Application of cinnamyl aldehyde and composition containing cinnamyl aldehyde in prevention and treatment of parasitic ticks

Abstract

The invention relates to cinnamyl aldehyde and application of a composition containing the cinnamyl aldehyde to prevention and treatment of parasitic ticks. The present invention relates to the field of biocides. Cinnamaldehyde is found for the first time to have insecticidal activity on ticks, especially hard ticks, and is safe to the host protaetia brevitarsis adults of the ticks under a certain dosage, so that cinnamaldehyde can be used as the insecticide for the ticks. Furthermore, although the vermiculite has no insecticidal activity on the ticks, when the cinnamyl aldehyde is added into the vermiculite and the composition of the cinnamyl aldehyde and the vermiculite is used for preventing and treating the ticks, the vermiculite can improve the insecticidal effect of the cinnamyl aldehyde. And in combination with the soil diving habit of the adult protaetia brevitarsis, vermiculite coincidence provides a shielding place for the protaetia brevitarsis, and operability is provided for preventing and treating parasitic ticks.

Description

Technical Field

[0001] The invention relates to the field of biocides, and in particular to application of cinnamaldehyde and a composition containing the cinnamaldehyde in preventing and controlling parasitic ticks. Background Art

[0002] White-starred flower beetle ( Protaetia brevitarsis , PB) is a soil-dwelling insect belonging to the class Insecta, order Coleoptera, family Cetoniidae, genus Protaetia. Also known as white-striped copper beetle, white-star flower beetle, white-star beetle, copper beetle, it undergoes complete metamorphosis. Under natural conditions, adults generally occur on economic crops with a sweet smell, while larvae exist in environments with high organic matter content such as rotten straw, fallen wood, and livestock and poultry manure. The white-star flower beetle is widely distributed in Northeast China, North China, several eastern provinces along the southeast coast, and parts of the northwest, and occurs more frequently in countries and regions such as Japan, North Korea, Mongolia, and Russia.

[0003] The larvae of the white-star flower beetle are rich in nutrients, containing a large amount of protein, peptides, fatty acids, sugars, alkaloids, antimicrobial peptides, vitamins and a small amount of mineral elements. Therefore, the larvae of the white-star flower beetle are widely used and have played a role in many fields such as medicine, food, aquaculture, agriculture and forestry, and resource environmental protection. At the same time, the larvae of the white-star flower beetle are a good source of protein. The protein content of the larvae of the white-star flower beetle is 57.86±0.01% of the dry weight, which is similar to the silkworms (55% to 76%) and locusts (68% to 78%) among the insects currently permitted for consumption. And it is similar to the protein content in eggs, 1.80 times that of milk and 2.34 times that of pork. It can be used as an important source of new protein resource food, especially as a protein resource for animal feed. In 2018, the larvae of the white-star flower beetle were registered as food ingredients by the Food and Drug Administration (Korea Food and Drug Safety). With the deepening of research, the value of the white-star flower beetle in agriculture has gradually been discovered, and its comprehensive application in agriculture has become more and more extensive.

[0004] The larvae of the white-spotted flower beetle are commonly used medicinal materials. They are used in the formulation of the preparations of Daman Chong Wan, Ganqi Wan, Shanzhu Yu Wan, Tianjiangzi Wan, etc., and in the prescriptions of Dahuang Chong Teng Wan, Dahuang Chong Teng Capsules, and Dahuang Chong Teng Tablets. In Korea, the larvae of the white-spotted flower beetle are a traditional medicinal material in Korean medicine. According to the Dongyi Bogam, the larvae of the white-spotted flower beetle can be ground into powder and used as medicine to treat liver cancer, cirrhosis, hepatitis, relieve fatigue, and regulate menstrual disorders. It also has certain therapeutic effects on diseases such as vision loss, cataracts, postpartum wind, ulcers, stomatitis, and stroke, especially for liver disease. It has a significant therapeutic or relieving effect, so it is often used in Korean medicine and folk remedies. In addition, the larvae of the white-spotted flower beetle contain a large number of active substances related to health. For example, the protein isolate of the larvae of the white-spotted flower beetle can also be used as a potential antioxidant biomaterial for bone tissue engineering applications.

