of sugar deprivation treatment, place cotton balls soaked in 25mL of blank group solution (A), 25mL of control solution (B), 25mL of Kalanchoe ATSB experimental group solution (C), 25mL of Kalanchoe ATSB experimental group solution (D), and 25mL of Sedum spectabile ATSB experimental group solution (E) into the mosquito cages. Experimental conditions were controlled in a constant temperature and humidity environment of 26±1℃ and 70±5%, with each experiment repeated three times. Mosquito mortality rates were observed and recorded after 24 and 48 hours. Mosquito mortality was defined as follows: the mosquito's abdomen rolled upwards, its six legs twitched, and it was unable to roll over or crawl after being lightly touched with an insect stinger. The optimal plant ethanol extract was selected by comparing the killing effects of different plant ethanol extract ATSB solutions.

[0064] Experimental results: The results are as follows Figure 1 As shown, Figure 1 In (A), the mortality rate of male Culex quinquefolia in the Kalanchoe group (77.78%) was higher than that in the control group, Kalanchoe blossfeldiana group, and Sedum spectabile group (13.33%, 8.89%, 15.56%, respectively). Figure 1 In (B), compared with the control group, the mortality rates of female Culex pipiens quinquefolia in the Kalanchoe group were significantly different within 24 h and 48 h (22.22% vs 0.00%, p=0.0332; 33.33% vs 0.00%, p=0.0332); compared with the Kalanchoe group, the mortality rates of female Culex pipiens quinquefolia in the Kalanchoe group were significantly different within 24 h and 48 h (22.22% vs 0.00%, p=0.0332; 33.33% vs 0.00%, p=0.0332); compared with the Sedum group, the mortality rates of female Culex pipiens quinquefolia in the Kalanchoe group were significantly different within 24 h and 48 h (22.22% vs 0.00%, p=0.0332; 33.33% vs 0.00%, p=0.0332). Figure 1 In (C), the total mortality rate of Culex pipiens quinquefolia in the 48h Kalanchoe group (55.56%) was higher than that in the control group, Kalanchoe blossfeldiana group, and Sedum spectabile group (6.67%, 4.44%, 7.78%).

[0065] Figure 1 In (D), the mortality rate of male Aedes albopictus mosquitoes in the Kalanchoe group (73.33%) was higher than that in the control group, Kalanchoe blossfeldiana group, and Sedum spectabile group (10.00%, 23.33%, 10.00%, respectively). Figure 1 In (E), compared with the control group, the mortality rate of female Aedes albopictus mosquitoes in the Kalanchoe group was significantly different within 48 hours (36.67% vs 0.00%, p=0.0493), and compared with the Sedum spectabile group, the mortality rate of female Aedes albopictus mosquitoes in the Kalanchoe group was significantly different within 48 hours (36.67% vs 0.00%, p=0.0493). Figure 1 In (F), the total mosquito mortality rate of Aedes albopictus in the Kalanchoe group (55.00%) was higher than that in the control group, Kalanchoe blossfeldiana group, and Sedum spectabile group (5.00%, 13.33%, 5.00%, respectively). This suggests that, compared to the control group, Kalanchoe blossfeldiana extract may have a killing effect on Culex pipiens quinquefolius and Aedes albopictus after 48 hours. Based on the above experimental data, the present invention ultimately determined Kalanchoe blossfeldiana as the core plant material because its extracts showed stable and significant killing activity against a variety of major mosquito vectors.

