RU2340181C1 - Insectoacaricide agent insektal-plus - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: insectoacaricide agent, contains permethrin, fipronil and the auxiliary substances chosen from group isopropyl alcohol, dimethyl-sulfoxide, polyethyleneglycol-400, glycerinum, surface active agent, polyvinyl pyrol etc, water, triethyl glycol. Thus the content of active substances makes: permethrin 0.2-25.0% and fipronil 2.0-15.0%. The agent can be executed in the form of a solution and is used in the form of drops on a skin of an animal, thus the permethrin content makes 1.0%, fipronil - 5.0%. The agent can be executed in the form of shampoo and the permethrin content makes from 0.2 to 15.0% and fipronil - from 2.0 to 15.0%. The agent can be executed in the form of a spray and the permethrin content makes from 1.0 to 25.0% and fipronil - from 2.0 to 15.0%. The agent can be executed in the form of a concentrate for applying on a collar the permethrin content makes 1.0 % to 15.0% and fipronil - from 2.0 to 15.0%.

EFFECT: provides stability during the long period of storage.

15 tbl, 4 ex, 5 cl

Description

The invention relates to the pharmaceutical industry and relates to the creation of tools for combating blood-sucking insects and diseases that they cause.

Arachnoentomoses of domestic and carnivores - scabies and itching mites, skin-eaters, lice-eaters, fleas, lice create many problems for animals and their owners. Scabies, itchy ticks, skin-eaters affect the skin of animals, cause itching and dermatitis. Entomotic diseases caused by insects: lice, fleas, lice, cause dermatitis and hair loss.

The leading place in the system of combating these parasites is occupied by insecticaricides. For a more effective struggle and to prevent the negative aspects of the application, scientists and practitioners are faced with the task of finding effective means in order to have a sufficient range of them. Recently, researchers have focused on the creation of insecticaricidal preparations containing synthetic pyrethroids (permethrin, cypermethrin, deltamethrin, etc.) as the main active substance (RU 2027365, 1995).

Based on this group of compounds, a significant number of preparative forms are made: shampoos, lotions, liniment, drops, etc., in which pyrethroids are used both as individual substances and in combination with other drugs. Pyrethroids easily penetrate the cuticle and respiratory apparatus, quickly act on insect nerve cells, cause agitation, convulsions, paralysis and death. Pyrethroids are practically safe for the environment and the animal organism, as they are easily hydrolyzed in the animal’s body, accumulate weakly in the environment, are not dangerous for living organisms, with the exception of fish and beneficial insects (RU 2146090, 2000).

Another well-known group of acaricides is fipronil. According to a number of studies, the drug has a wide range of insect-acaricidal action, is active against scabies mites and insects.

Carnivores, shampoos, sprays, etc. are more often used to combat flightless insects of carnivores. Recently, the stop-on method has been widely used abroad to treat sick animals with the drug and apply it to limited areas of the body.

Studies done on the development of stop-on drugs have shown that sufficient effectiveness of these forms of drugs is achieved by applying the necessary amount of the drug to a limited area of the body surface, usually in the neck, tail root, etc. Therefore, the stop-on form has an advantage over the “buy”, because full coverage of the animal with the drug is not required. A feature of stop-on - insecticides is that even when applied to a limited surface of the body, hematrophic parasites (ticks) are affected in places remote from the treated surface.

A disadvantage of the known means is the short duration of the action.

The objective of the present invention is to increase the period of insecticaricidal activity.

To solve this problem, an insect-acaricidal agent containing permethrin, fipronil and excipients selected from the group of isopropyl alcohol, dimethyl sulfoxide, polyethylene glycol-400, glycerin, surfactant, polyvinylpyrrolidone, water, triethyl glycol was proposed, while the content of active substances is 0.2-25 , 0% and fipronil 2.0-15.0%.

The tool can be made in the form of a solution and is used in the form of drops on the skin of an animal, while the content of permethrin is 1.0%, fipronil 5.0%.

The tool can be made in the form of shampoo and contains permethrin from 0.2 to 15.0% and fipronil from 2.0 to 15.0%.

The tool can be made in the form of a spray and contains permethrin from 1.0 to 25.0% and fipronil from 2.0 to 15.0%.

The tool can be made in the form of a concentrate for application to the collar and contains permethrin from 1.0% to 15.0 and fipronil from 2.0 to 15.0%.

The tool is conditionally named - "Insectal-plus."

Insectal-plus is a complex drug intended for the treatment and prevention of arachnoentomiasis in dogs and cats.

The tool can be made in the form of a spray, shampoo or concentrate for application to the collar.

Examples of the preparation of the claimed dosage forms.

ZOOSPREY

Fipronil from 2% to 15%

Permethrin from 1% to 25%

Solvents up to 100%

COLLAR

Fipronil from 2% to 15%

Permethrin from 1% to 15%

PVC up to 100%

ZOOSHAMPUN

Fipronil from 2% to 15%

Permethrin 0.2% to 15%

Surfactant + Water up to 100%

Examples illustrating the invention.

Example 1. "Drops for cats and dogs"

Pour sequentially into the tank: triethylene glycol, DMSO. Heated to 40-45 ° C. Polyvinylpyrralidone, fipronil, permethrin are poured into the solution. Stir until completely dissolved. Cool to room temperature. Dilute with isopril alcohol to the desired concentration of DV (5% - fipronil, 1% - permethrin).