[0005] The feces of the larvae of the white-spotted flower beetle can be developed into nutrient-rich organic fertilizer, which will promote the development of the agricultural fertilizer field.

[0006] During the breeding process of white-starred flower beetles, adults are easily parasitic by ticks, which can lead to the death of the white-starred flower beetle group in severe cases, which is a major problem that plagues the breeding of white-starred flower beetles. These parasitic ticks mostly parasitize externally, and mainly parasitize on the base of the host's feet, the connection between the chest and abdomen, and the connection between the head and thorax. They suck the body fluids and hemolymph of adults and larvae to complete their growth and reproduction, which seriously threatens the growth, development and health of white-starred flower beetles. However, there are currently no products and technologies for the prevention and control of parasitic ticks on white-starred flower beetles, which restricts the development of the white-starred flower beetle industry. Summary of the invention

[0007] One of the present inventions provides the use of cinnamaldehyde in preventing and controlling ticks.

[0008] In a specific embodiment, the tick is a species of Ixodes.

[0009] In a specific embodiment, the mitochondrial sequence of the tick is shown as SEQ ID No. 1.

[0010] In a specific embodiment, the tick also has the following characteristics: its developmental life cycle goes through four stages: egg, larva, nymph and adult; wherein, its eggs are ellipsoidal or spherical, off-white or milky white, and are translucent as a whole; when the larva is first hatched, the insect body is white and translucent, the larva has three pairs of legs, and the nymph and adult tick have four pairs of legs; the length-to-width ratio of the body of the larva and early nymph is about 2:1, and is long and oval; the length-to-width ratio of the body of the late nymph and adult tick is about 1.5:1, and is oval; it undergoes two moltings from larva to nymph and from nymph to adult; the adult tick is about 0.65 mm long, about 0.40 mm wide, oval, reddish brown or grayish brown in color, and has piercing-sucking mouthparts.

[0011] In a specific embodiment, it takes about 40 to 45 days for the tick to develop from an egg to an adult tick.

[0012] In a specific embodiment, the tick undergoes two molts, from larva to nymph and from nymph to adult.

[0013] In a specific embodiment, the tick is divided into a pseudo-head and a body, without a head, thorax and abdomen, and the ventral surface is flat, and its pseudo-head can be seen from the dorsal view; the body is hard and smooth, and there are about 12 50-micron-long hairs on the dorsal side of the outer edge of the tail end.

[0014] In a specific embodiment, the tick has an overall strong movement ability, which gradually increases as the tick matures. The movement speed of a young tick is about 0.28 cm / s, and the movement speed of an adult tick is about 0.46 cm / s.

[0015] The second aspect of the present invention provides a composition comprising cinnamaldehyde and vermiculite.

[0016] In a specific embodiment, based on 100% by mass of the composition, the content of cinnamaldehyde is above 0.4 mg / g.

[0017] In a specific embodiment, based on 100% by mass of the composition, the content of cinnamaldehyde is greater than 0.4 mg / g and less than 4 mg / g.

[0018] In a specific embodiment, based on 100% by mass of the composition, the content of cinnamaldehyde is greater than 0.88 mg / g and less than 2.63 mg / g.

[0019] The third aspect of the present invention provides the use of the composition according to any one of the second aspects of the present invention in controlling ticks.

[0020] In a specific embodiment, the tick is Ixodes.

[0021] In a specific embodiment, the mitochondrial sequence of the tick is shown as SEQ ID No. 1.

