Use of cinnamaldehyde and compositions containing same for the control of parasitic ticks

Abstract

The present application relates to the application of cinnamyl aldehyde and compositions containing the same in the prevention and treatment of parasitic ticks. The present application relates to the field of biocides. The present application first discovers that cinnamyl aldehyde has insecticidal activity on ticks, especially Ixodes, and is safe to the host of the ticks, white star flower beetle adults, at a certain dose, so that cinnamyl aldehyde can be used as an insecticide for ticks. Further, the present application also discovers that although vermiculite has no insecticidal activity on ticks, when cinnamyl aldehyde is added to vermiculite, the composition of cinnamyl aldehyde and vermiculite is used to prevent and treat ticks, and vermiculite can improve the insecticidal effect of cinnamyl aldehyde. Combined with the soil burrowing habit of white star flower beetle adults, vermiculite coincidentally provides a shelter for them, which provides operability for the prevention and treatment of parasitic ticks.

Description

Technical Field

[0001] This invention relates to the field of biocides, and particularly to the use of cinnamaldehyde and compositions containing it in the control of parasitic ticks. Background Technology

[0002] White-spotted Flower Beetle ( Protaetia brevitarsis *Protaetia spp.* (PB) is a soil-dwelling insect belonging to the class Insecta, order Coleoptera, family Cetoniidae, and genus *Protaetia*. Other names include White-striped Copper Beetle, White-spotted Flower Beetle, White-spotted Scarab Beetle, and Copper Beetle. It undergoes complete metamorphosis. Under natural conditions, adults typically infest sweet-smelling economic crops, while larvae thrive in environments with high organic matter content, such as decaying straw, fallen leaves, and livestock manure. The White-spotted Flower Beetle is widely distributed in Northeast China, North China, several eastern provinces along the southeast coast, and parts of Northwest China. It is also found in greater numbers in countries and regions such as Japan, Korea, Mongolia, and Russia.

[0003] The larvae of the white-spotted flower beetle are rich in nutrients, containing abundant protein, polypeptides, fatty acids, carbohydrates, alkaloids, antimicrobial peptides, vitamins, and trace amounts of minerals. Therefore, the larvae of the white-spotted flower beetle have a wide range of applications, playing a role in medicine, food, animal husbandry, agriculture and forestry, and environmental protection. Simultaneously, the larvae of the white-spotted flower beetle are a good source of protein. The protein content of the larvae is 57.86 ± 0.01% of their dry weight, similar to that of silkworms (55% to 76%) and locusts (68% to 78%), which are currently permitted for consumption. Furthermore, the protein content is similar to that of eggs, 1.80 times that of milk, and 2.34 times that of pork, making it an important source of novel protein resources, especially as animal feed. In 2018, the larvae of the white-spotted flower beetle were registered as a food ingredient by the Ministry of Food and Drug Safety (Korea). With further research, the agricultural value of the white-spotted flower beetle is being increasingly recognized, and its comprehensive application in agriculture is becoming more widespread.

[0004] The white star flower beetle larvae are commonly used medicinal materials, and are used in the preparation of Dahuangzhe pill, Ganqia pill, Shanzhu pill, and Tianluozi pill, etc. The white star flower beetle larvae are used in the prescription of Dahuangzhe pill, Dahuangzhe capsule, and Dahuangzhe tablet. In South Korea, the white star flower beetle larvae are a traditional medicinal material for Korean medicine. According to the records in Dongyibaojian, the white star flower beetle larvae can be used to treat liver cancer, liver cirrhosis, and hepatitis, relieve fatigue, and regulate menstrual disorders. The white star flower beetle larvae also have certain effects on visual impairment, cataract, postpartum wind, sore, stomatitis, and stroke, and have significant therapeutic or relieving effects on liver diseases, and are therefore commonly used in Korean medicine and folk prescriptions. In addition, the white star flower beetle larvae contain a large amount of health-related active substances, and the protein isolate of the white star flower beetle larvae can also be used as a potential antioxidant biomaterial for bone tissue engineering applications.

[0005] The white star flower beetle larvae can be developed into an organic fertilizer rich in nutrients, and can promote the development of the field of agricultural fertilizers.