Example 2. "Spray for cats and dogs"

Isopril alcohol and glycerin are poured into the tank and heated to 40-45 ° C. Fipronil and permethrin are poured into alcohol. Stir until completely dissolved (5% - fipronil, 1% - permethrin).

Example 3. "Shampoo for cats and dogs"

The required amount of water is poured into the tank. Surfactants (surfactants) are charged. The mixture is heated to 55-60 ° C. Stir until complete dissolution of the surfactant. In the resulting solution, add 5% - fipronil, 1% - permethrin. Mix to a clear color. Add to the mixture: salt, citric acid, dye, preservative. Stirred for 15-20 minutes.

Example 4. "On the collar for cats and dogs"

Cable plastic (granulate) is loaded into a heated heating mixer. An alcohol solution is prepared (10% - fipronil, 2% - permethrin). The DV solution is poured into the mixing apparatus. Stir the mixture to 60 ° C until complete alcohol evaporation. The resulting concentrate is fed to an extruder for the manufacture of a collar PVC tape.

The product is a complex preparation, which contains permethrin and fipronil as an active substance, a uniform transparent liquid with a yellowish or brownish tint with a faint specific smell.

The method of accelerated aging at elevated temperature was used to study the stability of Insectal-plus. The solutions were packaged in orange glass bottles with a capacity of 100 ml. Bottles were closed with buried polyethylene and plastic stoppers screwed on top. The solutions were stored in a thermostat at an elevated temperature of 40 ° C. The main criteria for evaluating the stability of solutions were: their appearance, pH value, quantitative content of active substances in Insectal-plus during storage. Appearance was determined visually after placing them in test tubes with a capacity of 10 cm ³ . The pH value was determined potentiometrically using a pH meter. The quantitative content of active substances (mass fraction in%) was determined for pyrethroids using gas-liquid chromatography, and fipronil was determined by HPLC.

The test results showed that the drug under this storage mode does not change its appearance within 950 hours of storage. The pH of the drugs during the entire storage period remained unchanged.

The results of the quantitative determination of drugs in solutions are presented in table 1.

Table 1
Quantitative content of DV in Insectal-plus during storage Insectal Plus Structure The initial concentration of DV,% Shelf life, hour 1320 1528 Episode 1 permethrin
fipronil 1.0 ± 0.1
5.0 ± 0.2 0.98
4.96 ± 0.2 0.96
4.95 Series 2 permethrin
fipronil 1.0 ± 0.1
5.0 ± 0.2 0.97
4.88 0.95
4.73

Thus, the results of the studies showed that both series of the drug are characterized by high stability during storage.

A study of the acaricidal activity of Insectal-plus was carried out on isolated Ps mites. cuniculi according to the "Guidelines for the initial selection of new acaricides and a comparative study of their activity against sarcoptoid ticks."

For this purpose, 6-8 concentrations of Insectal-plus were prepared using the serial dilution method, ensuring the death of ticks from 0 to 100%. Studies were performed on ticks taken on the day of the experiment. Adult mites (10 animals each) were selected with a dissecting needle on black tissue napkins 12x12 cm in size. The preparation was applied on napkins with a lower concentration of 1 ml. Napkins were folded in a knot, tied with a thread, provided with a label, folded into Petri dishes and placed in a thermostat at a temperature of 27-30 ° C at a relative humidity of 90%. The control was wetted with distilled water. With each of the concentrations of Insectal-plus, experiments were performed in triplicate. Assessment of acaricidal activity of the drug (SK-50) was carried out according to the results of the state of ticks after 24 hours. The value of SK-50 for ticks was calculated according to the Kerber formula. SK-50 of Insectal-plus for adult ticks Ps. cuniculi is 0.003%.

A study of the acaricidal efficacy of Insectal-plus in rabbit psoroptosis.

For the experiment, 9 spontaneously invaded rabbits weighing 3-3.5 kg were selected, of which three groups were formed. The drug was used once and twice with an interval of three days, 0.5 ml for each ear of the animal.

In the first group of rabbits (3 goals), the affected auricles were moistened with Insectal-plus. The second group of rabbits (3 goals) with Front Line. Three animals of the control group were treated with warm saline in the same doses on the inner surface of the auricles. The treatment was carried out without removing crusts and scabs from the affected skin.

The processing efficiency was controlled by the method of acarological examination of skin scrapings on the 5th day after the first, on the 4th day after the second treatment. On the 5th day after treatment, the following changes occurred in the first group of animals: the auricle inflammation subsided, peeling of the crusts, scabs and crusts was easily and painlessly peeled off. Microscopy revealed individual tick segments. On the fourth day after the second treatment, the inner surface of the skin of the auricles and ear canals became elastic, the crusts and scabs completely exfoliated. Acarological examination showed a complete absence of live ticks in skin scrapings.

In animals of the second group treated with Frontline, on the 5th day after the first treatment of inflammation of the skin of the auricles and auditory passages under the crusts was not visually observed. The color of the scabs became lighter, some of them were easily peeled off. Microscopic examination of scrapings from the auricles in all experimental rabbits showed no live ticks. After 4 days of the second treatment, traces of crusts and scabs on the inner surface of the ears and ear canals in rabbits of the second experimental group were not found. Acarological research showed a complete absence of live ticks in scrapings of the skin of experimental animals. During the entire observation period, there were no signs of anxiety in the treated rabbits. In control rabbits, the auricles were covered with scabs and crusts, the animals showed anxiety and scratched their ears with their paws. Microscopy observed live ticks of various degrees of development.

