MITE INFESTATION TREATMENT

Abstract

The invention relates to a compound of formula (I), a salt thereof or a composition containing same as an acaricide, a method for reducing or preventing an infestation of an animal by a mite, comprising exposing the mite to a compound of formula (I), to a composition comprising a compound of formula (I), to one or more attractants for bees and a polymer, to a strap adapted for use in apiculture comprising a compound of formula (I) and to a hive comprising a compound of formula (I).

Claims

1-2. (canceled)

3. A method for reducing or preventing mite infestation of an animal, comprising exposing the mite to a compound of formula (I), a salt thereof or a composition containing same, said formula (I) is: ##STR00033## wherein: A is selected from the group (II) or (III): ##STR00034## X is NH, CH.sub.2 or CH—CH.sub.3; Y is a 5-bonded heterocycle comprising at least one nitrogen atom; Z.sub.1 is a halogen, H or CH.sub.3; Z.sub.2 is a halogen, H or CH.sub.3; Z.sub.3 is a halogen, H or CH.sub.3; Z.sub.4 is a halogen, H or CH.sub.3.

4. The method according to claim 3, wherein the composition of the compound of formula (I) or the composition of the salt of the compound of formula (I) contains from 0.001 to 200 μg/μL of compound of formula (I) or salt thereof, preferably from 0.01 to 100 μg/μL.

5. The method according to claim 3, wherein the mite infestation of an animal is a mite infestation of a beehive, preferably a beehive of the Apis mellifera, Apis cerana, Apis dorsata or Apis florea genus.

6. The method according to claim 3, wherein the reduction in mite infestation is a reduction of at least 25%, preferably at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or even 100% of the number of live mites attached to the animal.

7. The method according to claim 3, wherein the compound of formula (I), the salt thereof or the composition containing same is comprised in and/or on a strap.

8. The method according to claim 3, wherein the animal is a bee and the mite is Varroa mite.

9. A composition adapted for use as a Varroacid in bees, said composition comprising a compound of formula (I) or a salt thereof, one or more attractant(s) for bees, and one or more polymer(s) selected from a plastic material, a rubber, an adhesive, a resin and polyholoside fibers, said formula (I) is: ##STR00035## wherein: A is selected from the group (II) or (III): ##STR00036## X is NH, CH.sub.2 or CH—CH.sub.3; Y is a 5-bonded heterocycle comprising at least one nitrogen atom; Z.sub.1 is a halogen, H or CH.sub.3; Z.sub.2 is a halogen, H or CH.sub.3; Z.sub.3 is a halogen, H or CH.sub.3; Z.sub.4 is a halogen, H or CH.sub.3.

10. A strap adapted for use in apiculture, comprising (i) a compound of formula (I), a salt thereof, a composition containing same or (ii) a composition according to claim 9, said formula (I) is: ##STR00037## wherein: A is selected from the group (II) or (III): ##STR00038## X is NH, CH.sub.2 or CH—CH.sub.3; Y is a 5-bonded heterocycle comprising at least one nitrogen atom; Z.sub.1 is a halogen, H or CH.sub.3; Z.sub.2 is a halogen, H or CH.sub.3; Z.sub.3 is a halogen, H or CH.sub.3; Z.sub.4 is a halogen, H or CH.sub.3.

11. A hive comprising (i) a compound of formula (I), a salt thereof or a composition containing same, (ii) a composition according to claim 9, or (iii) a strap according to claim 10 adapted for use in apiculture, comprising the compound of (i) or the composition of (ii), said formula (I) is: ##STR00039## wherein: A is selected from the group (II) or (III): ##STR00040## X is NH, CH.sub.2 or CH—CH.sub.3; Y is a 5-bonded heterocycle comprising at least one nitrogen atom; Z.sub.1 is a halogen, H or CH.sub.3; Z.sub.2 is a halogen, H or CH.sub.3; Z.sub.3 is a halogen, H or CH.sub.3; Z.sub.4 is a halogen, H or CH.sub.3.

12. The hive according to claim 11, wherein Y is the group (IV) or (V): ##STR00041##

13. The hive according to claim 11, wherein: A is the group (II), X is NH, Y is the group (IV), Z.sub.1 is a halogen, Z.sub.2 is H and Z.sub.3 is a halogen, A is the group (II), X is CH.sub.2 or CH—CH.sub.3, Y is the group (V), Z.sub.1 is CH.sub.3, Z.sub.2 is CH.sub.3 and Z.sub.3 is H, or A is the group (III), X is NH, Y is the group (IV) and Z.sub.4 is a halogen.

