RU2846896C1 - Antimicrobial preparation and a method for production thereof - Google Patents

Abstract

FIELD: medical science; veterinary science.

SUBSTANCE: group of inventions refers to medicine and veterinary science, namely to broad-spectrum antimicrobial preparations. Disclosed is an antimicrobial preparation containing colloidal silver, benzyldimethyl-[3-(myristoylamino)propyl]-ammonium chloride, isopropanol or ethanol, as well as polyvinylpyrrolidone of molecular weight 25000-3500000 amu with following content of components, wt. %: colloidal silver 0.001-0.05; benzyldimethyl-[3-(myristoylamino)propyl]-ammonium chloride 0.002-0.05; isopropanol or ethanol 1-70; polyvinyl pyrrolidone 0.5-5.0; treated water – balance. Treated water used is water which has passed through a reverse osmosis filter, which is poured into a container, at the bottom of which there is a layer of shungite with particle size of 4-8 mm, and held therein for 24-30 hours. What is also presented is a method for preparing the above preparation.

EFFECT: preparing the effective antimicrobial preparation which can be used in treating endometritis in highly productive cows and characterized by high stability.

2 cl, 10 tbl, 3 ex

Description

FIELD OF TECHNOLOGY TO WHICH THE INVENTION RELATES

The group of inventions relates to medicine or veterinary science, namely to broad-spectrum antimicrobial drugs, as well as methods for their production.

LEVEL OF TECHNOLOGY

The preparation "Argovit" is known from the prior art; it is a dispersion of a complex of colloidal silver with low-molecular polyvinylpyrrolidone. According to TU No. 9310-13-00008064-00 for the preparation, it has a broad spectrum of antimicrobial action against gram-positive and gram-negative, aerobic and anaerobic, spore-forming and asporogenous bacteria in the form of monocultures and microbial associations, including antibiotic-resistant hospital strains. It exhibits high virucidal (antiviral) and fungicidal (antifungal) activity; has a pronounced anti-inflammatory effect; stimulates reparative processes in the wound and on the skin. The silver content in argovit, calculated for 100% dry preparation, is 5-7%. [http://vector-vita.narod.ru/argovit.htm].

The disadvantage of the drug "Argovit" is the relatively high concentration of silver required to achieve a disinfectant effect.

The prior art discloses an antibacterial preparation based on colloidal silver suspended in water, which additionally contains benzyldimethyl-[3-(myristoylamino)propyl]-ammonium chloride [RU2419439, IPC A61K 33/38, A61K 31/14, A61P 17/02, published 27.05.2011]. The aim of the invention is to develop a more effective broad-spectrum antibacterial preparation.

A disadvantage of the above-described antibacterial drug is the insufficiently high stability of the colloidal system, i.e., the possibility of aggregation of silver particles during long-term storage of the drug and the tissues of living organisms, for example, humans or animals, which leads to the precipitation of metallic silver and inactivation of the drug.

An antibacterial composition is known based on colloidal silver stabilized with citric acid or its salts, suspended in water, additionally containing benzyldimethyl-[3-(myristoylamino)propyl]-ammonium chloride [RU2480203, IPC A61K 31/00, A61K 33/38, A61K 31/14, A61K 31/19, A61P 31/04, published 05/27/2011].

The patent document also discloses a method for producing an antibacterial composition based on colloidal silver, in which the following steps are performed: mixing an aqueous solution of a silver salt, a solution of at least one stabilizing agent, and a solution of a reducing agent, while the mixture is vigorously stirred, then a solution of at least one cationic surfactant is added to the resulting mixture.

The aim of the invention is to develop a more effective broad-spectrum antibacterial composition.

The closest to the claimed invention is a disinfectant [RU2427380, IPC A61K 33/38, A61K 31/14, A61L 2/16, A61P 17/02, published 08/27/2011] in the form of a concentrate for treating skin, containing colloidal silver, at least one cationic surfactant, auxiliary additives and water in the following ratio of components, wt. %:

colloidal silver 0.1-0.5 cationic surfactant 1.0-25.0 auxiliary additives 1.0-10.0 water up to 100,

In this case, the concentrate is diluted before use.

The aim of the invention is to develop a more effective disinfectant for antiseptic treatment of skin. This goal is achieved primarily by exploiting the synergistic disinfectant effect that occurs in mixtures of colloidal silver with cationic surfactants (CSAs).

A method for producing a disinfectant according to one embodiment of the invention is also known from a patent document, in which 10 ml of an aqueous solution of silver nitrate (0.2 g, 1.18⋅10-3 mol) are added with stirring to 100 ml of a 0.1% solution of benzyldimethyl[3-(myristoylamino)propyl]ammonium chloride. The resulting mixture is stirred for 15 min, after which 90 ml of an aqueous solution containing 0.1 g ( ^2.6⋅10-3 mol) of sodium borohydride NaBH ₄ and 0.1 g ( ^2.25⋅10-4 mol) of benzyldimethyl[3-(myristoylamino)propyl]ammonium chloride are added with stirring. After adding the entire amount of sodium borohydride, the system is stirred for 1 hour. Then, 1.0 g of sodium citrate (monosodium citrate) is added to the solution, followed by 5 ml (0.5 wt%) of polyethylene glycol (PEG-9) and 45 ml (4.5 wt%) of propylene glycol. This results in a disinfectant concentrate. Before use, the concentrate is diluted 10-fold with distilled or drinking water, resulting in a disinfectant suitable for use on skin.