[0022] In a specific embodiment, the tick also has the following characteristics: its developmental life cycle goes through four stages: egg, larva, nymph and adult; wherein, its eggs are ellipsoidal or spherical, off-white or milky white, and are translucent as a whole; when the larva is first hatched, the insect body is white and translucent, the larva has three pairs of legs, and the nymph and adult tick have four pairs of legs; the length-to-width ratio of the body of the larva and early nymph is about 2:1, and is long and oval; the length-to-width ratio of the body of the late nymph and adult tick is about 1.5:1, and is oval; it undergoes two moltings from larva to nymph and from nymph to adult; the adult tick is about 0.65 mm long, about 0.40 mm wide, oval, reddish brown or grayish brown in color, and has piercing-sucking mouthparts.

[0023] In a specific embodiment, it takes about 40 to 45 days for the tick to develop from an egg to an adult tick.

[0024] In a specific embodiment, the tick undergoes two molts, from larva to nymph and from nymph to adult.

[0025] In a specific embodiment, the tick is divided into a pseudo-head and a body, without a head, thorax and abdomen, and the ventral surface is flat, and its pseudo-head can be seen from the dorsal view; the body is hard and smooth, and there are about 12 50-micron-long hairs on the dorsal side of the outer edge of the tail end.

[0026] In a specific embodiment, the tick has an overall strong movement ability, which gradually increases as the tick matures. The movement speed of a young tick is about 0.28 cm / s, and the movement speed of an adult tick is about 0.46 cm / s.

[0027] Beneficial effects of the present invention: The present invention first discovered that cinnamaldehyde has insecticidal activity against ticks, especially hard ticks (Ixodes), and it is safe to the host of the ticks, the adult white-spotted flower beetle, at a certain dose. Therefore, cinnamaldehyde can be used as an insecticide for ticks. However, when the contact method is used to kill ticks parasitizing on the body of the adult white-spotted flower beetle, since the parasitism of ticks mostly occurs at the base of the foot, the connection between the thorax and abdomen, and the connection between the head and thorax of the adult white-spotted flower beetle, the spray method is difficult to reach these parasitized parts of the adult white-spotted flower beetle, so it is difficult to achieve a tick-killing effect equivalent to that under non-parasitic conditions in actual operation; fumigation to kill ticks requires the airtightness of the tick-killing space, otherwise the concentration of cinnamaldehyde in the space cannot be guaranteed, but the oxygen supply of the adult white-spotted flower beetle cannot be guaranteed in such a closed space, affecting its health. Furthermore, the present invention also found that although vermiculite has no insecticidal activity against ticks, when cinnamaldehyde is added to vermiculite and a composition of cinnamaldehyde and vermiculite is used to control ticks, vermiculite can improve the insecticidal effect of cinnamaldehyde. In combination with the fact that the adult white-spotted flower beetle has the habit of lurking in the soil, vermiculite coincidentally provides a shelter for it, providing operability for controlling parasitic ticks. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 The various stages of the tick life cycle are shown, where A is the egg, B is the larva, C is the early nymph, D is the late nymph, E is the adult, and F is the molt.

[0029] Figure 2 Shows the parasitism rate of ticks on adults of the white-spotted flower beetle.

[0030] Figure 3 The comparison results of the mitochondrial sequence of the test tick of the present invention and the mitochondrial sequence of known tick species are shown.

[0031] Figure 4 The LC values ​​of cinnamaldehyde in Example 4 against ticks and cinnamaldehyde in Example 5 against adults of white-spotted flower beetles are shown. 50 Comparison of insecticidal activity. DETAILED DESCRIPTION

[0032] The above contents of the present invention are further described in detail below in the form of preferred implementation cases, but they do not constitute a limitation to the present invention.

[0033] Unless otherwise specified, the reagents in the embodiments of the present invention can be purchased through commercial channels.

[0034] Test insects: White-spotted flower beetle (WSFC) adults, collected in the insect rearing room of the Langfang pilot base of the Chinese Academy of Agricultural Sciences. After the eggs hatched, the larvae grew to the third instar and pupated, and then emerged as adults.

[0035] Test ticks: The parasitic ticks of the present invention were separated from adults of white-spotted flower beetles raised in the Langfang pilot base of the Chinese Academy of Agricultural Sciences and raised in vitro.