14. The hive according to claim 11, wherein the allogen is selected from F, Cl or Br.

15. The hive according to claim 11, wherein the compound of formula (I) is selected from detomidine, dexmedetomidine, medetomidine, romifidine, clonidine, tizanidine or a salt of one of these compounds.

16. The method according to claim 3, wherein Y is the group (IV) or (V): ##STR00042##

17. The method according to claim 3, wherein: A is the group (II), X is NH, Y is the group (IV), Z.sub.1 is a halogen, Z.sub.2 is H and Z.sub.3 is a halogen, A is the group (II), X is CH.sub.2 or CH—CH.sub.3, Y is the group (V), Z.sub.1 is CH.sub.3, Z.sub.2 is CH.sub.3 and Z.sub.3 is H, or A is the group (III), X is NH, Y is the group (IV) and Z.sub.4 is a halogen.

18. The method according to claim 3, wherein the allogen is selected from F, Cl or Br.

19. The method according to claim 3, wherein the compound of formula (I) is selected from detomidine, dexmedetomidine, medetomidine, romifidine, clonidine, tizanidine or a salt of one of these compounds.

20. The composition according to claim 9, wherein Y is the group (IV) or (V): ##STR00043##

21. The composition according to claim 9, wherein: A is the group (II), X is NH, Y is the group (IV), Z.sub.1 is a halogen, Z.sub.2 is H and Z.sub.3 is a halogen, A is the group (II), X is CH.sub.2 or CH—CH.sub.3, Y is the group (V), Z.sub.1 is CH.sub.3, Z.sub.2 is CH.sub.3 and Z.sub.3 is H, or A is the group (III), X is NH, Y is the group (IV) and Z.sub.4 is a halogen.

22. The composition according to claim 9, wherein the allogen is selected from F, Cl or Br.

23. The composition according to claim 9, wherein the compound of formula (I) is selected from detomidine, dexmedetomidine, medetomidine, romifidine, clonidine, tizanidine or a salt of one of these compounds.

24. The strap according to claim 10, wherein Y is the group (IV) or (V): ##STR00044##

25. The strap according to claim 10, wherein: A is the group (II), X is NH, Y is the group (IV), Z.sub.1 is a halogen, Z.sub.2 is H and Z.sub.3 is a halogen, A is the group (II), X is CH.sub.2 or CH—CH.sub.3, Y is the group (V), Z.sub.1 is CH.sub.3, Z.sub.2 is CH.sub.3 and Z.sub.3 is H, or A is the group (III), X is NH, Y is the group (IV) and Z.sub.4 is a halogen.

26. The strap according to claim 10, wherein the allogen is selected from F, Cl or Br.

27. The strap according to claim 10, wherein the compound of formula (I) is selected from detomidine, dexmedetomidine, medetomidine, romifidine, clonidine, tizanidine or a salt of one of these compounds.

28. An acaricide composition containing a compound of formula (I), or a salt thereof, said formula (I) is: ##STR00045## wherein: A is selected from the group (II) or (III): ##STR00046## X is NH, CH.sub.2 or CH—CH.sub.3; Y is a 5-bonded heterocycle comprising at least one nitrogen atom; Z.sub.1 is a halogen, H or CH.sub.3; Z.sub.2 is a halogen, H or CH.sub.3; Z.sub.3 is a halogen, H or CH.sub.3; Z.sub.4 is a halogen, H or CH.sub.3.

29. The acaricide composition according to claim 28, wherein said composition reduces or prevents an infestation of an animal by a mite.

30. The acaricide composition according to claim 28, wherein Y is the group (IV) or (V): ##STR00047##

31. The acaricide composition according to claim 28, wherein: A is the group (II), X is NH, Y is the group (IV), Z.sub.1 is a halogen, Z.sub.2 is H and Z.sub.3 is a halogen, A is the group (II), X is CH.sub.2 or CH—CH.sub.3, Y is the group (V), Z.sub.1 is CH.sub.3, Z.sub.2 is CH.sub.3 and Z.sub.3 is H, or A is the group (III), X is NH, Y is the group (IV) and Z.sub.4 is a halogen.

