Pharmaceutical composition and pharmaceutical preparation for killing chicken skin thorn mites and preparation method of pharmaceutical composition and pharmaceutical preparation

Abstract

The invention relates to the technical field of biopesticide materials, in particular to a pharmaceutical composition and a pharmaceutical preparation for killing chicken skin thorn mites and a preparation method of the pharmaceutical composition and the pharmaceutical preparation. The pharmaceutical composition for killing the chicken skin thorn mites comprises at least two of cinnamon essential oil, origanum essential oil or carbamate compounds. The pharmaceutical preparation prepared from the pharmaceutical composition for killing the chicken skin thorn mites is good in mite killing effect, free of toxic and harmful effects on the environment and non-target animals and capable of achieving the long-acting mite killing effect. According to the invention, the problems of low mite killing rate, environmental unfriendliness and toxic threat to animals in the chemical method for killing the chicken skin thorn mites in the prior art are effectively solved, and a new thought is provided for a novel environment-friendly acaricide.

Description

Technical Field

[0001] The invention relates to the technical field of biological pesticide materials, and in particular to a pharmaceutical composition and a pharmaceutical preparation for killing Dermanyssus gallinae and a preparation method thereof. Background Art

[0002] Dermanyssus gallinae, also known as poultry red mite, is an external parasite of chickens. It has a short life cycle, reproduces rapidly, and is very harmful to chickens. It sucks blood and its bites cause itching, which leads to the chickens pecking themselves and each other, reducing their growth rate or egg production rate, and easily causing huge economic losses to laying hen farming.

[0003] Within the existing technical framework, chemical acaricides are widely used to prevent and control Dermatophytes gallinae. However, long-term reliance on such drugs has caused multiple problems: the efficiency of killing acarids has gradually declined, while the degree of harm to non-target animals and environmental pollution has become increasingly serious. In particular, pyrethroid drugs, typified by deltamethrin, although showing strong insecticidal efficacy and relatively low toxicity, pose a fatal threat to fish and are irritating to human skin, mucous membranes and respiratory tract. Their residues may also lurk in eggs, causing food safety concerns. In addition, there are macrolide drugs such as avermectin and ivermectin, which, if used carelessly, can easily induce stress reactions and even poisoning incidents in chickens. Given their high toxicity and difficulty in natural degradation, the residues of such drugs in water bodies will profoundly affect the balance of aquatic ecosystems, and may be transmitted and accumulated through the food chain, posing a long-term potential hazard to the health of humans and other organisms.

[0004] Due to the above factors, the development of a safer, more environmentally friendly and highly effective mite-killing drug preparation is of great practical significance for the prevention and control of Dermanyssus gallinae. Summary of the invention

[0005] In view of the problems of low killing rate, environmental unfriendliness and toxicity threat to animals in the chemical method for killing Dermanyssus gallinae in the prior art, the present invention provides a pharmaceutical composition, a pharmaceutical preparation and a preparation method thereof for killing Dermanyssus gallinae. The pharmaceutical composition is prepared by compounding cinnamon essential oil, oregano essential oil or carbamate compounds as raw materials. Cinnamon essential oil and oregano essential oil are both natural plant essential oils, which are easy to degrade, have good killing effect on Dermanyssus gallinae, and are relatively safe for humans and animals. Carbamate compounds have the characteristics of low toxicity, low residue and long residual effect. The pharmaceutical preparation prepared by using the pharmaceutical composition for killing Dermanyssus gallinae not only has good killing effect, is friendly to the environment and animals, and can achieve high-efficiency killing of mites, providing a new idea for new environmentally friendly miticides.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] A first aspect of the present invention provides a pharmaceutical composition for killing Dermanyssus gallinae, wherein the pharmaceutical composition comprises at least two of cinnamon essential oil, oregano essential oil or carbamate compounds.

[0008] Plant essential oils are plant secondary metabolites that contain a variety of terpene compounds with strong surface activity and fat solubility, which enables them to quickly penetrate and destroy the skin cell membrane of Dermatosus gallinae. Once penetrated into the cells, these active ingredients will destroy the structure of the cell membrane, leading to the loss of cell contents, thereby destroying the physiological functions of the mites. In addition, these active ingredients can inhibit the activity of acetylcholinesterase in the nervous system of Dermatosus gallinae, thereby achieving a good killing effect of Dermatosus gallinae. Plant essential oils are environmentally friendly, low in toxicity, easily degradable, and relatively safe for humans and animals, avoiding the problem of traditional acaricides that are highly toxic, have long-lasting residues, and are potentially harmful to health after contact or inhalation by humans or animals.