[0036] Tick ​​feed: Crush chicken bones and bone marrow into powder, add an equal amount of pure water to make chicken bone homogenate. Take the total mass of chicken bone homogenate as 100%, add 0.1wt% potassium sorbate as a preservative, 1.0wt% jelly powder to the chicken bone homogenate, sterilize at 121℃ for 15 min, pour into a mold to cool, solidify, shape, and store in a refrigerator at 4℃.

[0037] Tick ​​feeding: Use naturally fermented edible fungus residue with a humidity of 50% as the feeding matrix, take 200 g of the feeding matrix, and put an appropriate amount of tick feed into it. Select ticks from the body surface of adult white-spotted flower beetles parasitized with ticks and inoculate them into the tick feed. Place them in an artificial climate box, set the temperature to 26±1 degrees Celsius, the humidity to 50±5%, and the photoperiod to 16L:8D for cultivation. According to the experimental requirements, collect tick eggs, nymphs, and adults from the matrix for testing.

[0038] Inoculating ticks into adult white-spotted flower beetles: Healthy adult white-spotted flower beetles were placed into a feeding matrix with ticks and fed for 24 hours to complete the inoculation of adult white-spotted flower beetles with ticks for further testing.

[0039] Cinnamaldehyde, density 1.05 g / ml, was purchased from Sangon Biotech (Shanghai) Co., Ltd.

[0040] The Magnetic Universal Genomic DNA Kit used to extract genomic DNA was purchased from TIANGEN.

[0041] The Rapid Plus DNA Lib Prep Kit for Illumina (RK20208) for tick DNA sequencing was purchased from Abclonal. Example 1: Identification of tick populations parasitic on the white-spotted flower beetle

[0042] Pour a 1wt% agar plate as a base in a 35 mm culture dish. After the agar plate solidifies, place 3.0 g of tick feed in the center and inoculate 30 adult ticks separated from adult white-spotted flower beetles and artificially cultured. The breeding conditions are an ambient temperature of 26±1 degrees Celsius, a humidity of 50±5%, and a photoperiod of 16L:8D to observe the developmental life history of parasitic ticks.

[0043] The biological characteristics of the parasitic tick involved in the present invention are analyzed as follows: through the above artificial culture observation, it is determined that the developmental life history of the parasitic tick goes through four stages: egg, larvae, nymph and adult tick. Among them, it takes about 40 to 45 days for the egg to develop into an adult tick. The egg is ellipsoidal or spherical, off-white or milky white, and is translucent as a whole. When the larvae are first hatched, the insect body is white and translucent, the larvae have three pairs of legs, and the nymphs and adult ticks have four pairs of legs. The length-to-width ratio of the body of the larvae and the early nymphs is about 2:1, and they are oblong, and the length-to-width ratio of the body of the late nymphs and adult ticks is about 1.5:1, and they are oval. After two molts from larvae to nymphs and nymphs to adult ticks, the body color is milky white within a few days after molting. The adult tick is about 0.65 mm long, about 0.40 mm wide, oval, reddish brown or grayish brown in color, and has a piercing-sucking mouthpart. The tick is divided into two parts: the pseudo-head and the body. There is no distinction between the head, thorax and abdomen. The ventral side is flat, and the pseudo-head can be seen from the back. The body is hard and smooth, and there are about 12 50-micron-long hairs on the dorsal side of the outer edge of the tail. The tick has strong overall movement ability, which gradually increases as the insect matures. The movement speed of the larval tick is about 0.28 cm / s, and the movement speed of the adult tick is about 0.46 cm / s. Figure 1 The various stages of the tick life cycle are shown.

[0044] Observation shows that under natural conditions, the larvae, nymphs and adult ticks of the parasitic ticks involved in the present invention all feed on the host, and their bodies swell slightly after feeding. Ticks are photophobic and gregarious, and prefer humid environments. When encountering external physical stimulation, they curl up their feet and pretend to be dead. Parasitic ticks lay eggs in shaded places (such as under stones, soil blocks, and in wall cracks). Male and female mate, and the female tick falls to the ground and lays eggs after feeding.