DISCLOSURE OF THE ESSENCE OF THE INVENTION

The objective of the proposed group of inventions is to expand the arsenal of disinfectants for the antiseptic treatment of animal tissues, including colloidal silver and cationic surfactants, as well as methods for their production.

The technical result is the creation of an effective antimicrobial drug used, in particular, in the treatment of endometritis in high-yielding cows.

An associated technical result also includes obtaining a drug with a stable composition.

The technical results are achieved by the fact that an antimicrobial preparation is proposed that contains colloidal silver, benzyldimethyl-[3-(myristoylamino)propyl]-ammonium chloride, while it additionally contains isopropanol or ethanol, as well as polyvinylpyrrolidone with a molecular weight of 25,000-3,500,000 amu with the following content of components, wt. %:

colloidal silver 0.001-0.05 benzyldimethyl-[3-(myristoylamino)propyl]-ammonium chloride 0.002-0.05 isopropanol or ethanol 1-70 polyvinylpyrrolidone 0.5-5.0%. prepared water rest

A method for producing an antimicrobial preparation is also proposed, in which an aqueous solution containing colloidal silver and benzyldimethyl-[3-(myristoylamino)propyl]-ammonium chloride is mixed with other components of the preparation, characterized in that isopropanol or ethanol is first loaded into the reactor, followed by adding to it, while stirring with a stirrer at a speed of 50-100 rpm, a pre-prepared aqueous solution containing polyvinylpyrrolidone with a molecular weight of 25,000-3,500,000 amu, after which a pre-prepared aqueous solution containing colloidal silver and benzyldimethyl-[3-(myristoylamino)propyl]-ammonium chloride is added to the resulting solution, followed by stirring the resulting solution for 15-20 minutes. with a stirrer speed of 50-100 rpm at a medium temperature of 15-25°C, and for the preparation of aqueous solutions, water is used that has passed through a reverse osmosis filter, poured into a container, at the bottom of which a layer of shungite with a particle size of 4-8 mm is laid out, and kept in it for 24-30 hours.

IMPLEMENTATION OF THE INVENTION

Below are examples of the invention implementation.

Example 1. 70 g of isopropanol are pumped into a 150 ^cm3 reactor. While stirring with a stirrer at 100 rpm, 15 g of an aqueous solution containing 5.0 g of polyvinylpyrrolidone with a molecular weight of 25,000 amu are added to the isopropanol from dispenser #1. 15 g of an aqueous solution containing 0.05 g of colloidal silver and 0.002 g of benzyldimethyl-[3-(myristoylamino)propyl]-ammonium chloride are added to the resulting solution from dispenser #2. The resulting solution is stirred for 15 min at a stirrer speed of 100 rpm at a medium temperature of 15°C.

After mixing is complete, the resulting antimicrobial product is packaged. Water for the aqueous solutions is prepared as follows: water that has passed through a reverse osmosis filter is poured into a cubic container with a layer of 4mm shungite at the bottom and left to soak for 24 hours.

Example 2. 1 g of ethanol is pumped into a 150 ^cm3 reactor. While stirring with a stirrer at 50 rpm, 15 g of an aqueous solution containing 0.5 g of polyvinylpyrrolidone with a molecular weight of 3,500,000 amu is added to the ethanol from dispenser #1. 84 g of an aqueous solution containing 0.001 g of colloidal silver and 0.05 g of benzyldimethyl-[3-(myristoylamino)propyl]-ammonium chloride are added to the resulting solution from dispenser #2. The resulting solution is stirred for 20 min at a stirrer speed of 50 rpm at a medium temperature of 25°C. After stirring is complete, the resulting antimicrobial drug is packaged. Water is prepared for the preparation of aqueous solutions as follows: water that has passed through a reverse osmosis filter is poured into a cubic container, at the bottom of which a layer of shungite with a particle size of 6 mm is laid, and left to stand for 30 hours.

Example 3. 20 g of isopropanol are pumped into a 150 ^cm3 reactor. While stirring with a stirrer at 60 rpm, 15 g of an aqueous solution containing 3.0 g of polyvinylpyrrolidone with a molecular weight of 1,000,000 amu are added to the isopropanol from dispenser #1. 65 g of an aqueous solution containing 0.03 g of colloidal silver and 0.01 g of benzyldimethyl-[3-(myristoylamino)propyl]-ammonium chloride are added to the resulting solution from dispenser #2. The resulting solution is stirred for 17 min with a stirrer speed of 60 rpm at a medium temperature of 20°C.