[0009] Carbamate compounds have the characteristics of broad-spectrum and high-efficiency insecticide, low toxicity, low residue and long residual effect. After entering the body of Dermatosus gallinae, they inhibit the cholinesterase of the nervous system of Dermatosus gallinae and cause its death. The present invention prepares the pharmaceutical composition for killing Dermatosus gallinae by compounding cinnamon essential oil, oregano essential oil or carbamate compounds. The pharmaceutical composition has the effect of killing Dermatosus gallinae efficiently and is friendly to the environment and animals, providing a new idea for a new environmentally friendly acaricide.

[0010] Preferably, the pharmaceutical composition consists of cinnamon essential oil and oregano essential oil.

[0011] Preferably, the pharmaceutical composition consists of cinnamon essential oil and carbamate compounds.

[0012] Preferably, the pharmaceutical composition consists of oregano essential oil and carbamate compounds.

[0013] Preferably, the carbamate compound is (1-naphthyl)-N-methylcarbamate.

[0014] The second aspect of the present invention provides a pharmaceutical preparation for killing Dermanyssus gallinae, wherein the pharmaceutical preparation comprises the pharmaceutical composition for killing Dermanyssus gallinae.

[0015] Preferably, the pharmaceutical preparation further comprises a diluent.

[0016] Preferably, when the pharmaceutical composition consists of cinnamon essential oil and oregano essential oil, the final concentration of cinnamon essential oil in the pharmaceutical preparation is 14 mg / mL-22 mg / mL, and the final concentration of oregano essential oil is 2 mg / mL-12 mg / mL.

[0017] Further preferably, when the pharmaceutical composition consists of cinnamon essential oil and oregano essential oil, the final concentration of cinnamon essential oil in the pharmaceutical preparation is 16.8 mg / mL, and the final concentration of oregano essential oil is 8.4 mg / mL.

[0018] Preferably, when the pharmaceutical composition consists of cinnamon essential oil and carbamate compounds, the final concentration of cinnamon essential oil in the pharmaceutical preparation is 2 mg / mL-8 mg / mL, and the final concentration of carbamate compounds is 7 μg / mL-9 μg / mL.

[0019] Further preferably, when the pharmaceutical composition consists of cinnamon essential oil and carbamate compounds, the final concentration of cinnamon essential oil in the pharmaceutical preparation is 4.8 mg / mL, and the final concentration of carbamate compounds is 8 μg / mL.

[0020] Preferably, when the pharmaceutical composition comprises oregano essential oil and carbamate compounds, the final concentration of oregano essential oil in the pharmaceutical preparation is 14 mg / mL-26 mg / mL, and the final concentration of carbamate compounds is 1 μg / mL-5 μg / mL.

[0021] Further preferably, when the pharmaceutical composition comprises oregano essential oil and carbamate compounds, the final concentration of oregano essential oil in the pharmaceutical preparation is 16.8 mg / mL, and the final concentration of carbamate compounds is 4 μg / mL.

[0022] The pharmaceutical preparation for killing Dermanysus gallinae provided by the present invention uses at least two of cinnamon essential oil, oregano essential oil or carbamate compounds as raw materials, and the median lethal concentration (LC50) of the three agents on Dermanysus gallinae is measured. 50 ), based on which, the pharmaceutical preparation is prepared by adjusting the volume ratio of the two components. The pharmaceutical preparation can achieve high efficiency in killing mites, and can also alleviate the drug resistance problem caused by lack of drugs while reducing costs.

[0023] Preferably, the diluent is acetone.

[0024] The third aspect of the present invention provides a method for preparing the pharmaceutical preparation for killing Dermanyssus gallinae, comprising the following steps:

[0025] S1, dissolving cinnamon essential oil, oregano essential oil and carbamate compound in diluent respectively to obtain cinnamon essential oil solution, oregano essential oil solution and carbamate compound solution;

[0026] S2. Mix at least two of the cinnamon essential oil solution, the oregano essential oil solution or the carbamate compound solution to obtain a pharmaceutical preparation for killing Dermanyssus gallinae.