Study of the acaricidal efficacy of Insectal-plus against dog otodectosis

We have previously examined and selected 8 dogs of various breeds affected by otodectosis. Visually, the presence of brown crusts and scabs was observed in animals in the auricles. Acarological examination of scrapings from the inner surface of the auricles confirmed the presence of live otodectotic mites. From a microscope at a 10-fold magnification, from 4 to 9 specimens were found in one field of view.

For a comparative assessment of the therapeutic efficacy of preparations, animals were conditionally divided into 3 groups: two experimental (3 dogs each) and one control (3 dogs). The degree of damage to the ears of the experimental animals was more pronounced than in the control.

During the experiment, the effect of these drugs on the hematological parameters of the blood of animals was studied. Dogs took blood from the tarsal vein before treatment after one hour: on the 1st, 5th and 10th days. Dogs were treated twice. The interval between the first and second treatment was three days. Three dogs of the first group received Insectal-plus 5 ml in each affected ear, followed by massage of the auricle. Dogs of the second group were treated in a similar way with Front Line. Control animals were injected with 0.5 ml of heated distilled water in each damaged auricle. Processing the ears of animals was performed without removing the crusts and scabs. The effectiveness of the treatments was controlled by the method of acarological examination of skin scrapings on the 5th day after the first and 5 days after the second treatment. The results of studies of drugs showed that they have a strong acaricidal effect against ticks of O. canis. After using the preparations, ticks die, necrotic crusts and scabs soften and are rejected. Microscopic examination of the scrapings of the ears of the animals of the experimental groups 5 days after the application of live mite preparations was not found. In the control group of dogs, microscopes were observed for live ticks at different stages of development.

Repeated acarological studies of animals from experimental groups 5 days after the second treatment did not reveal live otodectotic mites in the scrapings of the auricles. In places of torn scabs and crusts, epithelization and growth of the coat were noticeable. No signs of anxiety in the treated animals with Insectal-plus were observed. Thus, complete clinical recovery of animals occurs 5 days after repeated use of drugs. The results of hematological studies of animal blood are shown in tables 2 and 3.

table 2 Hematological blood counts of dogs treated with Insectal-plus twice Duration of observation Hemoglobin, mg% White blood cells, 10 / L Erythrocytes, 10 / l ESR, mm / hour to experience 9.3 ± 0.6 7.3 ± 0.8 6.8 ± 0.9 6.1 1 hour 9.1 ± 0.4 7.8 ± 0.6 7.0 ± 0.8 5.8 1 day 10.2 ± 0.6 7.2 ± 0.8 6.2 ± 0.8 6.0 5 days 10.0 ± 0.7 7.1 ± 0.7 7.2 ± 0.9 6.3 10 days 10.0 ± 0.7 7.3 ± 0.4 6.8 ± 0.5 6.1 Note: in all groups p> 0.1

Table 3 Hematological blood counts of dogs treated with Frontline twice Duration of observation Hemoglobin, mg% White blood cells, 10 / L Erythrocytes, 10 / l ESR, mm / hour to experience 9.0 ± 0.3 6.8 ± 0.3 6.6 ± 0.7 6.2 1 hour 8.8 ± 0.6 7.3 ± 0.7 7.2 ± 0.4 6.0 1 day 9.4 ± 0.2 7.2 ± 0.8 7.0 ± 0.8 6.3 5 days 9.2 ± 0.4 7.0 ± 0.6 7.3 ± 0.6 6.4 10 days 10.0 ± 0.6 7.2 ± 0.4 7.0 ± 0.8 6.0 Note: hematological blood parameters of control animals were within the physiological norm.

As can be seen from tables 2 and 3, all indicators are within the physiological norm and do not significantly differ from the background (p> 0.05). Thus, we can conclude that the treatment of dogs with Insectal-plus does not affect the hematological parameters of blood.

Biological research

Acute toxicity

To determine the acute toxicity of the Insectal-plus preparation, the preparation was administered to mice intragastrically in the form of an aqueous emulsion, once using a syringe with olive. The toxicity of the drugs was studied on outbred white mice with a body weight of 18-20 g.

The experimental determination of the LD ₅₀ value was composed of two experiments: approximate and detailed. The first is mandatory in any case: in it, on a small number of animals, they received the necessary preliminary information in the dose range close to the average. In a detailed experiment, 4 different doses were tested (each for at least 6 animals), provided that the extreme doses act as LD ₀ -LD ₁₀₀ .

Experimental animals were monitored for 7 days. During this period, attention was paid to the clinical condition and behavior, the reaction to external stimuli, the possible dynamics of the development of intoxication and the death of animals. The parameters of acute toxicity (LD value ₅₀ ) were calculated according to the method of G. F. Lakin, which is based on taking into account the mortality of animals from the administered doses. The level of toxicity of Insectal-plus was characterized according to GOST 12.1.007.76 “Harmful substances, classification, safety requirements”. In white mice, when toxic doses of the drug were injected into the stomach, an oppressed state was observed, rapid breathing, the hair was disheveled, a dull shade prevailed.

When lethal doses of the drug were administered to white mice, they were immediately depressed: the animals were inactive, did not respond to external stimuli, and died within 24 hours after using the drug.