[0066] Example 2: Effect of direct contact with Kalanchoe blossfeldiana ethanol extract on mosquito mortality rate Disposable food containers with a capacity of 350ml were labeled according to groups, with three replicates per group. Experimental mosquitoes were anesthetized with carbon dioxide and placed on ice to maintain their anesthesia. Twenty healthy mosquitoes (half male and half female) that had emerged 3-10 days prior, undergone 8 hours of sugar deprivation treatment, and were selected from the ice were then used. Using a pipette, 0.2 μL of different concentrations (0.1%, 1%, solvent: ethanol, dilution method as in Example 1; 1% is 100ml ethanol with 1mL of the 1g / mL ethanol extract solution obtained in Example 1) of the Kalanchoe blossfeldiana ethanol extract sample prepared in Example 1, 0.01g / L fipronil (as a positive control), and 95% ethanol (control group) were accurately pipetted onto the abdomen and dorsal plate of each mosquito. After application, the mosquitoes were transferred to disposable food containers and fed a normal 8% sucrose solution. The experiment was conducted in a constant temperature and humidity environment of 26±1℃ and 70±5%. The number of dead mosquitoes was observed and recorded every 12 hours, and the mortality rate was calculated. The criteria for determining mosquito mortality are: the mosquito's abdomen rolls upward, its six legs twitch, and it is unable to roll over or crawl after being lightly touched with an insect needle.

[0067] The results are as follows Figure 2 As shown, Figure 2 In (A), compared with the control group, the mortality rate of male Culex pipiens quinquefolia in the 0.1% Kalanchoe blossfeldiana group was higher than that in the control group (50.00% vs 18.33%), and the mortality rate of male Culex pipiens quinquefolia in the 1% Kalanchoe blossfeldiana extract group was higher than that in the control group (86.67% vs 18.33%). Figure 2 In (B), the mortality rate of male Culex pipiens quinquefolia in the 1% Kalanchoe extract group within 24 hours was significantly different from that in the control group (83.33% vs 3.33%, p=0.0313). Figure 2 In (C), the mortality rate of Culex pipiens quinquefolia in the 1% Kalanchoe blossfeldiana extract group was significantly different from that in the control group within 24 hours (86.67% vs 18.33%, p=0.0389). This indicates that the Kalanchoe blossfeldiana ethanol extract, when applied topically to the abdomen and dorsal surface of mosquitoes, has a significant killing effect within 24 hours, and this killing effect increases with increasing extract concentration.

[0068] Figure 2 In (DF), within 24 hours, the positive control group of fipronil showed significant killing effects on male and female Aedes albopictus mosquitoes and total mosquitoes. Figure 2 In (D), the mortality rate of male Aedes albopictus mosquitoes in the 0.1% Kalanchoe group was higher than that in the control group within 24 hours (96.67% vs 43.33%), and the mortality rate of male Aedes albopictus mosquitoes in the 1% Kalanchoe group was higher than that in the control group (86.67% vs 43.33%). Figure 2In (E), the mortality rate of female Aedes albopictus mosquitoes in the 0.1% Kalanchoe group was higher than that in the control group within 24 hours (56.67% vs 3.33%), and the mortality rate of female Aedes albopictus mosquitoes in the 1% Kalanchoe group was higher than that in the control group (86.67% vs 3.33%). Figure 2 In (F), the total mortality rate of Aedes albopictus in the 0.1% Kalanchoe blossfeldiana group was higher than that in the control group within 24 hours (76.67% vs 23.33%), and the total mortality rate of Aedes albopictus in the 1% Kalanchoe blossfeldiana group was higher than that in the control group (86.67% vs 23.33%). This suggests that Kalanchoe blossfeldiana extract may have the potential for direct contact killing of Aedes albopictus. However, within 24 hours, the direct contact killing effect of 1% Kalanchoe blossfeldiana extract on male Aedes albopictus was slightly lower than that of the 0.1% Kalanchoe blossfeldiana ethanol extract group. This may be due to operational reasons during the experiment. Preliminary overall data suggests that this killing ability may increase with increasing extract concentration.

[0069] The experimental results of Examples 1 and 2 demonstrate that the Kalanchoe extract has both gastric poisoning and contact killing effects, and the killing effect increases with increasing concentration, which provides a theoretical basis for its application in various scenarios (such as space spraying and residual treatment).