[0027] In summary, the present invention provides a pharmaceutical composition for killing Dermanyssus gallinae, a pharmaceutical preparation and a preparation method thereof, wherein the pharmaceutical composition for killing Dermanyssus gallinae is prepared by compounding plant essential oils and carbamate compounds as raw materials. The pharmaceutical preparation prepared by using the pharmaceutical composition for killing Dermanyssus gallinae not only has a good mite killing effect, is friendly to the environment and animals, and can achieve long-term mite killing. The technical solution provided by the present invention effectively solves the problems of low mite killing rate, unfriendly environment and toxicity threat to animals in the chemical method for removing Dermanyssus gallinae in the prior art, and provides a new idea for the prevention and control of Dermanyssus gallinae. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 A graph showing the mortality of Dermanyssus gallinae treated with the composite agent provided in Examples 1 to 9;

[0029] Figure 2 A graph showing the mortality of Dermanyssus gallinae treated with the composite agent provided in Examples 10 to 18;

[0030] Figure 3 A synergistic effect diagram of the composite agent provided in Examples 1 to 9 in treating Dermanyssus gallinae;

[0031] Figure 4 A synergistic effect diagram of the composite agent provided in Examples 10 to 18 in treating Dermanyssus gallinae;

[0032] Figure 5 Mortality graph of Dermanyssus gallinae treated with the composite agent provided in Examples 19 to 27

[0033] Figure 6 A synergistic effect diagram of the composite agent provided in Examples 19 to 27 in treating Dermanyssus gallinae;

[0034] Figure 7 This is a graph showing the fumigation toxicity test of the composite agents provided in Examples 1 to 9 on Dermanyssus gallinae at different time periods;

[0035] Figure 8 This is a graph showing the fumigation toxicity test of the composite agents provided in Examples 10 to 18 on Dermanyssus gallinae at different time periods;

[0036] Fig. 9 This is a graph showing the fumigation toxicity test of the composite agents provided in Examples 19 to 27 on Dermanyssus gallinae at different time periods;

[0037] Fig.10The figures are the hatching results of the eggs obtained after being treated with the composite agents provided in Example 2 (E1), Example 15 (E2), Example 25 (E3), Comparative Example 1 (A1), Comparative Example 2 (A2) and Comparative Example 3 (A3), and the hatching results of the eggs obtained after being treated with the acetone blank control group (C1), the ethanol blank control group (C2) and the distilled water blank control group (C3);

[0038] Fig.11 The schematic diagram of the morphology of insect eggs after being treated with the composite agents provided in Example 2, Example 15, Example 25, Comparative Example 1, Comparative Example 2 and Comparative Example 3 for two days, and the morphology of insect eggs after being treated with the acetone blank control group, the ethanol blank control group and the distilled water blank control group for two days; Fig.11 (A) is the morphology of eggs after two days of treatment with the composite agent provided in Example 2; Fig.11 (B) is the morphology of eggs after two days of treatment with the composite agent provided in Example 15; Fig.11 (C) The morphology of eggs two days after treatment with the compound agent provided in 25; Fig.11 (D) is the morphology of eggs after two days of treatment with the agent provided in Comparative Example 1; Fig.11 (E) is the morphology of eggs after two days of treatment with the agent provided in Comparative Example 2; Fig.11 (F) is the morphology of eggs after two days of treatment with the agent provided in Comparative Example 3; Fig.11 (G) shows the morphology of eggs after two days of acetone treatment; Fig.11 (H) shows the morphology of eggs after two days of ethanol treatment; Fig.11 (I) shows the morphology of eggs after two days of treatment with distilled water. DETAILED DESCRIPTION

[0039] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0040] 1.1 Contact toxicity of three single agents to Dermanyssus gallinae

[0041] The filter paper contact method was used to determine the toxic effects of three single doses on Dermanyssus gallinae. Two circular filter papers with a diameter of 37 mm were used to make a biological toxicity determination device with a "hat" with a circular depression with a central diameter of 25 mm and a lower "lid". 5-7 gradient concentrations of essential oil solutions and drug solutions were prepared, and 100 μL of the same gradient concentration drug solution was evenly applied to two filter papers. After drying, the two were glued together. The top of the upper "hat" was opened and used as the entrance for mites. 15 adult female mites were picked and the opening was sealed with transparent tape. The treatment was carried out for 24 hours. The number of dead chicken mites was recorded. Death was considered if the limbs were curled up or any part was inactive; and survival was considered if any part of the body was active (trembling, crawling). SPSS software was used to calculate the LC values ​​of the three single doses on Dermanyssus gallinae. 50 .