[0045] Verification of the parasitic ability of parasitic ticks: Healthy individuals of white-spotted flower beetles were taken from the indoor breeding room of white-spotted flower beetles at the Langfang pilot base of the Chinese Academy of Agricultural Sciences. Two 35 mm culture dishes were placed mouth to mouth, one of which had a water agar bottom to facilitate tick inoculation. The junction of the two dishes was connected with a breathable tape as a parasitic tick rearing device. After 20 ticks of the same age were inoculated into the parasitic tick rearing device for 12 h of starvation, one WSFC adult was put in, and the parasitic ticks were cultured in a standardized culture environment of 26±1 degrees Celsius, 50±5% humidity, and 16L:8D light cycle as a standard parasitic model. The parasitic behavior of ticks was observed and recorded at 3, 6, 12, 24, 48 and 72 h after the start of the experiment, and the parasitic rate was calculated. The experiment was repeated 7 times. Parasitic rate (%) = 100 × number of parasitic host ticks / total number of ticks, the results are shown in Figure 2 .

[0046] Through observation, it was found that tick parasitism mostly occurred at the base of the feet, the connection between the thorax and abdomen, and the connection between the head and thorax of the white-spotted flower beetle, which is similar to what occurs during breeding, indicating that the ticks separated by the present invention are the main harmful population.

[0047] Figure 2 The results showed that the parasitism rates of larval ticks, nymphal ticks and adult ticks increased with the increase of treatment time. Within 6 h of treatment, the parasitism rates of the three were relatively low, at 17.86%, 29.29% and 23.57%, respectively, and there was no significant difference with the parasitism rates at 3 h of treatment ( P >0.05). The parasitism rate gradually stabilized after 48 hours of treatment, and the parasitism rates at 72 hours of treatment were 92.14%, 87.86% and 85.71%, respectively. Among the treatments of tick infestation at different ages, the parasitism rate at 12 hours of treatment was the most significant, with the lowest parasitism rate of larval ticks at 25.00%; the highest parasitism rate of adult ticks was 63.29%. The parasitism rate of larval ticks increased the most during the 12 and 24 hours of treatment, and the parasitism rates of the two were extremely significantly different ( P <0.01). It can be seen that all developmental stages of parasitic ticks parasitize and harm the white-spotted flower beetle. Among them, the parasitic behavior of larval ticks and nymph ticks is more active than that of adult ticks. Nymph ticks infect the fastest, and the parasitic rate of larval ticks is the highest under long-term treatment. From the perspective of infection speed, nymph ticks have the fastest infection speed, and the parasitic rate exceeds 50% after only 12 hours of treatment; however, since adult ticks are relatively larger, they are more convenient to handle. Therefore, subsequent experiments use adult ticks as experimental objects.

[0048] Analysis of the species of parasitic ticks on the white-spotted flower beetle: The whole genome of the parasitic ticks cultured under standardized in vitro conditions was extracted and subjected to high-throughput sequencing. The Rapid Plus DNA Lib Prep Kit for Illumina (RK20208) was used to construct a sample library. The mitochondria were assembled and annotated using Mitoz software to obtain a complete 14438 bp circular mitochondrial sequence (as shown in SEQ ID No. 1), with a total of 13 CDS, 2 rRNAs (I-rRNA and s-rRNA), and 22 tRNAs. The sequence was analyzed using the BLAST online platform of the National Center for Biotechnology Information of the United States. The results are shown in Figure 3 . Figure 3 The results of mitochondrial sequence comparison showed that the tested ticks and Ixodes rubicundus The overall comparison score is the highest, but compared with Ixodes The sequence alignment similarities of the tested ticks were all lower than 80%, so the tested tick was a new species of the genus Ixodes. Example 2: Determination of contact activity of the agent

[0049] The contact activity was determined by the Potter spray method, and the tick mortality rates at 12 h and 24 h were recorded to calculate the contact LC 50 .

