M. B. Musaev’s research while affiliated with Federal State Institution "Scientific Center of Expertise of Medical Products" and other places

The purpose of the research is to develop the production of ivermectin-based complex solid dispersion preparations with water-soluble polymer fillers using the technique previously developed in the VNIIP – FSC VIEV with a solvent, as well as the INEOS RAS (Nesmeyanov Institute of Organoelement Compounds RAS) technique in one stage without involving liquid phases, and their comparative performance testing against parasitosis of animals. Materials and methods. Preparation 1 was developed using the technique developed in the VNIIP – FSC VIEV in several stages, and preparation 2 was developed using the INEOS RAS technique in one stage. A comparative performance test of the above preparations was conducted on three private Chechen Republic farms on 870 Tushino sheep and 57 local goats spontaneously infected with gastrointestinal Strongylata. In the first experiment, the sheep (n = 130) were divided into 4 groups of 30–35 animals each on the analog principle. The sheep from the first and second experimental groups were given preparations 1 and 2 orally in the suspension formulation at doses of 0.2 mg/kg by active substance. The third and fourth groups of 35 sheep each were treated with the same preparations mixed with combined feed at the same dose, by the group method only. In the second experiment, 140 sheep and 57 goats were treated once, individually, with preparation 1 at a dose of 0.2 mg/kg in the suspension formulation. The third experiment on testing preparation 1 was conducted on 600 sheep, 217 bull calves, 165 horses, and 15 dogs in production conditions. Results and discussion. The solubility of preparation 1 increased by 7.1, and preparation 2 by 4.7. A HPLC analysis confirmed 95.0% preservation of ivermectin in both preparations. One hundred percent efficacy against gastrointestinal nematodes in sheep was confirmed for preparations 1 and 2 obtained by different techniques, given individually, once, at a dose of 0.2 mg/kg in the suspension formulation. The treating of the sheep with preparation 1 mixed with combined feed by group feeding obtained 100% efficacy, and 94.6% for preparation 2. The deworming of the sheep in the second experiment showed 100% efficiency for preparation 1 given individually, once, at a dose of 0.2 mg/kg by active substance in the suspension formulation. The testing of complex preparation 1 at a dose of 0.2 mg/kg in the third experiment obtained 100% efficacy against gastrointestinal nematodes in horses, young cattle, sheep, goats, and dogs. No side effects were observed in the animals after deworming.

The purpose of the research is to assess the acute toxicity in laboratory animals of experimental samples of two antiparasitic combined drugs from the group of avermectins and pyrethroids, conventionally designated as CA-17 and CA-18 (NBC Pharmbiomed LLC). Materials and methods . The present experiment was performed on sexually mature outbred rats and mice (males), to which the studied drugs were administered in two ways, intragastrically and subcutaneously, in a dose range that allows calculating the parameters of acute toxic effects or setting the maximum tolerated dose. The studied preparations were transparent oily liquids containing avermectin C and cyfluthrin in different concentrations. During 14 days, the condition and behavior of the animals were monitored, death and symptoms of intoxication were recorded. Probit analysis was used to calculate LD50 values and other parameters of acute toxic effects of drugs. Results and discussion . The ЛД50 value of CA-17 when administered intragastrically to rats and mice is 6072 (5208 6975) and 4008 (2340 6094) mg/kg, respectively. The ЛД50 value of CA-18 when administered intragastrically to rats and mice is 3716 (3103 4496) and 2929 (1454 3603) mg/kg, respectively. Compared with rats, mice have a species sensitivity to both drugs. Preparations CA-17 and CA-18 for intragastric administration according to the hygienic classification belong to hazard class 3. The ЛД50 value of CA-17 after cutaneous administration to rats and mice is more than 7,500 mg/kg, respectively. Similar indicators for CA-18 for rats and mice are more than 5,000 and 2,500 mg/kg, respectively. According to the parameters of cutaneous toxicity, both drugs belong to hazard class 4.