[0042] The results are shown in Table 1, where LC 50 and LC 90 : concentration required to achieve 50% and 90% mortality of mites; CI: 95% confidence interval; χ 2 (df): chi-square value and degrees of freedom; R 2 : regression coefficient. Table 1 shows the lethal effects of three single-dose drugs directly acting on Dermanyssus gallinae. The LC values ​​of each single-dose drug are 50 They are: cinnamon essential oil concentration is 23.9 mg / mL; oregano essential oil concentration is 27.6 mg / mL; (naphthyl)-N-methylcarbamic acid concentration is 8.2 μg / mL.

[0043] Table 1 Results of contact toxicity test of single-dose drugs on Dermanyssus gallinae

[0044]

[0045] The specific ingredients of the pharmaceutical preparation for killing Dermanyssus gallinae provided by the present invention are shown in Table 2.

[0046] Table 2 Statistics of ingredients in various embodiments and comparative examples

[0047]

[0048]

[0049] Wherein, the cinnamon essential oil solution is cinnamon essential oil LC 50 The solution comprises the following steps: dissolving 23.9 g of pure cinnamon essential oil in 1 L of acetone to obtain a cinnamon essential oil solution with a concentration of 23.9 g / L.

[0050] The oregano essential oil solution is oregano essential oil LC 50 The solution comprises the following steps: dissolving 27.6 g of pure oregano essential oil in 1 L of acetone to obtain an oregano essential oil solution with a concentration of 27.6 g / L.

[0051] The (1-naphthyl)-N-methylcarbamate solution is (1-naphthyl)-N-methylcarbamate LC 50 The solution comprises the following steps: dissolving 8.2 mg of (1-naphthyl)-N-methylcarbamate in 1 L of acetone to obtain a (1-naphthyl)-N-methylcarbamate solution with a concentration of 8.2 mg / L.

[0052] The present invention conducted the following tests on the pharmaceutical preparations obtained in each embodiment and comparative example:

[0053] 1.2 Example Drug Contact Toxicity

[0054] The present invention conducted a contact toxicity test on the pharmaceutical preparations provided in the examples and comparative examples, and the test method was the contact toxicity test method described in 1.1. The combined effect between the combinations was evaluated by calculating the toxicity-effect ratio. The calculation was performed according to the following equation:

[0055] Theoretical mortality rate of mixture M = mortality rate of mites after treatment with solution A × proportion of A in M ​​+ mortality rate of mites after treatment with solution B × proportion of B in M

[0056] Toxicity ratio of mixture M = (theoretical mortality rate / actual mortality rate) × 100

[0057] Judgment criteria: toxicity ratio ≥ 1.2 is defined as synergistic effect, 0.8 ≤ toxicity ratio ≤ 1.0 is defined as additive effect, and toxicity ratio < 0.8 is defined as antagonistic effect. The results are shown in Table 3-5 and Figure 1-6 shown.

[0058] Table 3 Contact toxicity test results of cinnamon essential oil solution / (1-naphthyl)-N-methylcarbamate solution ratio on Dermanyssus gallinae

[0059]

[0060] From the data in Table 3, it can be seen intuitively that the theoretical lethality of the combination of cinnamon essential oil solution and (1-naphthyl)-N-methylcarbamate solution ranges from 48.3% to 51.5%. Figure 1 The trend shown is that, starting from Example 2, as the proportion of cinnamon essential oil in the combination gradually increases, the overall acaricidal activity shows a trend of gradually decreasing. In particular, when the volume proportion of cinnamon essential oil reaches 50% (Example 5), the overall acaricidal activity drops to the lowest point, which is only 31.5%. On the contrary, as the content of (1-naphthyl)-N-methylcarbamate solution increases, the acaricidal activity of the combination is enhanced. Figure 3In the combination of cinnamon essential oil solution and (1-naphthyl)-N-methylcarbamate solution, Example 2 has the highest mortality rate, reaching 60%, followed by Example 1, with a mortality rate of 48.3%. The measured mortality rate of Example 5 is the lowest, and its mortality rate is almost half that of Example 2, which has the best combined toxicity performance. In order to quantify the synergistic effect between the test groups, we introduced the indicator of toxicity-effect ratio. The analysis results show that Example 2 has the largest toxicity-effect ratio, as high as 1.22, indicating that this combination exhibits the strongest synergistic effect in the acaricidal effect. The toxicity-effect ratios of Example 1, Example 3 and Example 9 are between 0.8 and 1.2, and these combinations belong to the category of additive effect. As for other combinations, their combined effect is not good, and the toxicity-effect ratio is lower than 0.8, showing antagonism, and their acaricidal activity is significantly lower than that of Example 2, which performs best.