A pathological and morphological study of dead animals revealed: hemorrhages on the epicardium of the heart bag, pulmonary edema, degenerative changes in the liver and kidneys.

Studies have shown that according to G. F. Lakin, the average lethal dose of LD ₅₀ of Insectal-plus was 543 mg / kg body weight. According to the classification of GOST 12.1.007.76, the drug belongs to moderately hazardous substances - hazard class 3.

The study of the annoying properties of the drug Insectal-plus

1. A study of the local action on the mucous membrane (eye test).

Six rabbits acted on the conjunctiva according to the method of V.V. Gatsur (1974). Clinically healthy rabbits that were quarantined for seven days were kept on a standard diet. In the conjunctival sac of the right eye, three drops of the first group of rabbits were pipetted with one drop of Insectal-plus, the left eye served as a control, 2 drops of distilled water were introduced into it. Evaluation of the irritant effect was carried out after 1, 4, 6, 12 and 24 hours visually by changing the conjunctival blood flow, the presence of lacrimation and the state of the cornea in a 10-point system according to Scheme 1.

Scheme 1 Reaction rate Scoring Annoying effect lack of reaction 0 absent weak reaction 2 weak severe hyperemia four mild the presence of lacrimation 6 moderate the presence of discharge 8 expressed prolonged, pronounced hyperemia, lacrimation, swelling of the eyelids 10 pronounced

Table 4 Assessment of the irritating effect of Insectal-plus when exposed to the rabbit eye conjunctiva (3 animals per group) Group Injected drug Score in points (hours) Annoying effect one four 6 12 24 one Insectal Plus four four 6 2 0 Weak
expressed
moderate four four 6 2 0 four four 6 2 0 2 water 0 0 0 0 0 absent 0 0 0 0 0 0 0 0 0 0

When Insectal-Plus was introduced into the conjunctive sac of rabbits, transient hyperemia and lacrimation were noted. These phenomena completely disappeared after 24 hours, which indicates the expression of the effect of the drug on the mucous membrane.

2. The study of the local action of the drug Insectal-plus on the skin, coat and hematological parameters of the blood of rabbits (the number of leukocytes, red blood cells, hemoglobin level, ESR).

A study of the irritant effect of Insectal-plus on the skin was carried out on 12 white rabbits weighing 3-3.5 kg (4 animals in the experimental groups and 4 in the control group). To conduct the experiment, two sections of 5x3 cm were cut out on the back in the direction of the hair to the tail. Then, 4 rabbits of the first group were injected daily and rubbed 1.5 ml of Insectal-plus into the hair and skin for 5 days. 4 other animals of the second group were infused with Frontline using the same technique. Control animals were rubbed with physiological saline using the above procedure. A visual assessment was carried out daily, taking into account the behavior of animals and the condition of the skin and coat at the rubbing sites.

In rabbits, blood was taken from the ear marginal vein: before treatment, after 1 hour, on 1, 5, 10 days after treatments.

Table 5 Hematological blood counts of rabbits treated with Insectal-plus Duration of observation Hemoglobin, mg% White blood cells, 10 / L Erythrocytes, 10 / l ESR, mm / hour Before experience 12.3 ± 0.2 6.8 ± 0.7 7.5 ± 0.4 4.0 ± 0.5 1 hour 11.8 ± 0.6 6.5 ± 0.8 7.2 ± 0.6 3.6 ± 0.4 1 day 12.2 ± 0.3 7.1 ± 0.4 7.3 ± 0.4 4.0 ± 0.6 5 days 12.4 ± 0.3 6.4 ± 0.7 7.3 ± 0.5 3.4 ± 0.3 10 days 11.8 ± 0.6 6.9 ± 0.4 7.4 ± 0.7 3.8 ± 0.8

Table 6 Hematological blood counts of rabbits treated with Frontline Duration of observation Hemoglobin, mg% White blood cells, 10 / L Erythrocytes, 10 / l ESR, mm / hour Before experience 10.8 ± 0.5 6.7 ± 0.4 7.3 ± 0.5 3.2 ± 0.4 1 hour 11.2 ± 0.3 6.4 ± 0.6 7.2 ± 0.2 3.2 ± 0.3 1 day 11.4 ± 0.4 6.2 ± 0.6 7.3 ± 0.4 4.0 ± 0.5 5 days 11.6 ± 0.3 6.8 ± 0.5 7.0 ± 0.6 3.8 ± 0.6 10 days 12.2 ± 0.2 6.5 ± 0.7 7.2 ± 0.4 3.6 ± 0.4

Studies have shown that a 5-fold rubbing of drugs in the specified dose does not cause changes in the behavior of animals, does not lead to their death, reddening of the skin, scratching, swelling, thickening of the skin fold, and discoloration of the coat are not observed. There was no painful reaction on palpation. Studies of hematological parameters (Tables 5 and 6) indicate that Insectal-plus and Frontline preparations are low-toxic substances that do not cause any morphological changes in the blood, even when applied repeatedly to the skin of animals.

3. Allergy properties of Insectal-plus

The experiments were conducted on 12 guinea pigs using histamine shock according to the method of M. L. Gershkovich (1954).