[0070] Example 3: Effect of oral administration of Kalanchoe blossfeldiana ethanol extract combined with insecticide on mosquito mortality. The chemical insecticide fipronil was combined with the ethanol extract of Kalanchoe blossfeldiana prepared in Example 1, mixed with apple juice, and sucrose, food coloring (Shanghai Dyestuff Research Institute Co., Ltd. Nantong Branch 10818001), and preservative (potassium sorbate, Shanghai Maclean Biochemical Technology Co., Ltd. 69023222) were added to prepare poisoned bait. The poisoned bait formulas for each treatment group are as follows: A: Control group (apple juice + 8% sugar water + 0.1% food coloring + 0.1% preservative + ethanol): 8g sucrose, 0.1ml food coloring, and 0.1mg preservative (dissolved in ethanol, volume negligible) were added to 100ml apple juice; B: 5×10 -4 g / L fipronil + A (using A as solvent, fipronil is added to achieve a final concentration of 5 × 10 g / L) -4 C: 0.1% Kalanchoe blossfeldiana ethanol extract + A (using 1000ml of A as solvent, add 1ml of the 1g / mL extract solution obtained in Preparation Example 1 to make the final concentration 0.1%); D: B + C (using 1000ml of A as solvent, add 1mL of the 1g / mL extract solution obtained in Preparation Example 1 to make the final concentration 0.1%, add 5×10 g / L). -4 g of fipronil, resulting in a final concentration of 5 × 10 -4The test insects were fed with a concentration of g / L, and the number of deaths was recorded every 24 hours. The mosquito species used were: 15 males and 15 females of Culex quinquefasciatus (30 in total), 15 males and 15 females of Aedes albopictus (30 in total), and 15 males and 15 females of Anopheles sinensis (30 in total). Experimental conditions were controlled in a constant temperature and humidity environment of 26±1℃ and 70±5%. All other procedures were the same as in Example 1.

[0071] The results are as follows Figure 3 As shown, Figure 3 In .3(A), within 48 hours, the killing rate of male Culex pipiens quinquefolius was higher in the 0.1% Kalanchoe blossfeldiana extract combined with fipronil group than in the fipronil group (86.67% vs 73.33%). Figure 3 In .3(B), within 24 hours, the kill rate of female Culex pipiens quinquefolius in the 0.1% Kalanchoe blossfeldiana extract combined with fipronil was significantly higher than that in the control group (16.67% vs 0.00%, p=0.0437), and within 48 hours, the kill rate of female Culex pipiens quinquefolius in the 0.1% Kalanchoe blossfeldiana extract combined with fipronil was significantly higher than that in the control group (53.33% vs 0.00%, p=0.0187). Figure 3 In .3(C), within 24h and 48h, the total kill rate of Culex pipiens quinquefolius in the combination of 0.1% Kalanchoe blossfeldiana extract and dinotefuran was significantly higher than that in the control group (28.33% vs 3.33%, p=0.0187; 70.00% vs 6.67%, p=0.0132). This indicates that the combination of Kalanchoe blossfeldiana extract and a single chemical insecticide can enhance the killing effect of chemical insecticides on Culex pipiens quinquefolius.

[0072] Figure 3In Figure 8(D), within 24 hours, the kill rate of male Aedes albopictus mosquitoes by the combination of 0.1% Kalanchoe blossfeldiana extract and dinotefuran was significantly higher than that of the control group (53.33% vs 10.00%, p=0.0298). In Figure 8(E), within 48 hours, the kill rate of female Aedes albopictus mosquitoes by the combination of 0.1% Kalanchoe blossfeldiana extract and dinotefuran was higher than that of the dinotefuran group (66.67% vs 63.33%). In Figure 8(F), within both 24 and 48 hours, the total kill rate of Aedes albopictus mosquitoes by the combination of 0.1% Kalanchoe blossfeldiana extract and dinotefuran was significantly higher than that of the control group (35.00% vs 6.67%, p=0.0298; 76.67% vs 23.33%, p=0.0361). Therefore, the combination of Kalanchoe blossfeldiana extract and a single chemical insecticide can enhance the killing effect of chemical insecticides on Aedes albopictus.