[0006] In the breeding process of the white star flower beetle, the adult is easy to be parasitized by ticks, which can cause the death of the white star flower beetle population in severe cases, and is a big problem in the breeding of the white star flower beetle. These parasitic ticks are mainly parasitized in the foot base, thoracic and abdominal junction, and head and thoracic junction of the host, and suck the body fluid and blood lymph of the adult and the larvae to complete growth and reproduction, which seriously threatens the growth and development and health of the white star flower beetle. However, there is no product and technology for preventing and treating the parasitic ticks of the white star flower beetle at present, which restricts the development of the white star flower beetle industry. SUMMARY

[0007] The application provides an application of cinnamyl aldehyde in the prevention and treatment of ticks.

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

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

[0010] In one specific embodiment, the tick also has the following characteristics: the development life history thereof passes through four stages of egg, nymph tick, deutonymph tick, and adult tick; the egg is ellipsoidal or spherical, and is milky white or milk white, and the whole is translucent; the body of the newly hatched nymph tick is white and translucent, the nymph tick has three pairs of legs, the deutonymph tick and the adult tick have four pairs of legs; the length-width ratio of the body of the nymph tick and the deutonymph tick in the early stage is about 2:1, and the body is long and elliptical; the length-width ratio of the body of the deutonymph tick in the late stage and the adult tick is about 1.5:1, and the body is elliptical; the nymph tick and the deutonymph tick undergo two times of molting; the adult tick has an oval shape with a length of about 0.65 mm and a width of about 0.40 mm, and the body color is red-brown or gray-brown, and the mouth part is of a piercing-sucking type.

[0011] In one specific embodiment, the tick develops from egg to adult tick in about 40 to 45 days.

[0012] In one specific embodiment, the tick undergoes twice of ecdysis from larva to nymph and from nymph to adult.

[0013] In one specific embodiment, the tick has two parts of head and body, no division of thorax and abdomen, flat abdomen, and visible head from the back.

[0014] In one specific embodiment, the tick has strong movement ability, and the movement ability gradually increases as the tick develops to mature, the movement speed of larva is about 0.28 cm / s, and the movement speed of adult tick is about 0.46 cm / s.

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

[0016] In one specific embodiment, the content of cinnamaldehyde is more than 0.4 mg / g, based on 100% of the mass of the composition.

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

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

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

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

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

[0022] In one specific embodiment, the tick further has the following characteristics: its development life history goes through four stages of egg, larva, nymph and adult; the egg is ellipsoidal or spherical, milky white or cream white, and is translucent as a whole; the newly hatched larva is white and translucent, the larva has three pairs of legs, the nymph and adult have four pairs of legs; the body length-width ratio of the larva and early-stage nymph is about 2:1, and the shape is long elliptical; the body length-width ratio of the late-stage nymph and adult is about 1.5:1, and the shape is elliptical; the larva and nymph go through two molting processes, from larva to nymph and from nymph to adult; the adult tick is about 0.65 mm long and about 0.40 mm wide, is elliptical, has a body color of reddish brown or grayish brown, and has a piercing-sucking mouthpart.

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

[0024] In one specific embodiment, the tick goes through two molting processes, from larva to nymph and from nymph to adult.

[0025] In one specific embodiment, the tick has two parts of a false head and a body, and has no division of thorax and abdomen, and the abdomen is flat, and the false head can be seen from the back; the body is hard and smooth, and the outer edge of the tail end has about 12 body hairs of 50 microns long.

[0026] In one specific embodiment, the tick has a relatively strong movement ability as a whole, and the movement ability gradually increases as the body matures, and the movement speed of the larva is about 0.28 cm / s, and the movement speed of the adult is about 0.46 cm / s.