[0061] Table 4 Test results of contact toxicity of oregano essential oil solution / (1-naphthyl)-N-methylcarbamate solution to Dermanyssus gallinae

[0062]

[0063]

[0064] Note: The measured mortality rate, theoretical mortality rate and toxicity-effect ratio data are the average values ​​obtained from four repeated experiments.

[0065] From the data analysis in Table 4 and Figure 2 In the intuitive display, we can clearly see that the theoretical mortality rate of the combination of oregano essential oil solution and (1-naphthyl)-N-methylcarbamate solution is stable between 51.8% and 53.3%. However, in terms of the measured mortality rate, it presents a unique "W"-shaped trend as the content of the oregano essential oil solution changes. Specifically, when the oregano essential oil solution accounts for 60% of the total volume (Example 15), the measured mortality rate peaks at 66.7%; followed by Example 16, which has a mortality rate of 56.7%; and Example 12 performs the worst, with a measured mortality rate of only 33.5%. A toxicity-effect ratio of more than 1.2 indicates the presence of significant synergistic effects. According to Figure 4 According to the data, the toxicity-effect ratio of Example 15 group is as high as 1.27, which is significantly higher than that of other test groups, and this difference is statistically significant (P<0.05), so we can judge that Example 15 group shows the strongest synergistic effect in acaricide toxicity and is regarded as the best test group. In addition, except for Example 12 and Example 13, whose toxicity-effect ratio is lower than 0.8, showing antagonism, the other groups all show the characteristics of additive effect, that is, their toxicity-effect ratio is between 0.8 and 1.2.

[0066] Table 5 Contact toxicity test results of cinnamon essential oil solution / oregano essential oil solution ratio on Dermatosus gallinae

[0067]

[0068] Note: The measured mortality rate, theoretical mortality rate and toxicity-effect ratio data are the average values ​​obtained from four repeated experiments.

[0069] from Figure 5 We can clearly observe that after the cinnamon essential oil solution is mixed with another component, its acaricidal activity shows a unique "S" curve trend as the content of the cinnamon essential oil solution increases, that is, it first decreases, then increases, and finally decreases again. It is worth noting that those combinations that show additive effects are mainly concentrated in the test groups with higher content of cinnamon essential oil solution (Examples 23 to 27). In particular, Example 24, Example 25 and Example 26 in the compound essential oil preparations are particularly outstanding in the acaricidal effect, and their actual measured mortality rates all exceed the theoretical mortality rate, with the highest measured mortality rate reaching 65.3%. In contrast, the measured mortality rates of other test groups are generally lower than the theoretical mortality rate, and the lowest Example 21 even reached only 30%. Although most combinations show certain synergistic or additive effects, there are still some combinations that show antagonistic effects. For example Figure 6 As shown, Example 20 (toxicity ratio 0.77), Example 21 (toxicity ratio 0.65) and Example 22 (toxicity ratio 0.78) are such examples, and their acaricidal activities are significantly lower than that of the best performing combination group of Example 25 (P<0.05).

[0070] 1.3 Fumigation toxicity of examples and comparative examples

[0071] The fumigation toxicity of the binary combination to chicken mites was determined in a closed system. 100 μL of the mixed agent prepared by volume ratio was evenly spread on a filter paper, dried, curled and placed in a 15 mL centrifuge tube. 15 adult female mites were picked and placed in a plastic tube with openings at both ends and the openings were plugged with fluffy absorbent cotton so that the drug vapor could diffuse into the tube and the mites could not escape. The plastic tube with mites was placed in a 15 mL centrifuge tube containing a pretreated filter paper, the lid was closed, and the mortality rate of mites in each time period was observed and recorded. The internal plastic tube was gently shaken for observation, and it was found that the insect body was considered dead if it shook with the tube, the limbs were curled up, and it was unable to crawl. For each compound combination, 9 test groups (Examples 1-27) and three single-dose controls (Comparative Examples 1-3) were set up by volume ratio. At the same time, a solvent was selected as a control. The solvent of the essential oil compound group was ethanol, and the solvent of the essential oil acaricide compound group was acetone. The test results are shown in Tables 6-8 and Figure 7-9 shown.