[0050] Using 1% Tween-80 and 10% anhydrous ethanol aqueous solution as solvents, cinnamaldehyde was added dropwise to the solvent in a constant temperature water bath at 70 degrees Celsius to prepare seven drug solutions with different gradient concentrations.

[0051] 10 adult ticks were directly placed in a parasitic tick rearing device with a diameter of 35 mm, and sprayed under a Potter spray tower (pressure 100 kPa, spray volume 100 μL each time, sedimentation 2 min), and the experiment was repeated 7 times. The solvent treatment was used as a negative control, and then the ticks were placed in a 26°C constant temperature incubator for rearing. The results were investigated after 12 and 24 h, and the number of dead ticks was recorded, the corrected mortality rate was calculated, and the LC was calculated based on the corrected mortality rate at each concentration. 50 The experiment was repeated 5 times. Among them, the insects that did not respond to the brush touch, could not crawl normally, or had wrinkles, discoloration, deformation, and internal contents leaked out were judged as dead. Mortality rate (%) = number of dead insects / total number of insects × 100; Corrected mortality rate (%) = 100 × (mortality rate of treatment group-mortality rate of control group) / (1-mortality rate of control group).

[0052] The results showed that cinnamaldehyde had significant contact toxicity to the tested ticks under the spray treatment, and the contact toxicity increased with time. 50 The LC value of the sample was 5.50 g / L after 24 h treatment.50 It is 4.27 g / L. Example 3: Determination of fumigation activity of the agent

[0053] The indoor fumigation activity was determined by the closed petri dish fumigation method, and the tick mortality rates at 12 h and 24 h were recorded to calculate the fumigation LC 50 .

[0054] Cut a strip of filter paper (length 1.0 cm × width 0.5 cm), sterilize it, and use double-sided tape to fix both ends of the filter paper strip on top of the parasitic tick breeding device with a diameter of 35 mm, so that the filter paper strip is fully suspended in the space of the culture dish to facilitate the volatilization of cinnamaldehyde. Among them, the concentration of cinnamaldehyde is calculated based on the ratio of the mass of cinnamaldehyde added to the filter paper strip to the spatial volume of the culture dish. Inoculate 10 adult ticks in the culture dish, then drop different amounts of cinnamaldehyde on the middle of the filter paper strip, quickly seal it with a sealing film, and place it in a constant temperature incubator at 26 degrees Celsius for breeding. Investigate the results after 12 and 24 hours, record the number of dead ticks, calculate the corrected mortality rate, and calculate the LC based on the corrected mortality rate at each concentration. 50 , the experiment was repeated 5 times.

[0055] The results showed that cinnamaldehyde had significant fumigation toxicity to the tested ticks under fumigation treatment, and the toxicity increased with time. 50 The LC value after 24 h treatment was 320.6 mg / L. 50 It is 238.5 mg / L. Example 4: Determination of the activity of cinnamaldehyde and vermiculite composition against ticks

[0056] The toxicity of the combination of cinnamaldehyde and vermiculite to ticks was tested. The tick mortality rates at 12 and 24 h were recorded, and the LC 50 .

[0057] Using 1% Tween-80 and 10% anhydrous ethanol aqueous solution as solvents, cinnamaldehyde was added dropwise into the solvents in a constant temperature water bath at 70 degrees Celsius to prepare 10 ml of different doses of drug solution.

[0058] Prepare 20 g sterilized vermiculite as the matrix, add 10 ml of the solution at each dose, stir evenly with a glass rod, and place for 30 min to allow the vermiculite to fully absorb the free solution to obtain different ratios of cinnamaldehyde and vermiculite compositions. Fill 1.0 g of each dose of cinnamaldehyde and vermiculite composition into the parasitic tick breeding device, and inoculate 10 adult ticks into it. Record the number of dead ticks after 12 and 24 hours, respectively. Repeat the experiment 5 times. Among them, only adding 10 ml of solvent to 20 g of vermiculite was used as a negative control without cinnamaldehyde.