[0027] Advantages of the present application: the present application first discovers that cinnamyl aldehyde has insecticidal activity on ticks, especially Ixodes, and is safe to the host Anomala daimiana Motschulsky adult at a certain dose, so that cinnamyl aldehyde can be used as a tick insecticide. However, when using contact killing method to kill ticks parasitizing on the body of Anomala daimiana Motschulsky adult, since the parasitization of ticks usually occurs at sites such as the foot base, thorax-abdomen joint and head-thorax joint of Anomala daimiana Motschulsky adult, it is difficult to reach these parasitized sites of Anomala daimiana Motschulsky adult by spraying method, so it is difficult to achieve a killing effect comparable to that under non-parasitization conditions in actual operation; fumigation for killing ticks needs to ensure the airtightness of the space for killing ticks, otherwise the concentration of cinnamyl aldehyde in the space cannot be guaranteed, but the oxygen supply to Anomala daimiana Motschulsky adult cannot be guaranteed in such an airtight space, which affects the health of Anomala daimiana Motschulsky adult. Further, the present application also finds that although vermiculite has no insecticidal activity on ticks, when cinnamyl aldehyde is added to vermiculite, and the combination of cinnamyl aldehyde and vermiculite is used to control ticks, the vermiculite can improve the insecticidal effect of cinnamyl aldehyde. Combined with the soil-pot habit of Anomala daimiana Motschulsky adult, the vermiculite coincidentally provides a sheltering place for Anomala daimiana Motschulsky adult, and provides operability for controlling parasitic ticks. BRIEF DESCRIPTION OF DRAWINGS

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

[0029] Figure 2 The parasitic rate of ticks on WSFC adults is shown.

[0030] Figure 3 The results of the alignment of the mitochondrial sequence of the test tick of the present application with the mitochondrial sequences of known tick species are shown.

[0031] Figure 4 The LC50 of cinnamyl aldehyde on ticks in Example 4 and on WSFC adults in Example 5 are shown. 50 Comparison of insecticidal activity. DETAILED DESCRIPTION

[0032] The above content of the present application is further described in detail in the form of preferred embodiments, but it does not constitute a limitation on the present application.

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

[0034] Test insects: WSFC adults were collected from the insect rearing room of the Langfang Pilot Base of the Chinese Academy of Agricultural Sciences, and after the eggs were hatched, the larvae were grown to the third instar pupae, and then the adults were obtained after eclosion.

[0035] Test ticks: The parasitic ticks of the present application were isolated from WSFC adults reared at the Langfang Pilot Base of the Chinese Academy of Agricultural Sciences and were reared in vitro.

[0036] Tick feed: Chicken and poultry bone skeletons and bone marrow were crushed into powder, and an equal amount of pure water was added to prepare chicken and poultry bone homogenate. Taking the total mass of the chicken and poultry bone homogenate as 100%, 0.1wt% potassium sorbate was added as a preservative, 1.0wt% jelly powder, 121℃ sterilization treatment for 15 min, poured into a mold, cooled, solidified, shaped, and stored in a 4℃ refrigerator.

[0037] Test tick rearing: Natural fermented edible fungus residues with a humidity of 50% were used as rearing substrate, 200 g of rearing substrate was taken, and an appropriate amount of tick feed was added, and ticks were inoculated into the tick feed from the body surface of WSFC adults parasitized by ticks, and placed in an artificial climate chamber with a temperature of 26±1℃, a humidity of 50±5%, and a light cycle of 16L:8D for cultivation. According to the experimental requirements, the ticks were collected from the substrate, and the eggs, nymphs, and adults were tested.

[0038] Tick inoculation to white star flower beetle adults: healthy white star flower beetle adults were put into the breeding substrate with ticks for 24 hours, and the white star flower beetle adults were inoculated with ticks for further testing.

[0039] Cinnamaldehyde, density 1.05 g / ml, purchased from Shengong Bioengineering (Shanghai) Co., Ltd.

[0040] Magnetic Universal Genomic DNA Kit for extracting genomic DNA was purchased from TIANGEN.

[0041] Rapid Plus DNA Lib Prep Kit for Illumina (RK20208) for tick DNA sequencing was purchased from Abclonal.

[0042] Example 1: Identification of the parasitic tick population of the white star flower beetle

[0043] 1wt% agar plate was poured into a 35 mm culture dish as a substrate, and after the agar plate was solidified, 3.0 g of tick feed was placed in the center, and 30 adult ticks isolated from white star flower beetle adults and artificially cultured were introduced, and the feeding conditions were environmental temperature 26±1 degrees Celsius, humidity 50±5%, and photoperiod 16L:8D, for observing the developmental life history of the parasitic tick.