32. The acaricide composition according to claim 28, wherein the allogen is selected from F, Cl or Br.

33. The acaricide composition according to claim 28, wherein the compound of formula (I) is selected from detomidine, dexmedetomidine, medetomidine, romifidine, clonidine, tizanidine or a salt of one of these compounds.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0102] FIG. 1A-1B is a diagram which represents the toxicity of certain solvents towards Varroa mites and bees. The negative control corresponds to untreated bees, that is to say having received no thoracic deposition.

[0103] FIG. 2A-2B is a diagram that measures the Varroacid effect of amitraz (FIG. 2A) and its effect on bees (FIG. 2B). The negative control corresponds to untreated bees, that is to say having received no thoracic deposition. The solvent allowed to solubilize the molecule and/or to carry out dilutions or concentrations.

[0104] FIG. 3A-3B is a diagram that measures the Varroacid effect of clonidine hydrochloride (FIG. 3A) and its effect on bees (FIG. 3B). The negative control corresponds to untreated bees, that is to say having received no thoracic deposition. The solvent allowed to solubilize the molecule and/or to carry out dilutions or concentrations.

[0105] FIG. 4A-4B is a diagram that measures the Varroacid effect of Detomidine hydrochloride monohydrate (FIG. 4A) and its effect on bees (FIG. 4B). The negative control corresponds to untreated bees, that is to say having received no thoracic deposition. The solvent allowed to solubilize the molecule and/or to carry out dilutions or concentrations.

[0106] FIG. 5A-5B is a diagram that measures the Varroacid effect of dexmedetomidine hydrochloride (FIG. 5A) and its effect on bees (FIG. 5B). The negative control corresponds to untreated bees, that is to say having received no thoracic deposition. The solvent allowed to solubilize the molecule and/or to carry out dilutions or concentrations.

[0107] FIG. 6A-6B is a diagram that measures the Varroacid effect of romifidine hydrochloride (FIG. 6A) and its effect on bees (FIG. 6B). The negative control corresponds to untreated bees, that is to say having received no thoracic deposition. The solvent allowed to solubilize the molecule and/or to carry out dilutions or concentrations.

[0108] FIG. 7A-7B is a diagram that measures the Varroacid effect of tizanidine hydrochloride (FIG. 7A) and its effect on bees (FIG. 7B). The negative control corresponds to untreated bees, that is to say having received no thoracic deposition. The solvent allowed to solubilize the molecule and/or to carry out dilutions or concentrations.

[0109] FIG. 8A-8B is a diagram that measures the Varroacid effect of medetomidine hydrochloride (FIG. 8A) and its effect on bees (FIG. 8B). The negative control corresponds to untreated bees, that is to say having received no thoracic deposition. The solvent allowed to solubilize the molecule and/or to carry out dilutions or concentrations.

[0110] FIG. 9 represents the Varroacid effect of dexmedetomidine hydrochloride by bee feeding. The negative control corresponds to bees receiving only pure syrup.

[0111] FIG. 10A-10B is a diagram that measures the Varroacid effect of levmedetomidine hydrochloride (FIG. 10A) and its effect on bees (FIG. 10B). The negative control corresponds to untreated bees, that is to say having received no thoracic deposition. The solvent allowed to solubilize the molecule and/or to carry out dilutions or concentrations.

[0112] FIG. 11A-11B is a diagram that measures the Varroacid effect of Octopamine (FIG. 11A) and its effect on bees (FIG. 11B). The negative control corresponds to untreated bees, that is to say having received no thoracic deposition. The solvent allowed to solubilize the molecule.

[0113] FIG. 12 is a photograph showing feeding units for rearing ticks.

[0114] FIG. 13 is a diagram representing the percentage of dead ticks according to the different treatments over a period of 7 days.

[0115] FIG. 14 is a diagram that measures the cumulative efficiency of the tested compounds as a function of treatment time against Varroa mites in beehives.

[0116] FIG. 15 is a diagram that measures the mortality of adult bees following the application of the compounds tested in beehives.

EXAMPLES

Example 1: Demonstration of the Acaricidal Effect of the Compound of Formula (I) Against Varroa destructor

[0117] Materials and Methods

[0118] Preparation of Varroa Mites

[0119] About 300 bees (Apis mellifera) infected with Varroa destructor were taken from a hive superinfected with Varroa destructor. The bees were then placed in a device allowing to detach Varroa mites attached to the bees (device described in Community model n° 003419415-0001) filled with icing sugar. The device was stirred for 2 minutes to sample the Varroa mites attached to the bees. The icing sugar allows to interfere between the bee and the Varroa mite that was attached thereto. Varroa mites were harvested with icing sugar because they pass through the holes in the device, unlike bees. The Varroa mites were then separated from the icing sugar.