Table 7 Allergic properties of the drug Insectal-plus Group amount
animals A drug,
ml Time
introducing
histamine
in an hour Time
offensive
death
animals from
histamine
shock, min Death
animals
% one four
four Insectal
a plus 1,5
3 17, ± 0.4
17.4 ± 0.5 one hundred
75 The control four one hundred

As studies have shown (table 7), the drug Insectal-plus, administered to guinea pigs subcutaneously at a dose of 2 ml / kg, did not prevent the death of animals from histamine shock, but did not accelerate it compared with the control. The reaction in the control and experimental animals to the introduction of histamine was the same and was manifested by anxiety, increased breathing, taking a lateral position of the body, convulsions and ended in death.

The significance of differences in all cases was less than 0.05, i.e. the time of death of the experimental animals did not significantly differ from the indicators of the experimental groups; therefore, the Insectal-plus preparation at these concentrations does not have an allergic effect.

The effect of Insectal-plus on the antitoxic function of the liver.

The study of the effect of drugs on the antitoxic function of the liver was carried out according to the method proposed by E. L. Shchedrin et al. (1988).

4 groups of mice were formed, 20 animals each with an initial body weight of 19-20 g. Animals of 2 experimental groups received the test drug daily in increasing doses from 0.045 ml / kg to 0.344 ml / kg for 24 days. Animals of the control group received nothing. After 24 days, experimental and control mice were injected subcutaneously with 0.2 cm sodium nitrate at a dose of LD ₅₀ equal to 350 mg / kg body weight with an interval of 30 seconds. Subsequently, the time of occurrence of the death of animals was recorded. From each group, 2 determinations of the least and longest period of action of the toxin on the animal organism were excluded and the average time of death of the mice in the groups was calculated.

As our studies showed, the animals of the control group died after 20 minutes, the animals of the experimental groups after 19 minutes and 40 seconds, i.e. the time of death of animals is the same, even the instantaneous administration of drugs to control animals does not affect the antitoxic function of the liver.

The study of the efficacy, tolerability and safety of the drug Insectal-plus in dogs affected by trichodectosis.

Studies on the effectiveness, tolerability and safety of the drug were conducted on 8 dogs of the previous experience. When examining the outer integument of dogs on the head, on the root of the tail, on the inner surface of the paws, we found insects. In an area of 10 cm ^{2, the} susceptibility ranged from 3 to 9 specimens. The type of causative agent of dog parasites is the beetle Trichodectes canis.

Dogs were divided into 2 groups A and B (4 dogs each). In group A for 3 dogs, Insectal-plus was used in doses of 1, 2, 3 ml. The preparation was applied by the stop-on method to the nape of the dogs and to the root of the tail.

In group B, Frontline in doses was used in the same number of animals according to the instructions for use.

Two dogs (one from each group) were not treated with the preparations and kept together with the treated animals.

In group A: 1 ml of Insectal-plus was consumed on the first dog (weight 9 kg), 2 ml on the second (weight 18 kg), and 3 ml on the third (weight 20 kg).

The animals were monitored daily for the first 5 days. For the condition of the skin and coat - every other day. Acrological studies were performed on the 5th and 10th day after treatment. The application of Insectal-plus and Frontline preparations to the coat in test doses caused a noticeable interest in all animals in the first quarter of an hour. Two dogs from group A and one from group B had an attempt to lick the treated area of the body in the back. Subsequently, the animals behaved calmly. Visual examination and palpation in the field of application of the preparations to the skin in none of the groups revealed its changes after 24 hours. Hyperemia, swelling, and peeling of the skin in all animals were not detected on the 5th day. The results of an acarological study showed that in a dog treated with Insectal-plus at a dose of 1 ml per animal, the number of lice-eaters decreased in all areas of the coat. On the area of 10 cm ^{2 the} lesion was 1-3 copies. In the second and third dogs treated with Insectal-plus in a volume of 2 and 3 ml per animal, only live and dead insects were found in small quantities 1.5 hours after application of the drug in the head area, in two sections of 5x8 cm in size.

Treatment of Group B animals with Front Line showed that watering the test dogs significantly reduces the number of insects on the animal’s hair, but does not completely destroy them. Only repeated treatment of animals with Insectal-plus and Frontline preparations showed that drugs in doses of 1-3 ml show 100% effectiveness against trichodectosis of dogs. Consequently, under the influence of Insectal-plus and Frontline preparations, adults and larvae perish during a single treatment, but viable eggs remain. Therefore, animals should be treated with Insectal-plus and Frontline with an interval of 7-10 days.

A visual examination of the skin did not reveal scratching, thickening, the possible formation of dermatitis, seborrhea, hair loss, and skin dying in animals.

Insectal-plus was tested by practicing veterinarians with otodectosis and notothedrosis of cats, trichodectosis of dogs (vlosoedov), siphonapterosis of dogs and cats (losiness), menognatosis (lice) of dogs.

Studies conducted in laboratory and industrial conditions to study the toxicological, acaricidal and biological properties of the developed drug, give reason to recommend it for the fight against sarcoptosis and entomoses of domestic and carnivorous animals.

In addition, the following tests were carried out.

The study of acaricidal efficacy of the drug on ticks Ps.cuniculi

The acaricidal activity of the drug was studied on P.cuniculi ticks according to the Guidelines for the initial selection of new acaricides (1982).

Ticks were selected on the day of the study. For each concentration of the studied drug, 10 adult mites were selected on black napkins, which were wetted with the drug emulsion (1 ml) with different content of DV. Then the ticks were placed in a thermostat at a temperature of 30 ° C and a relative humidity of 90% and their condition was evaluated 1, 4, 6, 24, 48, and 72 hours after application of various concentrations of the drug. The experiment was carried out in triplicate. The value of SK _{50 was} calculated according to the results of experiments after 24 hours. As a result of the work carried out, it was found that the SC ₅₀ value of the drug was 0.028%.