[0044] The biological characteristics of the parasitic tick involved in the present application: the developmental life history of the parasitic tick was determined by artificial culture observation to pass through four stages of egg, larva, nymph, and adult. Among them, it takes about 40 to 45 days from egg to adult. The egg is ellipsoidal or spherical, milky white or cream white, and the whole is translucent. The newly hatched larva is white and translucent, the larva has three pairs of legs, the nymph has four pairs of legs, and the adult has four pairs of legs. The length-width ratio of the larva and the early nymph is about 2:1, and the shape is long oval, and the length-width ratio of the late nymph and the adult is about 1.5:1, and the shape is oval. Two times of molting from larva to nymph and from nymph to adult, the body color is cream white within a few days after molting. The adult tick is about 0.65 mm long and about 0.40 mm wide, oval, body color red-brown or gray-brown, and piercing-sucking mouthparts. The whole tick is divided into two parts of false head and body, without head, thorax and abdomen, the ventral surface is flat, and the false head can be seen from the dorsal view. The body is hard and smooth, and there are about 12 body hairs of 50 microns long on the dorsal side of the outer edge of the tail end. The overall movement ability of the tick is relatively strong, and gradually increases with the development and maturation of the insect body, the moving speed of the larva is about 0.28 cm / s, and the moving speed of the adult is about 0.46 cm / s. Figure 1 Each stage of the tick life history is shown.

[0045] Observations show that, in their natural state, the larvae, nymphs, and adults of the parasitic ticks involved in this invention feed on the host, and their bodies swell slightly after feeding. Ticks are photophobic, gregarious, and prefer damp environments; they curl up and feign death when exposed to external physical stimuli. Parasitic ticks lay their eggs in shady places (such as under stones, clods of earth, or in wall crevices). After mating, the female tick falls to the ground to lay eggs after feeding.

[0046] Verification of the parasitic ability of ticks: Healthy individuals of the white-spotted flower beetle were obtained from the indoor breeding room of the white-spotted flower beetle at the Langfang Pilot Base of the Chinese Academy of Agricultural Sciences. Two 35 mm petri dishes were placed mouth-to-mouth, one with water agar bottom for easy tick inoculation. The two dishes were connected at the junction with breathable tape to serve as the tick rearing device. Twenty ticks of the same age were introduced into the rearing device and starved for 12 hours. Then, one adult WSFC tick was introduced. The rearing device was placed in a standardized culture environment of 26±1°C, 50±5% humidity, and 16L:8D photoperiod to serve as a standard parasitic model. The tick parasitic behavior was observed and recorded at 3, 6, 12, 24, 48, and 72 hours after the start of the experiment, and the parasitism rate was calculated. The experiment was repeated 7 times. Parasitism rate (%) = 100 × number of host ticks / total number of ticks. The results are shown in the table below. Figure 2 .

[0047] Observations revealed that tick parasitism mainly occurred at the base of the feet, the junction of the thorax and abdomen, and the junction of the head and thorax of the white-spotted flower beetle, similar to the situation observed during breeding, indicating that the ticks isolated in this invention are the main harmful species.

[0048] Figure 2 The results showed that the parasitism rates of larvae, nymphs, and adults all increased with increasing treatment time. Within 6 hours of treatment, the parasitism rates of the three species were relatively low, at 17.86%, 29.29%, and 23.57%, respectively, and there was no significant difference compared with the parasitism rates after 3 hours of treatment. P >0.05). The parasitism rate gradually stabilized after 48 hours of treatment, with rates of 92.14%, 87.86%, and 85.71% at 72 hours, respectively. Among the treatments for different tick infestation ages, the parasitism rate showed the most significant difference at 12 hours, with larvae having the lowest parasitism rate at 25.00% and juvenile ticks having the highest at 63.29%. The parasitism rate of larvae increased most significantly during 12 and 24 hours of treatment, with a highly significant difference between the two periods. P <0.01). It is evident that all developmental stages of the parasitic tick exhibit parasitic behavior against the white-spotted flower beetle. Among them, larvae and nymphs are more active in parasitism than adults, with nymphs causing the fastest damage. Under long-term treatment, larvae exhibit the highest parasitism rate. From the perspective of infection speed, nymphs have the fastest infection rate, exceeding 50% after only 12 hours of treatment; however, because adult ticks are relatively larger and easier to handle, subsequent experiments used adult ticks as the experimental subjects.