[0072] Table 6 Mite mortality data of fumigation test with cinnamon essential oil solution / (1-naphthyl)-N-methylcarbamate solution ratio

[0073] sample 6h 12h 24h 36h 48h Comparative Example 3 0％ 0％ 4％ 9％ 20％ Example 1 2％ 7％ 9％ 18％ 22％ Example 2 4％ 0％ 4％ 11％ 18％ Example 3 0％ 0％ 4％ 13％ 22％ Example 4 0％ 0％ 9％ 16％ 33％ Example 5 2％ 4％ 9％ 29％ 38％ Example 6 0％ 7％ 15％ 29％ 51％ Example 7 4％ 7％ 27％ 40％ 64％ Example 8 7％ 22％ 51％ 67％ 82％ Example 9 0％ 33％ 80％ 91％ 98％ Comparative Example 1 0％ 44％ 87％ 100％ 100％

[0074] Note: The displayed data are the average of four test results.

[0075] from Figure 7 As can be seen from Table 6, in the fumigation test conducted in a closed system, as time goes by, the content of essential oil vapor in the centrifuge tube increases significantly, which leads to a steady increase in acaricidal activity. This trend is particularly significant in the combined test of cinnamon essential oil and (1-naphthyl)-N-methylcarbamate. Specifically, when the test is carried out to 48h, the performance of each test group is significantly differentiated. Among them, Example 9 and the pure cinnamon essential oil group (Comparative Example 1) show a significant acaricidal effect, with a caricidal rate of up to 98%, almost killing all mites. In contrast, the single-dose control (1-naphthyl)-N-methylcarbamate group (Comparative Example 3) performed mediocrely, with a mortality rate of only 20%, and the acaricidal rate of Example 2 was also low, only 18%. At 36h of the fumigation test, the pure cinnamon essential oil group had achieved a 100% mite lethality, which proves the potential of cinnamon essential oil to efficiently kill mites in a closed environment. In addition, Example 2, which had the best acaricide effect in the contact test, showed relatively low toxicity in the fumigation test, while the compound groups with high fumigation toxicity, such as Example 8 (82% lethality) and Example 9 (98% lethality), performed poorly in the contact test.

[0076] Table 7 Mite mortality data of oregano essential oil solution / (1-naphthyl)-N-methylcarbamate solution ratio fumigation test

[0077] sample 6h 12h 24h 36h 48h Comparative Example 3 0％ 0％ 0％ 2％ 16％ Example 10 2％ 2％ 2％ 7％ 16％ Embodiment 11 4％ 7％ 18％ 24％ 36％ Example 12 0％ 0％ 22％ 27％ 38％ Embodiment 13 0％ 13％ 33％ 38％ 53％ Embodiment 14 2％ 7％ 29％ 49％ 62％ Embodiment 15 11％ 47％ 64％ 76％ 96％ Example 16 29％ 49％ 67％ 89％ 100％ Embodiment 17 20％ 58％ 87％ 96％ 100％ Embodiment 18 36％ 78％ 100％ 100％ 100％ Comparative Example 2 40％ 87％ 100％ 100％ 100％

[0078] Note: The displayed data are the average of four test results.

[0079] As shown in Table 7, in the compounding test of oregano essential oil solution and (1-naphthyl)-N-methylcarbamate solution, each embodiment showed a significant mite killing effect. After 48 hours of treatment, the mite mortality rate treated by Example 15 reached 96%, while Examples 16-18 achieved a 100% mite lethality. Further observations showed that when the treatment time was shortened to 24 hours, the lethality of the compounding group of Example 15 was as high as 64%, while that of the compounding group of Example 16 was 67%. It is worth noting that despite the increase of 10% in the dose of essential oil solution, the lethality of Example 16 only increased by 2% compared with Example 15, which indicates that within this concentration range, the mite killing effect is not proportional to the increase in dose, but there is a certain saturation effect. At the same time, the pure oregano essential oil solution group had reached a lethality of 100% at 24 hours, highlighting the powerful mite killing potential of oregano essential oil in a short period of time. Figure 8It is clearly shown that the four compound ratios of Example 15, Example 16, Example 17 and Example 18 all exhibited excellent fumigation mite killing activity, and were the most outstanding combinations in this test.

[0080] From the above two groups of fumigation results, it can be seen that based on their respective LC 50 The fumigation toxicity of oregano essential oil solution to Dermanyssus gallinae is higher than that of cinnamon essential oil, and a 60% dosage of oregano essential oil mixed with (1-naphthyl)-N-methylcarbamate solution can achieve a 96% mite killing rate.