[0120] The Varroa mites thus recovered were dispatched in boxes (10 Varroa mites per box). Two boxes were used to test each compound at the same dose, in order to have data for 20 Varroa mites.

[0121] Preparation of the Bees

[0122] The bees were harvested from hives which had not undergone any treatment during the last 6 months in order to be sure that the effect of the experiments did not come from a previous treatment.

[0123] The bees were then dispatched in the boxes (10 per box). Two boxes were used to test the same compound at the same dose, in order to have data for 20 bees.

[0124] The bees were then put to sleep for a few minutes using CO.sub.2 (flow of 3 liters for 20 seconds) in order to deposit 1 μL of compound to be tested on their thorax using a pipette. It should be noted that the bees which received romifidine hydrochloride at a concentration of 10 μg/μL received 2 μL of product. The bees were unconscious for the time the compound to be tested was deposited, that is to say around three minutes, then they woke up.

[0125] After thoracic deposition of the compound to be tested, the 10 bees from the same box were transferred to a box containing the 10 Varroa mites (cf. “Preparation of Varroa mites” above). Liquid sugar was left in each of the boxes to feed the bees during the observations. The series of boxes was then placed in an oven at 25° C. with a humidity of 50% in order to approximate hive conditions as closely as possible. It is made sure that after 5 minutes all the Varroa mites were attached to the bees.

[0126] Effect of the Compound on Varroa Mites and the Bees

[0127] The mortality of Varroa mites and bees was then monitored 5 minutes, 2 hours, 4 hours and 24 hours after application of the compound to be tested to the bees. Dead Varroa mites correspond to Varroa mites detached from the bees. Dead bees correspond to bees which no longer move and which are often placed on their thorax.

[0128] Test of the Solvents Used

[0129] The solvents were tested in order to be sure that they had no (positive or negative) impact on the results. Four solvents were tested: ethanol, methanol, acetone and acetonitrile.

[0130] Results

[0131] The results are shown in FIG. 1. The solvents tested did not prove to be significantly toxic either for Varroa mites or for bees.

[0132] Conclusion

[0133] The solvents tested had no impact on the viability of bees and Varroa mites. They can therefore be used to dilute the tested compounds.

[0134] Test of the Compounds

[0135] Tested Compounds

[0136] The following compositions were applied to the thorax of the bees: [0137] Solvent, [0138] Amitraz at 1 μg/μL in acetone, [0139] Clonidine hydrochloride at 100 μg/μL, 10 μg/μL, 1 μg/μL and 0.1 μg/μL in methanol, [0140] Detomidine hydrochloride monohydrate at 1 μg/μL in methanol, [0141] Dexmedetomidine hydrochloride at 100 μg/μL, 10 μg/μL, 1 μg/μL and 0.1 μg/μL in methanol, [0142] Romifidine hydrochloride at 10 μg/μL and at 5 μg/μL in methanol, [0143] Tizanidine hydrochloride at 1 μg/μL in methanol, [0144] Medetomidine hydrochloride at 0.1 μg/μL in ethanol, [0145] Levomedetomidine hydrochloride at 100 μg/μL, 10 μg/μL, 1 μg/μL and 0.1 μg/μL in methanol, [0146] Octopamine at 1 μg/μL in ethanol.

[0147] Results

[0148] Amitraz: 100% of the Varroa mites were killed and 0% of the bees were killed (FIG. 2). As expected, amitraz is very effective in the fight against Varroa mites. Amitraz is a positive control.

[0149] Clonidine hydrochloride: the results are presented in FIG. 3. At concentrations of 100 μg/μL and 10 μg/μL, the mortality of Varroa mites reached 85% for only 10% of the bees killed. Surprisingly, the efficiency of the compound against Varroa mites was particularly marked at the concentration of 1 μg/μL with zero mortality for bees and 95% mortality for Varroa mites. The kinetics of this molecule appeared rather slow compared to the positive control because it showed its full efficiency after 24 hours.