Table 8 Acaricidal activity of Insectal-plus in an acute experiment with Psoroptes cuniculi Indicators The concentration of the drug,% 0.1 0.08 0.06 0.04 0.02 0.01 0.005 0.01 Survived - 2 four eleven 13 17 22 thirty Died thirty 28 26 19 17 13 8 - Z 29th 26 22.5 eighteen fifteen 10.5 d 0.02 0.02 0.02 0.02 0.01 0.005 Zd 0.58 0.52 0.45 0.36 0.15 0,0525 Note: Z is half the total number of insects that have fallen from the last two doses;
d is the difference in the magnitude of subsequent doses;
n = 30
CK ₅₀ = 0.1 - 2.11 / 30 = 0.028% by DW

The residual acaricidal effect of the drug in the laboratory was studied using the method of forced contact of Ps.cuniculi mites for 15 minutes with surfaces treated with an aqueous emulsion of the drug at a flow rate of 1 ml per 10 cm ² . Contacting was carried out after 5, 7, 10, 15, 20 and 25 days after treatment. The results are presented in table 9.

As a result of experiments on the study of the duration of residual acaricidal action, it was found that the drug has a long residual effect: by contacting ticks Ps. cuniculi with wooden surfaces treated with 0.1% aqueous emulsion, 100% death of ticks was observed within 15 days, with treated 0.025% and 0.05% during 7 and 10 days, respectively.

Table 9 Duration of residual acaricidal action of Insectal-plus in experiments on P. cuniculi The concentration of the drug (%) Qty
mite experiences Tick mortality (%) after (days) 5 7 10 fifteen twenty 25 thirty 0,025 3 90 one hundred one hundred 72 54 eighteen 0 0 0.05 3 90 one hundred one hundred one hundred 81 54 eighteen 0 0.1 3 90 one hundred one hundred one hundred one hundred 72 eighteen 5.5 water 3 90 2 2 one one 0 0 0

The insecticidal activity of the preparation was investigated by the standard method of planting insects on the surface treated with the preparation on the insect culture of Ctenophalides canis fleas.

Surfaces 10x10 cm in size from unpainted plywood were treated with a preparation with a flow rate of 50 ml / m ² . The experiments were carried out on three surfaces using 10 insects in each series. The contact time with the treated surface was 5 and 10 minutes. After this, the fleas were transferred to clean vessels. Accounting for the death of insects was carried out after 0.5; one; 3 and 24 hours.

The control was surfaces treated with the drug without permethrin and water. The research results are presented in table 10.

Table 10 Insecticidal activity of Insectal-plus in an acute experiment with Ctenophalides canis Drug concentration Consumption rate, ml / m ² Exposure, min The death (%) of insects, through 0.5 h 1.0 h 2.0 h 24 h 0.5 fifty 5 one hundred one hundred one hundred one hundred 0.05 fifty 10 one hundred one hundred one hundred one hundred 0.05 fifty 5 one hundred one hundred one hundred one hundred Without DV fifty 10 16.6 16.6 twenty twenty water fifty 10 0 3.3 6.6 6.6

The results showed that the emulsion of the drug with a content of 0.5% and 0.05% DV has a pronounced insecticidal activity upon contact with the treated surface for 10 and 5 minutes, at a flow rate of 50 ml / m ² .

In the control (the drug without permethrin and fipronil and water), insect death was 20% and 6.6%, respectively.

In order to determine the duration of the residual effect of the drug, plywood plates were treated with a drug with a content of 0.05% DV with a flow rate of 50 and 100 ml / m ² and then insects were contacted with it using a Nabokov exposure meter for 15 minutes. The experiments were carried out on three surfaces, using 10 fleas in each experiment. Flea contacting was performed 3, 5, 10, 20, and 25 days after surface treatment. After contacting the insects, they were placed in test tubes and their death was recorded. The results are presented in table 11.

Table 11 Duration of insecticidal action of Insectal-plus on surfaces Consumption rate, The death of fleas (%) upon contact in (days) after treatment 0.05% DW, ml / m ² 3 5 10 twenty 25 thirty fifty one hundred one hundred one hundred 96.6 90 fifty one hundred one hundred one hundred one hundred 96.6 93.3 53.3

As the results of studies showed, the drug at 0.05% concentration by DW has a long residual effect on the surfaces. The death of 100-90% of insects is noted within 25 days after treatment.

The study of insectacaricidal efficacy of Insectal-plus in animal experiments

The tests were conducted at the clinic "Junior" (Moscow). The object of the study was dogs and cats of various breeds, including short-haired and long-haired, affected by fleas Ctenophalides canis and lice-eating Trichodectes canis.

The intensity of animal infestation was determined by visual counting of insects on 3-4 parts of the animal’s body (tail root, inner thighs, abdomen) with an area of 100 cm ² . The preparation was applied at the rate of 0.1 ml / kg of mass onto the skin in a place inaccessible for licking (at the base of the skull, between the shoulder blades) in the control insecticidal zoo shampoo with permethrin “Chistyulya P” was used. Zoo shampoo was applied to moistened animal hair at the rate of 1.0 ml / kg and lightly rubbed into the skin with massage movements. After 5 minutes, the shampoo was washed off with warm water (25-30 ° C) and the animals were wiped with a towel. Then the hair was combed with a comb.