[0081] Table 8 Cinnamon essential oil solution / oregano essential oil solution ratio fumigation test mite mortality data

[0082] sample 6h 12h 18h 24h 30h Comparative Example 2 4％ 100％ 98％ 100％ 100％ Embodiment 19 7％ 87％ 98％ 100％ 100％ Embodiment 20 16％ 88％ 97％ 100％ 100％ Embodiment 21 0％ 57％ 85％ 88％ 100％ Embodiment 22 0％ 40％ 95％ 96％ 96％ Embodiment 23 2％ 21％ 67％ 80％ 95％ Embodiment 24 0％ 8％ 78％ 83％ 87％ Embodiment 25 2％ 24％ 63％ 72％ 85％ Embodiment 26 0％ 15％ 53％ 79％ 91％ Embodiment 27 0％ 17％ 58％ 73％ 89％ Comparative Example 1 0％ 18％ 70％ 88％ 98％

[0083] Note: The displayed data are the average of four test results.

[0084] From Table 8 and Fig. 9 It can be seen that each compound group of the essential oil combination reached a high mortality rate after 30 hours of the test. Among them, the mortality rate of the three compound groups of Example 19-Example 21 reached 100%, the mortality rates of the compound groups of Example 22, Example 23 and Example 26 were 96%, 95% and 91% respectively, and the mortality rates of the three compound groups of Example 24, Example 25 and Example 27 were 87%, 85% and 89%. When the test was carried out for 12 hours, the mortality rate of mites treated by the compound groups of Example 19 and Example 20 reached 87%, while that of Example 22 was only 40%; it is worth noting that when the test was carried out for 18 hours, the three compound combinations all reached a mortality rate of 95%. Although the mortality rate of mites in the compound group of Example 25 was not good in the first 12 hours of the test, it reached a mortality rate of 85% at 30 hours of the test, indicating that it can effectively kill mites. All compounding ratios of the entire essential oil solution combination have good fumigation toxicity to gallinae.

[0085] 1.4 Examples and Comparative Examples Ovicidal Activity

[0086] The composite agents obtained in Example 2, Example 15, Example 25 and Comparative Examples 1, 2, and 3 were subjected to an egg hatching experiment. Take 100 μL of the drug solution and evenly apply it on a filter paper with a diameter of 35 mm. After drying, put it in a culture dish and pick up 15 eggs. Place the culture dish in an incubator at a temperature of 30°C and a relative humidity of 70%. Observe the hatching of the eggs on the third day and record the number of unhatched eggs. The criteria for judging the death of eggs are as follows: the surface of the eggs is obviously wrinkled, the color is brownish yellow or there is no obvious change in the appearance of the eggs. At the same time, the eggs that have not hatched larvae after 72 hours of the test are considered dead. Set up two solvent control groups of acetone and ethanol and a distilled water blank control group.

[0087] Table 9 Egg hatching test results

[0088]

[0089] Note: The adjusted mortality rate is the average of three replicates.

[0090] The results of the egg hatching test of Dermatosus gallinae treated with the optimal ratio of each compound combination are shown in Table 9 and Figure 10-11 As shown in Table 9, the mortality rate of the eggs after treatment with the composite agent obtained in Example 2 reached 98%, the mortality rate of the eggs after treatment with the composite agent obtained in Example 15 reached 94%, and the mortality rate of the eggs after treatment with the composite agent obtained in Example 25 reached 96%. The mortality rates of the eggs after treatment with the three single agents in Comparative Examples 1-3 were 69%, 67% and 80%, respectively. The hatching rate of the eggs after treatment with the three best compound combinations was significantly reduced compared with that of the comparative example and the control group. The results are shown in Fig.10 As shown, the composite agents provided in Example 2, Example 15, and Example 25 can effectively inhibit the hatching of eggs. Among them, the hatching rate of eggs after treatment with the composite agent obtained in Example 2 is the lowest, which is only 2%, and the difference is statistically significant (P<0.05). Fig.11 The hatching test was carried out for 48 hours, and the AC graphs were respectively for Example 2, Example 15, and Example 25; the DF graphs were respectively for Comparative Example 1, Comparative Example 2, and Comparative Example 3; and the GI graphs were respectively for the acetone control group, the ethanol control group, and the distilled water blank control group. Observing the egg status of each group and combining the hatching results in Table 9, it can be seen that after being treated with the binary mixture group (A, B, C) and the single agent group (D, E, F), the surface of the mite eggs was obviously wrinkled, with obvious depressions and protrusions, and the color deepened to brownish yellow, and the internal structure could not be distinguished. The egg structure was destroyed, so the hatching rate was low; correspondingly, after being treated with the control group (G, H, I), the surface of the eggs was smooth and transparent, the internal tissue was clearly visible and the development state was good, and the egg structure was complete, so the hatching rate was significantly higher than that of the other two groups.