[0150] Detomidine hydrochloride monohydrate: the results are presented in FIG. 4. Detomidine hydrochloride monohydrate proved to be particularly effective since after 2 hours, 40% of the Varroa mites were killed without mortality for the bees. After 24 hours, 100% of Varroa mites were killed without mortality for the bees.

[0151] Dexmedetomidine hydrochloride: the results are presented in FIG. 5. The results revealed that only the concentration of 100 μg/μL is toxic for bees. At other concentrations (10 μg/μl; 1 μg/μl; 0.1 μg/μl) mortality of less than 5% for bees, and systematically 100% mortality for Varroa mites after 24 hours were observed. Varroa was therefore sensitive to this compound.

[0152] Romifidine hydrochloride: the results are presented in FIG. 6. The molecule proved to be very effective. After 2 hours after depositing 2 μl of product at 10 μg/μL, the mortality of Varroa mites was 75% then 95% after 4 hours. For the concentration at 5 μg/μL, it took 24 hours to obtain the same level of mortality for Varroa mites. Bee mortality remained zero at all doses and even after 24 hours (diagram not shown).

[0153] Tizanidine hydrochloride: the results are shown in FIG. 7. The product proved to be very effective at 1 μg/μL after 24 hours (80% Varroa mite mortality) without toxicity for bees.

[0154] Medetomidine hydrochloride: the results are shown in FIG. 8. The product proved to be very effective.

[0155] Levomedetomidine hydrochloride: the results are shown in FIG. 10. The results are similar to those obtained with dexmedetomidine. Varroa mites were therefore sensitive to this compound.

[0156] Octopamine: the results are shown in FIG. 11. No Varroacid effect was observed. Bee mortality also remained zero.

Example 2: Exposure to the Compound of Formula (I) by Feeding

[0157] Bees and Varroa mites were obtained and contacted as detailed in Example 1 (without the treatment step).

[0158] The bees on which the Varroa mites are attached (10 bees placed in contact with 10 Varroa mites) were fed with: [0159] A pure syrup (Negative control), or [0160] A syrup comprising dexmedetomidine hydrochloride at 100 μg/mL syrup.

[0161] The syrup was provided to the bees ad libitum in 1.5 mL eppendorf tubes.

[0162] Results

[0163] The results are presented in FIG. 9 for the Varroacid effect and in Table 1 for the toxicity on bees.

TABLE-US-00001 TABLE 1 % bee mortality 5 min 1 h 2 h 3 h 30 3 h 45 24 h Negative control 0 0 0   0   0   0   Dexmedetomidine 0 0 5% 5% 5% 5% 100 μg/mL syrup

Example 3: Demonstration of the Acaricidal Effect of Compounds According to the Invention Against Ixodes ricinus

[0164] Compounds Tested According to the Invention

[0165] SN0305: Dexmedetomidine hydrochloride

[0166] SN0306: Medetomidine hydrochloride

[0167] SN0307: Detomidine hydrochloride

[0168] Summary

[0169] A laboratory study was conducted to determine the oral acaricidal properties of three compounds according to the invention (SN0305, SN0306 and SN0307) against Ixodesricinus.

[0170] Each compound was tested at 1, 10 and 100 ppm, and compared to a negative control corresponding to solvent alone (0.1% DMSO). Amitraz at 100 ppm was used as a positive control.

[0171] The Ixodes ricinus ticks (nymphs and adults) were placed for 24 hours on a membrane in feeding wells containing blood mixed with the tested compounds. Tick mortality (number of dead, moribund, or unaffected ticks in each well), attachment to the membrane (number of ticks attached/detached in each well), and the behavior (presence or absence of twitches) were examined 1, 3, 6, 12, 24, 48, 72, and 96 hours after the onset of tick exposure to the compounds.

[0172] Methodology

[0173] Test System—Arthropods

[0174] Ticks (Ixodes ricinus) were collected. Collected adult females were kept separate from the nymphs and both were placed in sterile tubes in a humidity chamber to provide ˜80% humidity and 17° C. Only healthy ticks (females/nymphs) were chosen for the experiment.

[0175] Batches of 12 adults or nymphs were inserted into the feeding wells. After about 24 hours, their attachment was examined with a brush. Dead or moribund ticks and ticks that did not attach were removed.

[0176] Test Article—Compounds

[0177] The efficiency of three compounds (SN0305, SN0306 and SN0307), each tested at three concentrations (1, 10, 100 ppm), was compared to the solvent control (0.1% DMSO, Sigma-Aldrich D-5879; Lot 101K0028) and to the positive control (Amitraze at 100 ppm).