With the consent of the owners of the animals, 6 dogs and 4 cats were processed. The number of lice eaters before the experiment on dogs was 15–25 ind. per 100 cm ² , the number of fleas on cats is 10-20 specimens. per 100 cm ² , when examining the animals 48 hours after treatment with Insectal-plus and live shampoo live fleas, larvae and adult lice-eaters were not found.

When examining after 7 days on animals treated with Insectal-plus, no live insects were found, while on animals treated with zooshampoo, single larvae of larvae (1-3 individuals) were found, and therefore the treatment was repeated.

As a result of further observation within 30 days of the treated animals, parasites were not found.

Table 12 Insecticidal efficacy of Insectal-plus in experiments on dogs and cats Animal information Date of inspection Insects detected date of processing Insectal Plus Murka cat, 3 years old, Siberian 08/20/03
08/30/03 30 fleas
Not found 08/20/03
Not processed Kitten Marius, 6 months old, purebred, smooth-haired. 08/05/03
08/15/03 20 fleas
Not found 08/05/03
Not processed Pavka's dog, 4 months old, bezpor., Long. 06/20/03
06/27/03 25 beetles
Not found 06/20/03
Not processed Jerry dog, 10 months old, poodle 06/20/03
07/01/03 25 beetles
Not found 06/20/03
Not processed Dog Mars, 2 years old, German. shepherd dog 06/20/03
06/29/03 18 fleas
Not found 06/20/03
Not processed Dog Bulka, 1 year old, poodle 06/20/03
07/01/03 20 whipworms
Not found 06/20/03
Not processed ZOOSHAMPUN Chistyulya P Dog Paw, 7 months old, afghan 08/20/03
08/27/03
09/26/03 20 whipworms
4 larvae
Not found 08/20/03
08/27/03
Not processed Dog Chuk, 12 months old, b / n, long-haired. 06/20/03
06/27/03
07/06/03 20 whipworms
3 larvae
Not found 06/20/03
07/27/03
Not processed Cat Dusya, 2 years old, Persian 06/20/03
06/26/03 30 fleas
Not found 06/20/03
Not processed Dora cat, 8 months old, b / n, smooth-haired. 06/20/03
06/26/03 40 fleas
Not found 06/20/03
Not processed

Tests have shown that the proposed drug Insectal-plus has a pronounced insecticidal activity against insects that parasitize pets, does not cause skin irritation, dermatitis, seborrhea, allergic and other undesirable effects. Insectal-plus is convenient to use and has a longer residual effect than insecticidal shampoos containing permethrin, which ensures the destruction of hatched larvae hatching from eggs.

The study of the therapeutic efficacy of Insectal-plus in dogs sarcoptosis

In the experiments, 7 dogs of different breeds (German Shepherd, Boxer, Airedale Terrier, Bobtail, Dobermann, Briard, Fox Terrier) affected by sarcoptosis were used.

Clinical examination of dogs on the affected surfaces showed dried escudate in the form of scab deposits, flushing and skin tissue infiltration. The diagnosis of the disease was confirmed by acarological studies of scrapings and the detection of S. canis. The drug was applied to the skin at the withers along the spine at a dose of 0.1 ml / kg of body weight. Treatment efficacy was monitored by scraping microscopy. After 2 days, dead ticks, deformed larvae and eggs were found in scrapings. Clinical signs of the disease began to pass 3-4 days after the first application of the drug, which was manifested by the disappearance of hyperemia, a decrease in exudation, and the disappearance of crusts. 7-10 days after the first use of the drug, all animals recovered, which was confirmed by clinical and acarological studies. Relapse of the disease when observing animals for a month was not observed.

Table 13 Therapeutic efficacy of Insectal-plus in dogs sarcoptosis Dog breed floor age Recovery period Relapse fox terrier cob. 3 years 7 days no boxer cob. 1 year 9 days no bobtail bitch 9 months 8 days no doberman cob. 3 years 10 days no briard bitch 1 year 8 days no German Shepherd cob. 8 month 9 days no Airedale bitch 2 years 9 days no

When using the drug in the indicated doses and treatment modes in animals, no side effects and complications (oppression, refusal of feed, allergic reactions) were noted.

A production test of the proposed drug was carried out in the clinic "Junior". A total of 16 dogs with sarcoptosis were treated.

Before treatment, sick animals observed inflammation and infiltration of the affected areas. Acarological studies in scrapings revealed ticks of S. canis at different stages of development (5-9 individuals in the field of view). The treatment with the use of the drug "Irbis" in a dose of 1.0 ml / kg of animal weight provided 10 animals after a single application. In 6 animals, when examining scrapings, 7 days after the first treatment, larvae and mite eggs were found (1-3 in the field of view), and therefore, repeated treatment was performed. When examining scrapings after 5 days, ticks were not found. There were no recurrences of sarcoptosis when observing animals within 30 days.

The study of the therapeutic efficacy of Insectal-plus in cat nohedrosis

In the experiments, 6 cats affected by moderate nothedrosis (lesions in the head area) were used. The drug at a dose of 0.2 ml / kg and 0.5 ml / kg of mass was applied to the skin along the spine, drip, once or twice with an interval of 7 days. The effectiveness of the drug was determined by the presence and condition of ticks in the experiment and in the control before and after each treatment by microscopy of scrapings. After 2 days after the first treatment at a dose of 0.2 mg / kg, only dead ticks and eggs were found in cats in scrapings, after 4 days the crusts dried out, sprinkled, the skin turned pale pink, dry after repeated treatment, the skin became dry, white colors, slightly rough. Recovery of animals occurred on the 7-8th day. An acarological study of ticks, larvae and eggs was not found.