[0091] According to the above test results, we conducted a contact test based on the LC of each solution. 50 The optimal ratios of three binary compound combinations of cinnamon essential oil solution and (1-naphthyl)-N-methylcarbamate solution, oregano essential oil solution and (1-naphthyl)-N-methylcarbamate solution, and cinnamon essential oil solution and oregano essential oil solution were screened out by volume ratio. Further evaluation and verification through fumigation test and insect egg hatching test, we came to the following conclusions: The optimal compound ratio of cinnamon essential oil and (1-naphthyl)-N-methylcarbamate is 20% by volume of cinnamon essential oil LC 50 Solution and 80% volume (1-naphthyl)-N-methylcarbamate LC 50Solution; the optimal compounding ratio of oregano essential oil and (1-naphthyl)-N-methylcarbamate is 60% by volume oregano essential oil LC 50 Solution and 40% volume (1-naphthyl)-N-methylcarbamate LC 50 solution; while the binary mixture of cinnamon essential oil and oregano essential oil was 70% by volume cinnamon essential oil LC 50 Solution with 30% volume oregano essential oil LC 50 The solution is the best compound ratio. These combinations have shown significant effects in killing or inhibiting the activity of mites in terms of contact toxicity, fumigation toxicity and egg hatching inhibition.

[0092] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modification, equivalent substitution or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A pharmaceutical composition for killing Dermanyssus gallinae, characterized in that: The pharmaceutical composition comprises at least two of cinnamon essential oil, oregano essential oil or carbamate compounds.

2. The pharmaceutical composition for killing Dermanyssus gallinae according to claim 1, characterized in that: The pharmaceutical composition consists of cinnamon essential oil and oregano essential oil; or the pharmaceutical composition consists of cinnamon essential oil and carbamate compounds; or the pharmaceutical composition consists of oregano essential oil and carbamate compounds.

3. The pharmaceutical composition for killing Dermanyssus gallinae according to claim 1 or 2, characterized in that: The carbamate compound is (1-naphthyl)-N-methylcarbamate.

4. A pharmaceutical preparation for killing Dermanyssus gallinae, characterized in that: The pharmaceutical preparation comprises the pharmaceutical composition for killing Dermanyssus gallinae according to any one of claims 1 to 3.

5. The pharmaceutical preparation for killing Dermanyssus gallinae according to claim 4, characterized in that: The pharmaceutical preparation also includes a diluent.

6. The pharmaceutical preparation for killing Dermanyssus gallinae according to claim 4, characterized in that: When the pharmaceutical composition consists of cinnamon essential oil and oregano essential oil, the final concentration of cinnamon essential oil in the pharmaceutical preparation is 14 mg / mL-22 mg / mL, and the final concentration of oregano essential oil is 2 mg / mL-12 mg / mL.

7. The pharmaceutical preparation for killing Dermanyssus gallinae according to claim 4, characterized in that: When the pharmaceutical composition consists of cinnamon essential oil and carbamate compounds, the final concentration of cinnamon essential oil in the pharmaceutical preparation is 2 mg / mL-8 mg / mL, and the final concentration of carbamate compounds is 7 μg / mL-9 μg / mL.

8. The pharmaceutical preparation for killing Dermanyssus gallinae according to claim 4, characterized in that: When the pharmaceutical composition is composed of oregano essential oil and carbamate compounds, the final concentration of oregano essential oil in the pharmaceutical preparation is 14 mg / mL-26 mg / mL, and the final concentration of carbamate compounds is 1 μg / mL-5 μg / mL.

9. The pharmaceutical preparation for killing Dermanyssus gallinae according to claim 4, characterized in that: The diluent is acetone.

10. The method for preparing the pharmaceutical preparation for killing Dermanyssus gallinae according to any one of claims 4 to 9, characterized in that: The steps include: S1, dissolving cinnamon essential oil, oregano essential oil and carbamate compound in diluent respectively to obtain cinnamon essential oil solution, oregano essential oil solution and carbamate compound solution; S2. Mix at least two of the cinnamon essential oil solution, the oregano essential oil solution or the carbamate compound solution to obtain a pharmaceutical preparation for killing Dermanyssus gallinae.

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