[0178] Membranes

[0179] Silicone-coated membranes with a thickness comprised between 110 and 140 μm were used for the construction of the feeding units.

[0180] Feeding Units

[0181] Feeding units (sterilized acrylic glass tubes) were placed in feeding wells made of sterilized six-well plates.

[0182] 3.1 ml of sterile 10.sup.−3 M ATP-enriched blood was applied to each feeding well. Feeding wells were then inserted into a heated water bath (FIG. 12) and maintained at 37° C.±2° C.

[0183] Tested Compounds and Application

[0184] Stock solutions of 100,000 ppm were prepared and tenth dilutions were carried out in 0.1% DMSO to reach the desired concentrations. After tick attachment (24 hours after placing the ticks in the feeding wells), 3.1 μl of the solutions were added to each feeding well containing 3.1 ml of blood. Blood was changed after 12 hours with fresh supplementation of the compounds to be tested. The ticks were exposed to the product for a total of 24 hours, after which the blood was again changed to fresh, untreated blood.

[0185] Evaluations

[0186] Tick mortality (number of dead, moribund or unaffected ticks in each well), attachment (number of attached/detached ticks in each well) and behavior (presence, absence and intensity of convulsions) were examined 1, 3, 6, 12, 24, 48, 72, and 96 hours after the start of tick exposure to the compound.

[0187] Results

[0188] Convulsion, Morbidity or Death of Ticks

[0189] The three tested compounds (SN0305, SN0306 and SN0307), regardless of the concentration tested, showed a strong effect on ticks from 1 hour of exposure (convulsion, morbidity or death of ticks). 100% of ticks were affected. The amitraz positive control obtained a comparable result for nymphs, but a much weaker effect was observed on adult ticks (Table 2). No effect was observed for the solvent (negative control) during the 96 h of the experiment.

TABLE-US-00002 TABLE 2 average percentage of affected adult ticks (convulsion, morbidity or death of ticks) after the start of oral exposure to amitraz at 100 ppm. Time (h) 1 3 6 12 24 48 72 96 % affected 66.8 56.2 53.7 56.2 66.5 83.1 100.0 100.0

[0190] Detachment of Ticks

[0191] The three products tested (SN0305, SN0306 and SN0307) showed a very limited effect in terms of tick detachment. Only very isolated cases were observed: 1 detached nymph in a replica of SN0305 at 1 ppm, 1 detached adult tick in a replica of SN0305 at 100 ppm, 2 detached adults in a replica of SN0307 at 1 ppm. 55% of fully fed nymphs released naturally between 72 and 96 h in the solvent control. No adult ticks were detached in the solvent control.

[0192] Amitraz has induced a strong detachment of adult ticks (Table 3).

TABLE-US-00003 TABLE 3 average percentage of attached adult ticks after the start of exposure to amitraz at 100 ppm. Time (h) 1 3 6 12 24 48 72 96 % attached 66.8 56.2 53.7 53.7 53.7 53.7 53.7 53.7

[0193] Tick Mortality

[0194] The three products tested (SN0305, SN0306 and SN0307) caused 100% mortality (moribund or dead ticks) in nymphs and adult ticks after 96 h. Amitraz caused 100% mortality in nymphs and 97.5% in adults. No mortality was observed with the solvent (Tables 4 and 5).

TABLE-US-00004 TABLE 4 average percentage of mortality of tick nymphs (moribund or dead nymphs) exposed to SN0305, SN0306 and SN0307 at 1, 10 and 100 ppm, solvent control (0.1% DMSO) or Amitraz at 100 ppm. Conc. Product (ppm) 1 h 3 h 6 h 12 h 24 h 48 h 72 h 96 h 305 1 0.0 0.0 13.8 31.3 58.9 85.7 100.0 100.0 305 10 0.0 9.2 24.9 48.3 83.0 96.9 100.0 100.0 305 100 0.0 13.3 33.0 59.0 92.9 100.0 100.0 100.0 306 1 0.0 0.0 19.5 38.9 61.8 93.9 100.0 100.0 306 10 0.0 10.4 38.4 58.5 78.7 85.0 95.8 100.0 306 100 0.0 9.1 38.5 69.2 94.4 100.0 100.0 100.0 307 1 0.0 0.0 14.1 35.4 56.4 97.2 100.0 100.0 307 10 0.0 12.7 35.5 54.7 88.1 100.0 100.0 100.0 307 100 0.0 9.4 45.3 69.5 95.5 100.0 100.0 100.0 Amitraz 100 0.0 14.0 38.7 74.9 100.0 100.0 100.0 100.0 Solvent 0.1% DMSO 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0