[0059] The results showed that the negative control (vermiculite) had no insecticidal effect on ticks. The adjusted mortality was calculated based on the negative control and the LC was calculated based on the adjusted mortality of cinnamaldehyde at each dose. 50 The results showed that in the combination of cinnamaldehyde and vermiculite, the higher the cinnamaldehyde content, the higher the insecticidal toxicity to the tested ticks; the LC 50 is 2.76 mg / g (the unit is the mass ratio of cinnamaldehyde to the mass ratio of cinnamaldehyde and vermiculite), and the LC 50 It is 0.44 mg / g (this unit is the ratio of the mass of cinnamaldehyde to the mass of the combination of cinnamaldehyde and vermiculite). Example 5: Determination of the activity of cinnamaldehyde and vermiculite composition against adults of white-spotted flower beetles

[0060] The toxicity of the combination of cinnamaldehyde and vermiculite to healthy adults of the white-star flower beetle was tested, and the mortality of the adults of the white-star flower beetle was recorded at 12 h and 24 h, and the LC of the adults of the white-star flower beetle was calculated. 50 .

[0061] Using 1% Tween-80 and 10% anhydrous ethanol aqueous solution as solvents, cinnamaldehyde was added dropwise to the solvents in a constant temperature water bath at 70 degrees Celsius to prepare 30 ml of different doses of drug solution.

[0062] A transparent cylindrical experimental tank with a diameter of 8.0 cm and a height of 9.5 cm was selected. The lid of the tank was perforated for ventilation (6 small holes with a diameter of 3.0 mm) as an experimental device to test the insecticidal activity of the combination of cinnamaldehyde and vermiculite against the adults of white-star flower beetles. 60 g of sterilized vermiculite was placed in the experimental tank as a matrix, and 30 ml of the drug solution at each dose was added respectively. The mixture was stirred evenly with a glass rod. After 30 min, the combination of cinnamaldehyde and vermiculite with different ratios was obtained. Six healthy adults of white-star flower beetles were placed in each treatment group. The survival of the adults of white-star flower beetles was recorded after 12 and 24 h. After gently picking up the adults, the adults were considered dead if their front tarsal claws were weak, they could not move normally, and their wings could not be unfolded normally. The experiment was repeated 5 times. Only 30 ml of solvent was added to 60 g of vermiculite as a negative control without cinnamaldehyde.

[0063] The results showed that the negative control (vermiculite) had no insecticidal effect on the adults of the white-spotted flower beetle. The adjusted mortality was calculated based on the negative control and the LC was calculated based on the adjusted mortality of cinnamaldehyde at each dose. 50 The results showed that in the cinnamaldehyde-vermiculite composite, the higher the cinnamaldehyde content, the higher the insecticidal toxicity to the adults of the white-star flower beetle, and the longer the time, the higher the insecticidal toxicity to the adults of the white-star flower beetle; the LC 50 is 18.50 mg / g (the unit is the mass ratio of cinnamaldehyde to the mass ratio of cinnamaldehyde and vermiculite), and the LC50 It is 2.73 mg / g (this unit is the ratio of the mass of cinnamaldehyde to the mass of the combination of cinnamaldehyde and vermiculite). Example 6: Selective tick control effect of cinnamaldehyde and vermiculite composition

[0064] Analyze and compare the LC of cinnamaldehyde in Example 4 against ticks and cinnamaldehyde in Example 5 against white-spotted flower beetle adults 50 Insecticidal activity, see Figure 4 The results showed that the presence of vermiculite can significantly enhance the insecticidal effect of cinnamaldehyde on ticks, and the sensitivity of adult white-spotted flower beetles to the combination of cinnamaldehyde and vermiculite is low, with a difference of more than 6 times. Therefore, the combination of cinnamaldehyde and vermiculite can be selected to selectively kill ticks without killing adult white-spotted flower beetles, achieving the effect of preventing and controlling ticks. Example 7: Control of ticks parasitic on adult white-spotted flower beetles

[0065] A transparent cylindrical experimental jar with a diameter of 8.0 cm and a height of 9.5 cm, and 6 ventilation holes with a diameter of 3.0 mm on the jar cover, was selected as an experimental device for testing the prevention and control of tick parasitism on adult white-spotted flower beetles.