[0073] Figure 3 In .3 (G), within 48 hours, the killing rate of male Anopheles sinensis mosquitoes by the combination of 0.1% Kalanchoe blossfeldiana extract and dinotefuran was significantly higher than that of the control group (100.00% vs 46.67%, p=0.0164). Figure 3 In .3 (H), within 24 h, the killing rate of female Anopheles sinensis mosquitoes by the combination of 0.1% Kalanchoe blossfeldiana extract and dinotefuran was significantly higher than that by the control group (20.00% vs 0.00%, p=0.0448), the killing rate of female Anopheles sinensis mosquitoes by the combination of 0.1% Kalanchoe blossfeldiana extract and dinotefuran was significantly higher than that by the dinotefuran group (20.00% vs 0.00%, p=0.0448), and the killing rate of female Anopheles sinensis mosquitoes by the combination of 0.1% Kalanchoe blossfeldiana extract and dinotefuran was significantly higher than that by the 0.1% Kalanchoe blossfeldiana extract group (20.00% vs 0.00%, p=0.0448). Figure 3 In .3 (I), within 24h and 48h, the total kill rate of Anopheles sinensis mosquitoes by the combination of 0.1% Kalanchoe blossfeldiana extract and dinotefuran was significantly higher than that of the control group (38.33% vs 8.33%, p=0.0262; 63.33% vs 23.33%, p=0.0122). This indicates that the combination of Kalanchoe blossfeldiana extract and a single chemical insecticide can enhance the insecticidal effect of the chemical insecticide on mosquitoes and reduce the dosage of the chemical insecticide. Preliminary data suggest that the kill rate of Kalanchoe blossfeldiana extract on male mosquitoes is higher than that on female mosquitoes.

[0074] The results above show that even low concentrations of poisoned bait can produce significant differences, and it is expected that high concentrations can further increase the lethality rate.

[0075] Example 4: The effect of a combination of Kalanchoe blossfeldiana ethanol extract and insecticide on mosquito mortality in a simulated field. To verify the effectiveness of this technology in a more realistic application environment, this embodiment conducted a simulated field experiment in a large mosquito net (90×110×190cm). Two hundred Culex pipiens quinquefolius mosquitoes (half male and half female) were released. Five sponges (5×5×2 cm each) were placed in the mosquito net to ensure the mosquitoes could ingest the bait. The mosquitoes were fed the bait, and the killing effect of different treatment groups was tested. The formulations for each treatment group are shown below. Figure 4 The preparation process was the same as in Example 3. The food coloring and preservatives were also the same as in Example 3. The number of deaths was observed and recorded every 24 hours.

[0076] Experimental results are as follows Figure 4 As shown, Figure 4 In (A), compared with the control group, at 48h, 1×10 -3 The mortality rate of male Culex pipiens quinquefolius was significantly different in the g / L fipronil group (68.67% vs 36.00%, p=0.0174), with a mortality rate of 1×10⁻⁶. -2 The mortality rates of male Culex pipiens quinquefolius were significantly different in the g / L fipronil group (84.00% vs 36.00%, p=0.0012), in the 0.1% Kalanchoe blossfeldiana group (78.33% vs 36.00%, p=0.0031), and in the Kalanchoe blossfeldiana compound group (92.67% vs 36.00%, p=0.0003).

[0077] Figure 4 In (B), compared with the control group, at 48h, 1×10 -2 The mortality rates of female Culex pipiens quinquefolius were significantly different in the 1 g / L fipronil group (78.67% vs 21.00%, p=0.0022), 60.67% vs 21.00% in the 0.1% Kalanchoe blossfeldiana group (p=0.0260), and 86.33% vs 21.00% in the Kalanchoe blossfeldiana compound group (p=0.0008). This was significantly different from the 1×10 -3 Compared with the g / L fipronil group, the mortality rate of female Culex quinquefolia in the Kalanchoe blossfeldiana compound group was significantly different at 48h (86.33% vs 47.33%, p=0.0287).