The purpose of the research is to evaluate embryotoxic and teratogenic effects of the supramolecular complex of ivermectin aniverm-2.0%. Materials and methods . The experiment was conducted on 100 female rats weighing 180–200 g that were kept in the vivarium of the VNIIP – FSC VIEV. The experimental rats were divided into groups of 8 animals each. The supramolecular complex of ivermectin aniverm-2.0% was administered to inbred pregnant female rats orally in the suspension formulation using an intragastric catheter three times in increased doses. The 1st group pregnant females were administered a total dose of aniverm-2.0% for the first 6 days which was 3 times higher than the therapeutic dose (3.6 mg/kg) by the active substance, and 180 mg/kg by the drug. The 2nd group animals were given the ivermectin substance in a similar dose of 3.6 mg/kg in the suspension formulation on starch paste on the first day of pregnancy. The 1st group rats were given a total dose of aniverm-2.0% increased to 3-fold, equal to 7 days (7–13 days), at a dose of 4.2 mg/kg by the active ingredient, and 210 mg/kg by the drug, and the 2nd group rats received a similar dose of the ivermectin substance of 4.2 mg/kg in the suspension formulation on starch paste on day 7 of pregnancy. On day 14 (days 14–19), the 1st group rats were administered a total dose of aniverm-2.0% for 6 days for the third time at a dose of 3.6 mg/kg by the active ingredient, and 180 mg/kg by the drug, and the second group of animals received the substance of ivermectin at a dose of 3.6 mg/kg by the active ingredient in the suspension formulation on starch paste. The female rats from the 3rd control group were administered starch paste of 5–7 mL three times during the same periods of pregnancy as the experimental animals. On day 20 of embryogenesis, all experimental rats were euthanized. In the females, after slaughter and extirpation of the uterus, the number of embryos, resorptions, and corpus luteum in pregnancy in both ovaries was recorded; the craniocaudal embryo dimensions were measured, their weight was recorded, and their sex was determined. The placenta was measured and weighed to calculate the fetoplacental coefficient. Attention was also paid to deformities and developmental anomalies in the embryos. Some of the fetuses were then placed in Bouin's fluid for further examination of the internal organs using the Wilson method. Another part was fixed in 96.0% alcohol to examine the skeletal system and measure bone length using the Dawson method. To assess embryotoxic effects of the drug, preand post-implantation death and total embryonic mortality were recorded. The obtained results were processed statistically. Results and discussion . In the entire pregnancy period, no signs of intoxication were observed in the animals. The animals were active, ate food readily and gained weight well. Macroscopic examination of the internal organs did not reveal any pathological changes. There were no signs of inflammation, deformities or other abnormalities in the embryos and placentas. Aniverm-2.0% in a three-fold increased dose of 0.6 mg/kg with intragastric administration did not exhibit embryotoxic or teratogenic effects in critical periods of embryogenesis.

The purpose of the research is to test the supramolecular complex of ivermectin Aniverm-2.0% by free feeding of the drug mixed with concentrated feed against gastro-intestinal strongylatosis of the deer. Materials and methods . The test of the supramolecular complex of ivermectin Aniverm-2.0% against gastro-intestinal strongylatosis of semi-wild animals was carried out from November 2023 to April 2024 in the reserve of the country residence of the head of the Chechen Republic, where more than 200 marals, sika deer and roe deer were kept. 40 fresh fecal samples were collected from the ground in December 2023. The fecal samples were examined in the biology laboratory of the Ibragimov Integrated Research Institute of the Russian Academy of Sciences by the Fülleborn flotation method using a saturated sodium chloride solution and the method of successive washes to detect elaphostrongyl larvae. Aniverm-2.0% was fed at a dose of 0.3 mg/kg mixed with concentrated feed by the group method; evenly poured into 4 feeders for 200 deer. The efficacy of the drug was taken into account by the "critical test" after 10 days and 4 months after deworming. Results and discussion . In the feces of deer, strongylata eggs were found in 23 out of 40 samples (EI = 57.5%) with an average of 88.9±2.74 egg sp. detected in 1 g of feces. Elaphostrongyl larvae were not detected. Dicrocoelium spp. eggs were found in 8 samples. After deworming of marals, red deer and roe deer with Aniverm-2.0% at a dose of 0.3 mg/kg by active ingredient (0.15 mg/kg by the drug) by the group method mixed with compound feed, strongylata eggs of the digestive tract were not found in the feces after 10 days and for 4 months after treatment. 100% therapeutic and prophylactic efficacy of the drug was established. The animals willingly ate the drug with food, no side effects were noted after administration of the drug.