[0066] After 22 adult ticks of the same age were placed in the experimental tank for 12 hours of starvation treatment, 1 adult white-spotted flower beetle was placed in the tank and placed in a standardized culture environment of 26±1 degrees Celsius, 50±5% humidity, and 16L:8D light cycle for 24 hours, and the parasitism rate of ticks on white-spotted flower beetles was counted. At this time, the parasitism rate of ticks on white-spotted flower beetles exceeded 50%, which was used as a standard parasitism model for further testing.

[0067] Using 1% Tween-80 and 10% anhydrous ethanol aqueous solution as solvents, cinnamaldehyde was added dropwise to the solvents in a constant temperature water bath at 70 degrees Celsius to prepare 10 ml of different doses of drug solution.

[0068] Take 20 g of sterilized vermiculite, add 10 ml of the solution at each dose to it, stir evenly with a glass rod, and place it for 30 minutes to obtain different ratios of cinnamaldehyde and vermiculite compositions. Fill the cinnamaldehyde and vermiculite compositions into the above-mentioned experimental jars containing tick-parasitized white-star flower beetle adults, and record the parasitism rate and mortality of ticks and the health status of white-star flower beetle adults after 12 and 24 hours of treatment. The experiment was repeated 12 times. Only 10 ml of solvent was added to 20 g of vermiculite as a negative control without cinnamaldehyde.

[0069] The results showed that after 12 hours of treatment, the parasitic rates of the cinnamaldehyde-vermiculite combination with cinnamaldehyde contents of 0.88 mg / g, 2.63 mg / g, and 4.38 mg / g decreased by 74.24%, 77.27%, and 81.82% respectively compared with the negative control group; after 24 hours of treatment, the parasitic rates of the three cinnamaldehyde contents decreased by 89.39%, 92.42%, and 95.45% respectively compared with the negative control group. In addition, the adults of the white-star flower beetle were in good health and had no obvious symptoms of poisoning when treated with the cinnamaldehyde-vermiculite combination with cinnamaldehyde contents of 0.88 mg / g and 2.63 mg / g; only one adult of the white-star flower beetle died in 24 hours when treated with the cinnamaldehyde-vermiculite combination with 4.38 mg / g.

[0070] The above data show that the combination of cinnamaldehyde and vermiculite can effectively and selectively prevent and control ticks. When a tick outbreak occurs, it is recommended to use 0.88 mg / g to 2.63 mg / g of the combination of cinnamaldehyde and vermiculite for 24 hours to prevent and control ticks without causing harm to the adult white-spotted flower beetles.

Claims

1. Application of cinnamaldehyde in the prevention and control of ticks.

2. The use according to claim 1, characterized in that: The tick is a species of Ixodes.

3. The use according to claim 2, characterized in that: The mitochondrial sequence of the tick is shown as SEQ ID No.

1.

4. A composition comprising cinnamaldehyde and vermiculite.

5. The composition according to claim 4, characterized in that Taking the mass of the composition as 100%, the content of cinnamaldehyde is greater than 0.4 mg / g.

6. The composition according to claim 4, characterized in that Taking the mass of the composition as 100%, the content of cinnamaldehyde is greater than 0.4 mg / g and less than 4 mg / g.

7. The composition according to claim 4, characterized in that Based on 100% by mass of the composition, the content of cinnamaldehyde is greater than 0.88 mg / g and less than 2.63 mg / g.

8. Use of the composition according to any one of claims 4 to 7 in controlling ticks.

9. The use according to claim 8, characterized in that: The tick is Ixodes.

10. The use according to claim 9, characterized in that The mitochondrial sequence of the tick is shown as SEQ ID No. 1.

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