[0078] Figure 4 In (C), compared with the control group, 1×10 -2 The total mortality rate of Culex pipiens quinquefolius in the 1 g / L fipronil group was significantly different (45.33% vs 19.00%, p=0.0133), and the total mortality rate of Culex pipiens quinquefolius in the Kalanchoe blossfeldiana compound group was significantly different (51.67% vs 19.00%, p=0.0031). Compared with the control group, the total mortality rate of Culex pipiens quinquefolius in the 1 g / L fipronil group was significantly lower at 48 h. -3The total mortality rate of Culex pipiens quinquefolius was significantly different in the g / L fipronil group (58.00% vs 28.50%, p=0.0284), with a mortality rate of 1×10⁻⁶. -2 The total mortality rate of Culex pipiens quinquefolius was significantly different in the 1 g / L fipronil group (81.33% vs 28.50%, p=0.0005), in the 0.1% Kalanchoe blossfeldiana group (69.50% vs 28.50%, p=0.0034), and in the Kalanchoe blossfeldiana compound group (89.50% vs 28.50%, p=0.0001); compared with 1×10 -3 Compared to the g / L fipronil group, the mortality rate of female Culex pipiens quinquefolius in the Kalanchoe blossfeldiana compound group was significantly different at 48 h (89.50% vs 58.00%, p=0.0194). This indicates that Kalanchoe blossfeldiana extract has the potential to kill Culex pipiens quinquefolius in larger spaces, and that when combined with chemical insecticides, it significantly enhances the killing effect of single chemical insecticides on Culex pipiens quinquefolius.

[0079] Example 5: Effects of Kalanchoe blossfeldiana ethanol extract on mosquito mortality rate using solvents of different polarities To determine the main enrichment sites of mosquito-killing activity in Kalanchoe blossfeldiana, this example conducted oral toxicity tests on extracts with different polar solvents (petroleum ether, ethyl acetate, and n-butanol fractions). The experiment used Culex pipiens quinquefolius as the target mosquito.

[0080] The experimental mosquito cages were labeled according to groups, with each group having three replicates. The mosquitoes were anesthetized using carbon dioxide, and healthy individuals were selected on ice. The experiment was conducted in independent mosquito cages (30cm long, 30cm wide, and 25cm high) to test the killing effect of different polar solvent extracts of Kalanchoe blossfeldiana on different mosquito species. Thirty mosquitoes (half male, half female) aged 3-10 days after emergence were released into each mosquito cage. After 8 hours of sugar-free treatment, cotton balls soaked in either 25mL of the experimental group solution (ethanol extract and petroleum ether, ethyl acetate, or n-butanol fractions) or 25mL of the control group solution were placed in the mosquito cages. The specific groups are as follows: Blank group (A): apple juice + 0.01% food coloring + 0.01% preservative; Control group (B): ethanol + 0.01% food coloring + 0.01% preservative; Experimental group includes 1% Kalanchoe blossfeldiana ethanol extract / 1% Kalanchoe blossfeldiana petroleum ether extract / 1% Kalanchoe blossfeldiana ethyl acetate extract / 1% Kalanchoe blossfeldiana n-butanol extract + ethanol + 0.01% food coloring + 0.01% preservative; The preparation process is the same as in Example 3. The experimental conditions are controlled in a constant temperature and humidity environment of 26±1℃ and 70±5%, and each group of experiments is repeated three times. The mortality rate of mosquitoes is observed and recorded after 24h and 48h. The criteria for judging the death of mosquitoes are: the mosquito's abdomen rolls upward, its six legs twitch, and it cannot roll over or crawl after being lightly touched with an insect needle. By comparing the killing effect of ATSB solutions of different plant polar parts extracts, the best plant polar part extract is screened.

[0081] Experimental results are as follows Figure 5 As shown in the figure, within 48 hours, the total mortality rate of Culex pipiens quinquefolia extract group was as high as 90.00%, significantly higher than that of the control group (6.67%, p=0.0086), petroleum ether group (16.67%), and n-butanol group (55.56%). For female mosquitoes, the 48-hour mortality rate of the ethyl acetate group also reached 80.00%, significantly higher than that of the control group (0.00%, p=0.0204). The results show that the ethyl acetate fraction had the most significant killing effect. This result clearly indicates that the substances with highly effective mosquito-killing activity in Kalanchoe blossfeldiana are mainly moderately polar compounds that can be effectively extracted by ethyl acetate.

[0082] The present invention has been described in detail above. Those skilled in the art will recognize that the invention can be practiced in a wide range of ways with equivalent parameters, concentrations, and conditions without departing from its spirit and scope. While specific embodiments have been provided, it should be understood that further modifications can be made to the invention. In summary, according to the principles of the invention, this application is intended to include any changes, uses, or improvements to the invention, including modifications made using conventional techniques known in the art that depart from the scope disclosed herein.