TABLE-US-00005 TABLE 5 average percentage of adult tick mortality (moribund or dead adult ticks) exposed to SN0305, SN0306 and SN0307 at 1, 10 and 100 ppm, solvent (0.1% DMSO) or AMITRAZE at 100 ppm. Survival of adult ticks in amitraz treatment may be caused by early detachment of ticks. Conc, Product (ppm) 1 h 3 h 6 h 12 h 24 h 48 h 72 h 96 h 305 1 0.0 0.0 3.1 3.1 64.2 88.2 100.0 100.0 305 10 0.0 0.0 5.6 11.1 57.3 79.5 96.9 100.0 305 100 0.0 0.0 5.9 11.8 63.9 76.4 91.3 100.0 306 1 0.0 0.0 2.3 7.3 47.6 97.7 100.0 100.0 306 10 0.0 0.0 9.6 9.6 53.9 91.2 100.0 100.0 306 100 0.0 0.0 12.5 18.1 53.1 84.7 93.8 100.0 307 1 0.0 0.0 2.8 2.8 62.9 88.2 100.0 100.0 307 10 0.0 0.0 8.3 8.3 52.1 96.4 100.0 100.0 307 100 0.0 0.0 8.8 15.4 24.0 57.4 100.0 100.0 Amitraz 100 0.0 2.8 10.1 17.4 27.7 58.9 84.6 97.5 solvent 0.1% DMSO 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0

[0195] Conclusion

[0196] All the tested compounds showed an acaricidal effect on Ixodes ricinus.

Example 4: Demonstration of the Acaricidal Effect of Compounds According to the Invention Against Rhipicephalus sanguineus

[0197] Summary

[0198] A laboratory study was conducted to determine the acaricidal properties of three compounds according to the invention (SN0305, SN0306 and SN0307) against Rhipicephalus sanguineus. Each compound was tested at 1, 10 and 100 ppm, and compared to a negative control corresponding to solvent alone (0.1% DMSO). Amitraz at 100 ppm was used as a positive control.

[0199] Methodology

[0200] Test Systems

[0201] Brown dog ticks, Rhipicephalus sanguineus, of 2.sup.nd stage, from a single cohort were used.

[0202] Tested Compounds and Application

[0203] Three (3) compounds were tested, as well as a reference compound (Amitraze). For each compound tested and Amitraz, four doses (1000, 100, 10 and 1 ppm) were applied to the ticks topically. A negative control (acetone) and an untreated group were included for comparison. The experiments were carried out in triplicate.

[0204] Experimental Protocol

[0205] Ten ticks were placed in a clear plastic container, measuring approximately 120 mm in diameter and 45 mm in height. The ticks were then anesthetized by direct exposure (for 10 seconds) to carbon dioxide. The treatments were then applied to the back (scutum) of each tick, using a Hamilton syringe, up to 0.5 μl of treatment per tick (1 application).

[0206] Cotton soaked in water was placed in each container to provide humidity throughout the experiment. The container was also vented throughout the experiment with a tube to introduce air into each container. The containers were maintained at a temperature of 23±3° C. Live, shocked and dead tick counts were taken daily after treatment for 7 days.

[0207] Statistical Analyzes

[0208] The averages in percentage of each evaluation category (healthy, knocked down and dead) were calculated, as well as the standard error. An analysis of variance (ANOVA) was performed using Minitab software, 16.sup.th edition.

[0209] Results and Conclusions

[0210] The results are shown in FIG. 14.

[0211] 1000 ppm Treatments

[0212] The treatments at 1000 ppm resulted in 100% mortality for the compounds SN0305, SN0306, SN0307 and amitraz after 7 days of treatment, with no statistically significant difference between the compounds.

[0213] 100 ppm Treatments

[0214] The 100 ppm treatments resulted in a mortality of 100%, 93.3%, 50% and 96.7% for the compounds SN0305, SN0306, SN0307 and amitraz respectively after 7 days of treatment. With a significantly higher level of mortality at 7 days for compounds SN0305, SN0306 and amitraz compared to the compound SN0307.