Table 14 Therapeutic efficacy of Insectal-plus in cat nohedrosis breed age recovery period relapse outbred 1,5 years 8 days no outbred 10 months 7 days no outbred 2 years 8 days no outbred 1 year 8 days no Siamese 6 months 8 days no the control outbred 14 months ticks found ticks found

The results of a study of the insecto-acaricidal efficacy of Insectal-plus indicates that

- the drug in experiments in vitro has a pronounced insecticidal activity against fleas Ctenophalides canis and acaricidal activity against sarcoptoid ticks Psoroptes cuniculi;

- the drug has a long residual effect on the treated surfaces. The death of 100-90% of insects and ticks is observed within 10-20 days after application of the drug;

- the drug at a dose of 0.2 ml / g of animal mass has high insecticidal and acaricidal efficacy in arachnoentomiasis of dogs and cats;

- the drug in therapeutic and 3-fold therapeutic doses does not cause dermatitis, side effects and complications in animals.

Insectal-plus has a pronounced insectoacaricidal activity against the larval and mature phases of the development of fleas, lice, lice beetles, ixodid and sarcoptoid ticks that parasitize dogs and cats.

After cutaneous application, the agent accumulates in the epidermis, hair follicles and sebaceous glands of the animal’s body and has a long contact insectoacaricidal effect.

The mechanism of action of permethrin, which is part of the honeycomb, is to block the transmission of nerve impulses, which causes impaired coordination of movements, paralysis and death of ectoparasites. Fipronil disrupts the passage of chlorine ions in the GABA-dependent ectoparasite receptors, the transmission of nerve impulses and the activity of the nervous system, causing the death of arthropods.

Insectal-plus, according to the degree of exposure to the body, refers to moderately hazardous substances (hazard class 3 according to GOST 12.1.007-76), when applied to the skin in recommended doses, it does not exert a locally irritating and resorptive toxic effect, if it comes into contact with the eyes, it causes irritation.

Insectal-plus is administered to dogs and cats with entomoses, sarcoptosis, notothedrosis and ixodid ticks.

The tool, spreading the hair, is applied to the skin between the shoulder blades or in the neck at the base of the skull in the doses indicated in the table:

Table 15 type and weight of the animal dose of the drug, ml cats and dogs from 2 to 10 kg 1,0 dogs from 10 to 20 kg 2.0 dogs from 20 to 30 kg 3.0 dogs over 30 kg 5,0

In order to prevent re-infestation with fleas, the animals are replaced or treated with an insecticide in accordance with the instructions for its use.

To destroy ixodid ticks on the body of an animal, a drug in the amount of 1 drop is applied to the tick and the place of its attachment to the skin. If the mite does not fall off spontaneously within 20 minutes, it is gently pulled out of the skin with tweezers and destroyed.

Repeated treatment of animals is done according to indications, not more than 1 time per month.

Patients with infectious diseases and recovering animals, pregnant and lactating females, puppies and kittens younger than 2 months of age, as well as dogs weighing less than 2.0 kg, should not be treated.

The animal should not be washed, ironed and kept out of reach of small children within 48 hours after treatment with the drug.

With increased individual sensitivity of animals to the active components of the drug and the appearance of complications (lacrimation, excessive salivation, muscle tremors, vomiting, signs of skin irritation), the drug should be washed with soap and water.

Treatment of animals with Insectal-plus should be carried out using rubber gloves.

While working with the drug is not allowed to smoke, drink and eat.

At the end of the work, wash your hands with warm water and soap, wash and dry the gloves.

In case of accidental contact with the skin and mucous membranes, rinse it off immediately with a stream of water.

It is forbidden to use empty tubes-pipettes from under the preparation for domestic purposes, they are placed in a plastic bag and disposed of with household waste.

Insectal-plus should be stored out of the reach of children.

The product is available in dosage form in the form of a spray, shampoo, etc. It has a longer duration than the existing ones (for each lex form) with either permethrin or fipronil.

Claims (5)

1. Insectoacaricidal agent containing permethrin, characterized in that it additionally contains fipronil and excipients selected from the group of isopropyl alcohol, dimethyl sulfoxide, polyethylene glycol-400, glycerin, surfactant, polyvinylpyrrolidone, water, triethyl glycol, while the content of active substances is permethrin 0 2-25.0% and fipronil 2.0-15.0%. 2. The tool according to claim 1, characterized in that it is made in the form of a solution and is used in the form of drops on the skin of an animal, while the content of permethrin is 1.0%, fipronil 5.0%. 3. The tool according to claim 1, characterized in that it is made in the form of shampoo and contains permethrin from 0.2 to 15.0% and fipronil from 2.0 to 15.0%. 4. The tool according to claim 1, characterized in that it is made in the form of a spray and contains permethrin from 1.0 to 25.0% and fipronil from 2.0 to 15.0%. 5. The tool according to claim 1, characterized in that it is made in the form of a concentrate for application to the collar and contains permethrin from 1.0 to 15.0% and fipronil from 2.0 to 15.0%.