[0049] White star flower beetle parasitic mite species analysis: the whole genome of the parasitic mite cultured under standardized in vitro conditions was extracted, high-throughput sequencing was performed, the sample library was constructed using Rapid Plus DNA Lib Prep Kit for Illumina (RK20208) kit, the mitochondria was assembled and annotated using Mitoz software, and a complete 14438 bp circular mitochondrial sequence (as shown in SEQ ID No. 1) was obtained, which has 13 CDS, 2 rRNA (I-rRNA and s-rRNA), and 22 tRNA. Sequence analysis was performed using the BLAST online platform of the National Center for Biotechnology Information, and the results are shown in Figure 3 . Figure 3 The mitochondrial sequence alignment results of Ixodes rubicundus show that the test mite has the highest overall score compared with Ixodes , but the sequence alignment similarity with Ixodes is lower than 80%, so the test mite is a new species of Ixodes.

[0050] Example 2: Determination of contact toxicity of the agent

[0051] Potter spray method was used for contact toxicity determination, and the mortality of mites at 12 h and 24 h was recorded, and the contact LC 50 .

[0052] Using 1% Tween-80, 10% anhydrous ethanol aqueous solution as solvent, 70 degrees Celsius constant temperature water bath environment, adding cinnamyl aldehyde to the solvent to prepare seven different gradient concentrations of drug solution.

[0053] Ten adult mites were directly inserted into a 35 mm diameter parasitic mite feeding device, and sprayed under a Potter spray tower (pressure 100 kPa, each spray volume 100 microliters, settlement 2 min), and the experiment was repeated 7 times. The solvent treatment was used as a negative control, and then placed in a 26 degrees Celsius constant temperature incubator for feeding, and the results were investigated after 12 and 24 h, respectively, the number of dead mites was recorded, the corrected mortality was calculated, and the LC 50 was calculated based on the corrected mortality at each concentration, and the experiment was repeated 5 times. Among them, those who lost the reaction after touching with a hairbrush, could not normally crawl, or had body surface shrinkage, discoloration, deformation, and internal contents excretion were judged as dead. Mortality (%) = number of dead insects / total number of insects x 100; corrected mortality (%) = 100 x (treatment group mortality-control group mortality) / (1-control group mortality).

[0054] The results show that under the spray treatment, cinnamyl aldehyde has significant contact toxicity against the test mite, and the contact toxicity increases with time. The LC 50LC at 24 h treatment was 5.50 g / L 50 LC at 24 h treatment was 4.27 g / L.

[0055] Example 3: Fumigant activity determination of the agent

[0056] The fumigant activity determination was carried out by using the Petri dish closed fumigation method, and the mortality of ticks at 12 h and 24 h was recorded, and the fumigant LC 50 was calculated.

[0057] A piece of filter paper (length 1.0 cm x width 0.5 cm) was sterilized and fixed on the parasitic tick rearing device with a diameter of 35 mm by double-sided tape, and the filter paper strip was suspended in the space of the Petri dish to facilitate the volatilization of cinnamyl aldehyde. Among them, the concentration of cinnamyl aldehyde was calculated based on the mass of cinnamyl aldehyde added to the filter paper strip and the volume of the space of the Petri dish. 10 ticks were introduced into the Petri dish, and then different amounts of cinnamyl aldehyde were added to the middle of the filter paper strip, and the Petri dish was quickly sealed with sealing film and placed in a 26°C constant temperature incubator for rearing. The results were investigated at 12 and 24 h, respectively, the number of dead ticks was recorded, the corrected mortality was calculated, and the LC 50 was calculated based on the corrected mortality at each concentration. The experiment was repeated 5 times.

[0058] The results showed that under the fumigation treatment, cinnamyl aldehyde had significant fumigant toxicity to the test ticks, and the toxicity increased with time. The LC 50 at 12 h treatment was 320.6 mg / L, and the LC 50 at 24 h treatment was 238.5 mg / L.

[0059] Example 4: Activity determination of cinnamyl aldehyde and vermiculite composition on ticks

[0060] The toxicity of cinnamyl aldehyde and vermiculite composition to ticks was tested. The mortality of ticks at 12 and 24 h was recorded respectively, and the LC 50 to ticks was calculated.