The purpose of the research is to obtain and test a complex drug Aniverm-2.0% based on the ivermectin substance by free feeding in a mixture with concentrated feed against parasitosis of cattle, sheep, and horses. Materials and methods . Aniverm-2.0% obtained by mechanochemical technology was tested on a farm in the Shatoy Region, the Chechen Republic, on 900 sheep, 180 cattle and 156 horses in March-April 2023. To determine the infection rate, coproovoscopic examinations were conducted on 80 samples from sheep, 20 samples from young cattle and 30 samples from horses using the Fulleborn’s flotation method with a saturated sodium chloride solution. Sheep were dewormed using a group method. The drug was mixed with feed per 0.3 mg/kg for the AS or 60 mg for the drug per animal. The young cattle were kept by tie-up housing and therefore they were given Aniverm-2.0% once at a dose of 0.2 mg/kg for the AS individually in a mixture with feed. The horses were given the drug in a group method at a dose of 0.2 mg/kg for the AS or 1.5 mg/kg for the drug. Fecal samples from all animals were examined before and 2 weeks after the drug. Results and discussion . A 100% rate of infection of the sheep by gastrointestinal nematodes (Strongylata, Nematodirus spp., Trichocephalus spp.) was determined with an average number of eggs 633.3 in 1 g of feces. In 20 fecal samples from the young cattle, 16 were found to have Strongylata eggs (prevalence = 80.0%) with 222.2–280.5 specimens of Strongylata eggs found in 1 g of feces. Strongylata eggs were found in all 30 examined fecal samples from the horses, on average, 253.3 specimens/g of feces (100%). At 2 weeks after treatment, nematode eggs were found in three out of 80 examined fecal samples from the sheep, which can be explained that the sheep did not eat the appropriate dose of the drug. The drug efficacy was determined to be 99.7%. One hundred percent efficacy of the drug was obtained on young cattle and horses.

The purpose of the research is to test the efficacy of triclafascid and two solid dispersion complex preparations based on the triclabendazole substance against fasciolosis in cattle and sheep. Materials and methods . Complex preparations of triclabendazole (TCB) formulations: Succinic Acid (SA): Polyvinylpyrrolidone (PVP) (1: 1 : 8) and TCB: SA: Arabinogalactan (AG) (1: 1 : 8) were obtained by combined solid-stategrinding of the components in a fiber drum on an LE-101 ball mill at a roll rotation speed of 60 rpm with a process module of 1: 17 for 1 to 6 hours with sampling for analysis (HPLC) for changes in the TCB solubility. A comparative test of the efficacy of triclafascid and new solid dispersions (SD) of TCB complexes with SA against fascioliasis of sheep and cattle was conducted in the North-Eastern Federal District of the Caucasus in March-April 2023. To determine the Fasciola infection rate in animals, fecal samples were individually taken and examined by the Fülleborn methods using saturated aqueous NaCL solution and sequential washing. The drug efficacy was recorded at 25-30 days after deworming in analogy with "critical test". The results were statistically processed by the Student-Fisher method using Microsoft Excel 2007. Results and discussion . The resulting TCB and SA complexes with polymers had an increased solubility (up to 59–70 times) that depended on the polymer nature. The efficacy of new TCB dosage forms against fascioliasis in sheep at a dose of 1.5 mg/kg, i.e. 0.5 mg/kg less than the therapeutic dose as compared with oral triclafascid was 90.0 and 100%, respectively, and 85.7% in cattle at a dose of 2.0 mg/kg for the active substance.

The purpose of the research is to study the infection of diurnal birds of prey of the order Falconiformes with helminths and to determine the helminth species composition by the example of the Noyev Kovcheg Conservation Nature’s Nursery in the Altai Territory. Materials and methods . We studied diurnal birds of prey of the order Falconiformes for helminthosis in the Altai Territory, in the Noyev Kovcheg Conservation Nature’s Nursery, in 2022. For the parasitological study of birds, we used laboratory methods of helminthocoprological studies: helminthoovoscopy and helminthoscopy. Species identification was determined by characteristic morphological features of helminth eggs, larvae and fragments. To assess the infeсtion with certain types of helminths, we used standard indicators for parasitological research, namely, infection prevalence and infection intensity amplitude. Results and discussion . In 58 studied diurnal birds of prey of the order Falconiformes, 8 helminth species were identified of which 6 nematode species, namely, Ascaridia galli, Tetrameres sobolevi, Capillaria caudinflata, Heterakis gallinarum, Trichostrongylus spp., and Singamus trachea ; 1 trematode species, Strigea spp.; and 1 cestode species, Raillietina echinobothrida . These species occurred throughout all seasons of 2022.