Claims

1. Use of ethanol extract of leaves of Euphorbia milii in killing mosquitoes or preparing a product for killing mosquitoes.

2. Use according to claim 1, characterized in that: The mosquitoes are selected from one or more of Culex quinquefasciatus, Aedes albopictus, Anopheles sinensis; and / or, The mosquitoes are male mosquitoes and / or female mosquitoes; and / or, The killing is by feeding and / or contact killing.

3. A mosquito control composition, characterized by, The composition comprises a sugar source, the ethanol extract of leaves of Euphorbia milii, and a solvent.

4. The mosquito control composition according to claim 3, wherein: The sugar source is selected from one or more of sucrose, honey, fructose, glucose; and / or, The concentration of the sugar source in the composition is 5% to 15% by mass volume percentage; and / or, The concentration of the ethanol extract of leaves of Euphorbia milii in the composition is 0.1% to 1% by mass volume percentage; and / or, The solvent is ethanol.

5. A mosquito control composition comprising, wherein, The composition comprises a sugar source, the ethanol extract of leaves of Euphorbia milii, dinotefuran, and fruit juice.

6. The mosquito control composition of claim 5, wherein: The sugar source is selected from one or more of sucrose, honey, fructose, glucose; and / or, The concentration of the sugar source in the composition is 5% to 15% by mass volume percentage; and / or, The concentration of the ethanol extract of leaves of Euphorbia milii in the composition is 0.1% to 1% by mass volume percentage; and / or, The concentration of the dinotefuran in the complex composition is 1 x 10 -4 g / L ~ 1 x 10 -2 g / L; and / or, The mass ratio of the ethanol extract of Euphorbia milii to dinotefuran is (2000 to 10): 1; and / or, The fruit juice is apple juice or pear juice; and / or, The composition further comprises an edible colorant and / or a preservative; further, the edible colorant is present in the composition at a volume percentage of 0.01% to 0.5%; the preservative is present in the composition at a mass volume percentage of 0.01% to 0.5%; further, the preservative can be potassium sorbate.

7. Use of an ethyl acetate extract of an ethanol extract of a leaf of Gypsophila paniculata in killing mosquitoes or in the preparation of a product for killing mosquitoes; wherein, The preparation of the ethyl acetate extract of the ethanol extract of leaves of Euphorbia milii comprises: 1) adding water and petroleum ether to the ethanol extract of leaves of Euphorbia milii, thoroughly mixing, and then allowing to stand to separate into layers, and collecting the lower aqueous phase; 2) adding ethyl acetate to the aqueous phase to perform extraction, thoroughly mixing, and then allowing to stand to separate into layers, and collecting the upper ethyl acetate phase, and evaporating to dryness to obtain the ethyl acetate extract.

8. A mosquito control composition, characterized by, The composition comprises the ethyl acetate extract of the ethanol extract of leaves of Euphorbia milii and a solvent; wherein the preparation of the ethyl acetate extract of the ethanol extract of leaves of Euphorbia milii comprises: 1) adding water and petroleum ether to the ethanol extract of leaves of Euphorbia milii, thoroughly mixing, and then allowing to stand to separate into layers, and collecting the lower aqueous phase; 2) adding ethyl acetate to the aqueous phase to perform extraction, thoroughly mixing, and then allowing to stand to separate into layers, and collecting the upper ethyl acetate phase, and evaporating to dryness to obtain the ethyl acetate extract.

9. The mosquito control composition of claim 8, wherein: The concentration of the ethanol extract of leaves of Euphorbia milii in the composition is 0.1% to 1% by mass volume percentage; and / or, The solvent is ethanol; and / or, The composition further comprises an edible colorant and / or a preservative.

10. A method of mosquito control, comprising, The method comprises the step of deploying the mosquito control composition of claim 3 or 4, or the mosquito control composition of claim 5 or 6, or the mosquito control composition of claim 8 or 9, to a mosquito breeding site or habitat to kill mosquitoes.

Images