[0061] Using 1% Tween-80, 10% anhydrous ethanol aqueous solution as solvent, 70°C constant temperature water bath environment, cinnamyl aldehyde was added dropwise into the solvent to prepare 10 ml of different doses of drug solution.

[0062] Preparation 20 g sterilized vermiculite as a matrix, respectively, add the drug solution 10 ml, stirring with a glass rod, placed 30 min to make the vermiculite fully absorb the free drug solution, get different ratio of cinnamyl aldehyde and vermiculite composition. 1.0 g each dose of cinnamyl aldehyde and vermiculite composition were filled into the parasitic tick feeding device, and 10 ticks were introduced into it, and the number of dead ticks was recorded after 12 and 24 h, respectively, and the experiment was repeated 5 times. Among them, only 10 ml of solvent was added to 20 g of vermiculite as a negative control without cinnamyl aldehyde.

[0063] The results showed that the negative control (vermiculite) had no insecticidal effect on ticks. The corrected mortality rate was calculated based on the negative control and the LC 50 was calculated based on the corrected mortality rate of cinnamyl aldehyde at each dose. 50 The results showed that in the cinnamyl aldehyde and vermiculite composition, the higher the content of cinnamyl aldehyde, the higher the insecticidal toxicity to the test ticks; the LC 50 was 2.76 mg / g (the unit was the mass ratio of cinnamyl aldehyde to the mass ratio of cinnamyl aldehyde and vermiculite composition) after 12 h treatment, and the LC 50 was 0.44 mg / g (the unit was the mass ratio of cinnamyl aldehyde to the mass ratio of cinnamyl aldehyde and vermiculite composition) after 24 h treatment.

[0064] Example 5: Activity determination of cinnamyl aldehyde and vermiculite composition on white star flower beetle adults

[0065] The toxicity of cinnamyl aldehyde and vermiculite composition to healthy white star flower beetle adults was tested, and the mortality rate of white star flower beetle adults was recorded after 12 h and 24 h, and the LC 50 of white star flower beetle adults was calculated.

[0066] Using 1% Tween-80, 10% anhydrous ethanol aqueous solution as solvent, 70 degrees Celsius constant temperature water bath environment, dropwise adding cinnamyl aldehyde into the solvent to prepare 30 ml different doses of drug solution.

[0067] Select a transparent column body experimental tank with a diameter of 8.0 cm and a height of 9.5 cm, the tank cover is perforated for ventilation (6 small holes with a diameter of 3.0 mm), as the experimental device for testing the insecticidal activity of cinnamyl aldehyde and vermiculite composition. 60 g sterilized vermiculite was placed in the experimental tank as a matrix, and 30 ml of drug solution at each dose was added, respectively, and stirred uniformly with a glass rod, and placed for 30 min to obtain different ratios of cinnamyl aldehyde and vermiculite composition. 6 healthy white star flower beetle adults were placed in each treatment group, and the survival of white star flower beetle adults was recorded after 12 and 24 h, respectively. The adults were gently picked up, and the adults were considered dead if the front tarsal claw was weak and could not move normally, and the wings could not be normally unfolded. The experiment was repeated 5 times. Only 30 ml of solvent was added to 60 g of vermiculite as a negative control without cinnamyl aldehyde.

[0068] The results show that the negative control (vermiculite) has no insecticidal effect on the adult of Protaetia brevitabris. The corrected mortality rate is calculated based on the negative control, and the LC 50 is calculated based on the corrected mortality rate of cinnamyl aldehyde at each dose. The results show that in the cinnamyl aldehyde and vermiculite composition, the higher the content of cinnamyl aldehyde, the higher the insecticidal toxicity to the adult of Protaetia brevitabris, and the longer the time, the higher the insecticidal toxicity to the adult of Protaetia brevitabris; the LC 50 is 18.50 mg / g (the unit is the mass ratio of cinnamyl aldehyde to the mass of the cinnamyl aldehyde and vermiculite composition) at 12 h, and the LC 50 is 2.73 mg / g (the unit is the mass ratio of cinnamyl aldehyde to the mass of the cinnamyl aldehyde and vermiculite composition) at 24 h.