Among the most common and dangerous parasitic diseases of animals and humans is fascioliasis. The most effective against Fasciola is triclabendazole ("Fasinex 5.0%" suspension), imported by Siba-Geigi, which is practically not used in Russia due to its high cost. Considering the huge economic damage caused by fascioliasis, we have developed a new complex preparation using innovative mechanochemical technology based on the substance of triclabendazole "Triclafascid", which is 5 times more active than the substance of triclabendazole against fascioliasis. The drug is an intermolecular complex in the form of an easily flowing solid dispersed powder with a size of 0.1–10 microns. The drug is administered orally once to cattle and sheep for therapeutic and prophylactic purposes for acute and chronic fascioliasis, to sheep at a therapeutic dose of 2.0 mg/kg and to cattle 2.5 mg/kg for the AI, respectively, and 20 and 25 mg /kg for the drug, once individually in the form of an aqueous solution. In case of mass deworming of animals, it is convenient to give the anthelmintic by a group method to 10–20 animals, mixed with mixed feed at a dose of 3.0 mg/kg for the AI (for the preparation 30 mg/kg).

Deworming untamed herd horses is a difficult task. It is more convenient to use drugs by free feeding, individually or in groups in a mixture with concentrated feed. In this regard, we have developed a new dosage form for the treatment and prevention of parasitosis in herd horses by free feeding in a mixture with concentrated feed based on the substance ivermectin, which has a high antiparasitic activity and a wide spectrum of action. To obtain an antiparasitic supramolecular complex based on the substance ivermectin, an innovative mechanochemical technology was used using two water-soluble polymers of low molecular weight polyvinylpyrrollydon-17 and natural arabinogalactan. The aim of the work is to develop a methodology for the use of the supramolecular complex ivermectin "Aniverm-2.0%" against parasitosis of herd horses. The drug "Aniverm-2.0%" is a supramolecular complex of ivermectin, a water-soluble solid powder of a light beige color, tasteless, with an easily perceptible coniferous odor. The methodology provides a brief description of the drug, its mechanism of action, pharmaco-toxicological properties, the procedure for use, the timing of slaughtering animals and personal safety measures. According to the degree of impact on the body of warm-blooded animals, the drug "Aniverm-2.0%" belongs to low-toxic substances.

The purpose of the research is to detect long-term effects of repeated oral administration of supramolecular complex of ivermectin Aniverm-2.0% on the postnatal development of rat offspring. Materials and methods . The studies were conducted on 16 white pregnant rats that were divided into two experimental and one control groups. The animals were kept under standard conditions of keeping and feeding. The supramolecular complex of ivermectin Aniverm-2.0% was administered to pregnant female rats (n = 6) of group 1 in the form of a suspension using an intragastric tube daily for 7 days at a dose of 15 mg/kg, and group 2 was given the substance of ivermectin (n = 5) at a dose of 8.25 mg/kg. The control female rats (n = 5) were administered 1 mL of distilled water during the experiment. The experimental pregnant females were left until delivery, and then, the development of their offspring was monitored for 45 days. After rat pups were born, the following were recorded: pregnancy duration, litter size, dynamics of weight gain in the rat pups for 21 days, postnatal death during the first 30 days, the ratio of males and females in the litter, periods of eye opening, incisor eruption, detachment of the auricle, appearance of hair coat, descent of testicles, and opening of the vagina. Then we assessed the maturation rate of sensory-motor reflexes in the offspring obtained from the experimental and control groups, the emotional motor behavior and ability for fine coordination of movements in the offspring, and conducted the open field-2 test on day 45 after birth. Results and discussion . No negative effect was detected for supramolecular complex of ivermectin Aniverm-2.0% on the parameters of physiological development of the offspring of the experimental rats within 45 days after birth. The dynamics of their mass, developmental parameters, and formation of motor reflexes during the feeding period remained within the normal values. Developmental indicators of sensory motor reflexes in the control and experimental rat pups had no statistically significant differences.