[0215] 10 ppm Treatments

[0216] The treatments at 10 ppm resulted in a mortality of 93.3%, 66.7%, 23.3% and 46.7% for the compounds SN0305, SN0306, SN0307 and amitraz respectively after 7 days of treatment. The compound SN0305 resulted in significantly higher mortality compared to all other treatments. The compound SN0306 caused significantly higher mortality compared to SN0307, but not compared to amitraz.

[0217] Treatments at 1 ppm

[0218] The treatments at 1 ppm resulted in much lower mortality, with 20%, 26.7%, 26.7% and 30% for the compounds SN0305, SN0306, SN0307 and amitraz respectively after 7 days of treatment.

[0219] Negative Controls

[0220] The mortality observed with the controls remained low with 3.3% mortality reported with acetone and 6.7% mortality observed in the untreated control group, after 7 days.

Example 5: Demonstration of the Acaricidal Effect in the Hive and in a Situation Outside the Brood of the Compound of Formula (I) Against Varroa destructor

[0221] Materials and Methods

[0222] Preparation of Hives

[0223] 18 hives of bees were divided into 6 groups of 3 homogeneous hives as regards the strength of the colonies (number of bees) and the Varroa infestation.

[0224] The queen of each hive was caged 3 weeks before the application of the tested compounds. The application of the tested compounds was therefore carried out in a situation outside the brood, which allows the forced passage of the Varroa mites from their reproduction phase to their phoresy phase (they are therefore all present on the bees).

[0225] The tested compounds were applied in the form of a gel three weeks after the queen was caged. The activity of the compounds was studied for a period of 9 days.

[0226] Following these 9 days of treatment, a control treatment with oxalic acid was carried out (duration of treatment equal to 1 week).

[0227] Counts of Varroa mites falling on the floor were carried out at regular time intervals to quantify the number of dead Varroa mites.

[0228] The efficiency of the treatment was calculated by taking the sum of all the dead Varroa mites from D0 to D9 (tested compounds) and dividing it by the sum of all the dead Varroa mites from D0 to D16 (tested compounds+control treatment).

[0229] The mortality of the bees was also quantified in order to demonstrate a potential toxicity of the compound tested. For this purpose, dead bee traps disposed in front of the hives were used.

[0230] Test of the Compounds

[0231] Tested Compounds

[0232] The following compounds were tested: [0233] Dexmedetomidine at 55 mg/hive in hydrochloride form, [0234] Dexmedetomidine at 110 mg/hive in hydrochloride form, [0235] Dexmedetomidine at 220 mg/hive in hydrochloride form, [0236] Detomidine at 38.4 mg/hive in hydrochloride form, [0237] Detomidine at 110 mg/hive in hydrochloride form, [0238] Detomidine at 220 mg/hive in hydrochloride form,

[0239] Results

[0240] The results concerning the calculation of the efficiency of the tested compounds are presented in FIG. 14. There clearly appears a dose effect of Dexmedetomidine on the efficiency against Varroa mites. The most effective dose was 220 mg/hive (94.5% total efficiency).

[0241] With regard to Detomidine, the results showed that the dose of 220 mg/hive allows to have a Varroacid effect.

[0242] Dexmedetomidine was therefore the most effective compound against Varroa mites.

[0243] The results concerning the mortality of the bees are presented in FIG. 15. The results showed an increase in the mortality of the bees with Detomidine at 220 mg/hive. On the other hand, no increase in bee mortality was observed with Dexmedetomidine.

[0244] These results confirm the interest of Detomidine and Dexmedetomidine as a Varroacid, in particular Dexmedetomidine which requires lower doses and is not toxic to bees.

REFERENCES

[0245] [1] Macedo, P. A., WU, J., and ELLIS, Marion D. Using inert dusts to detect and assess Varroa infestations in honey bee colonies. Journal of Apicultural Research, 2002, vol. 41, no 1-2, p. 3-7 [0246] [2] Kretzschmar A. (2016). APIMODEL, modélisation fonctionnelle de l'activité des colonies d'abeilles pour caractériser des seuils de dysfonctionnement à l'échelle du rucher. Généralisation à partir de la miellée sur lavandes. INRA-BioSP, Avignon. http://w3.avignon.inra.fr/lavandes/biosp/rapportfinal2016.pdf