[0069] Example 6: Selective killing effect of cinnamyl aldehyde and vermiculite composition on ticks

[0070] The LC 50 insecticidal activity of cinnamyl aldehyde on ticks in the above-mentioned Example 4 and the LC Figure 4 insecticidal activity of cinnamyl aldehyde on the adult of Protaetia brevitabris in Example 5 are analyzed and compared. The results show that the presence of vermiculite can significantly improve the insecticidal effect of cinnamyl aldehyde on ticks, and the adult of Protaetia brevitabris has a lower sensitivity to the composition of cinnamyl aldehyde and vermiculite, with a difference of more than 6 times. Therefore, the composition of cinnamyl aldehyde and vermiculite can be selected to selectively kill ticks without killing the adult of Protaetia brevitabris, so as to achieve the effect of controlling ticks.

[0071] Example 7: Controlling ticks parasitizing the adult of Protaetia brevitabris

[0072] A transparent column experimental tank with a diameter of 8.0 cm and a height of 9.5 cm is selected as the experimental device for testing the control of ticks parasitizing the adult of Protaetia brevitabris, and 6 air holes with a diameter of 3.0 mm are punched on the tank cover.

[0073] After 12 h of starvation treatment of the same age of adult ticks 22 per tank, one adult of Protaetia brevitabris is put into each tank, and the tank is placed in a standardized culture environment with a temperature of 26±1 degrees Celsius, a humidity of 50±5%, and a light cycle of 16L:8D for 24 h. The parasitization rate of ticks parasitizing the adult of Protaetia brevitabris is counted. At this time, the parasitization rate of ticks parasitizing the adult of Protaetia brevitabris is more than 50%, which is used as a standard parasitization model for further testing.

[0074] 1% Tween-80, 10% anhydrous ethanol aqueous solution is used as a solvent, and cinnamyl aldehyde is added dropwise into the solvent to prepare 10 ml of different doses of drug solution in a constant temperature water bath environment at 70 degrees Celsius.

[0075] Take 20 g of sterilized vermiculite, add 10 ml of each dose of the drug solution to it, stir it evenly with a glass rod, and place it for 30 min to obtain different ratios of cinnamyl aldehyde and vermiculite compositions. Fill the cinnamyl aldehyde and vermiculite compositions into the experimental jars containing the white star flower beetle adults parasitized by ticks, respectively, and record the parasitic rate and mortality of ticks after 12 and 24 h of treatment. The experiment was repeated 12 times. Add 10 ml of solvent to 20 g of vermiculite as a negative control without cinnamyl aldehyde.

[0076] The results show that after 12 h of treatment, the parasitic rate of cinnamyl aldehyde and vermiculite compositions with cinnamyl aldehyde content of 0.88 mg / g, 2.63 mg / g, and 4.38 mg / g decreased by 74.24%, 77.27%, and 81.82% compared with the negative control group, respectively. After 24 h of treatment, the parasitic rate of the three cinnamyl aldehyde content groups decreased by 89.39%, 92.42%, and 95.45% compared with the negative control group, respectively. Moreover, the white star flower beetle adults were in good health without obvious poisoning symptoms under the treatment of cinnamyl aldehyde and vermiculite compositions with cinnamyl aldehyde content of 0.88 mg / g and 2.63 mg / g. Only one white star flower beetle adult died after 24 h of treatment with cinnamyl aldehyde and vermiculite compositions with cinnamyl aldehyde content of 4.38 mg / g.

[0077] The above data show that cinnamyl aldehyde and vermiculite compositions can effectively and selectively control ticks. When the tick outbreak occurs, it is recommended to use 0.88 mg / g to 2.63 mg / g of cinnamyl aldehyde and vermiculite compositions for 24 h of treatment to achieve the purpose of controlling ticks and not harming white star flower beetle adults.

Claims

1. The use of a composition for controlling ticks parasitizing adult white-spotted beetles, said composition comprising cinnamaldehyde and vermiculite, wherein the cinnamaldehyde content is above 0.4 mg / g and below 4 mg / g, based on the mass of said composition.

2. The application according to claim 1, characterized in that, The content of cinnamaldehyde, based on the mass of the composition, is 100% above 0.88 mg / g and below 2.63 mg / g.

3. The application according to claim 1, characterized in that, The ticks mentioned are hard ticks (Ixodes).

4. The application according to claim 3, characterized in that, The mitochondrial sequence of the tick is shown in SEQ ID No. 1.

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