After mixing is complete, the resulting antimicrobial product is packaged. Water for the aqueous solutions is prepared as follows: water that has passed through a reverse osmosis filter is poured into a cubic container with a layer of 8mm shungite at the bottom and left to soak for 28 hours.

The achievement of the technical result in terms of the antimicrobial action of the drug by implementing the invention according to the above examples is confirmed by the results of the study of antimicrobial activity.

The study was conducted using the methods in accordance with the Guidelines "Methods of laboratory research and testing of disinfectants to assess their effectiveness and safety" (R 4.2.3676-20), taking into account the requirements contained in the "Uniform sanitary-epidemiological and hygienic requirements for goods subject to sanitary-epidemiological supervision (control)" (Section 20. Basic requirements for disinfectants, disinfestation and deratization agents), and "Standard indicators of safety and effectiveness of disinfectants subject to control during mandatory certification" No. 01-12 / 75-97.

The following strains of microorganisms were used in the study: Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus (no. 906), E. coli ATCC 10536.

To eliminate the bacteriostatic effect of the product, a universal neutralizer was used (tween-80-3%, saponin - 3%, histidine - 0.1%, cysteine hydrochloride - 0.1%).

Working cultures were grown on nutrient media: GRM-agar, staphylococcus-agar, produced by the Federal State Budgetary Scientific Institution State Research Center of Applied Microbiology and Biotechnology for 24-48 hours at a temperature of +37°C.

To obtain a bacterial suspension, the bacterial culture was washed from the surface of the nutrient media and diluted in a physiological solution to a concentration according to the turbidity standard, corresponding to two billion microbial bodies in 1 ml.

Antibacterial activity was assessed using test objects. The test objects included linoleum, painted wood, plastic, glass, metal, and tile surfaces contaminated with test microorganisms. The surfaces were disinfected by wiping.

The criterion for the effectiveness of an antimicrobial drug as a skin antiseptic is established as a reduction in surface contamination by at least 99.99%.

The study was conducted in accordance with R 4.2.3676-20. 3.5. Disinfection. Methods of laboratory research and testing of disinfectants to assess their effectiveness and safety. Guidelines (approved by Rospotrebnadzor on December 18, 2020).

The results of the comparison of the effectiveness of the drug "Miramistin" and an antimicrobial drug in the disinfection of test objects contaminated with P. aeruginosa, S. aureus, E. coli are presented in Table 1.

Table 1. Efficiency of the disinfecting action of the drug "Miramistin" and the antimicrobial drug.

Type of microorganism Disinfection time, min Colony-forming unit count (CFU) Disinfection efficiency, % Before processing after processing The drug Miramistin Antimicrobial drug obtained according to examples 1, 2, 3 (average value) P. aeruginosa 0.5 (3.1±0.3) × 10 ⁶ 59±9 74±11 99.99 1 0 0 100 2 0 0 100 S.aureus 0.5 (1.1±0.2) × 10 ⁶ 8±2 6±1 99.99 1 0 0 100 2 0 0 100 E. coli 0.5 (3.8±0.3) × 10 ⁶ 3±1 4±1 99.99 1 0 0 100 2 0 0 100

The results of a comparative evaluation of the effectiveness of the studied preparations in surface disinfection indicate their antimicrobial activity against E. coli, S. aureus, and P. aeruginosa.

The achievement of the technical result in terms of the possibility of using the drug in the treatment of endometritis in high-yielding cows was confirmed by experimental studies conducted in the veterinary laboratory, the analytical research laboratory and the Komsomolskoye PZ department of the Pavlovsky district of the Federal State Budgetary Scientific Institution of the Federal Agency for Agricultural Development.

The object of the research was cows of the Priobskoye type of the black-and-white breed in the 2nd-5th lactation in the first 2 months after calving with a milk yield of 4.5-5.0 thousand kg of milk per year, and the subject of the research was an antimicrobial drug obtained according to examples of the invention.

For the treatment of endometritis, Endometramag-Bio, a gel-based antiseptic from the quaternary ammonium group with the addition of propranolol (a uterotonic agent), and Quinasept-gel, a gel-based antiseptic from the quinoxaline group, were used as comparison drugs. The drugs were administered intrauterinely using a Janet syringe and an artificial insemination catheter.

Treatment was carried out according to the following schemes (Table 2).

Table 2 - Treatment regimens for acute (experiment No. 1) and chronic (experiment No. 2) endometritis

Experiment number Group Name of the drug Dose of administration, ml Route of administration Frequency and periodicity of use 1 1st experimental Antimicrobial drug (example 2) 100 intrauterine 5 times,
daily Uteroton 10 intramuscularly 3 times,
every other day 2nd experimental Antimicrobial drug
(example 3) 100 intrauterine 5 times,
daily Uteroton 10 intramuscularly 3 times,
every other day 3rd experimental Antimicrobial drug (example 1) 100 intrauterine 5 times,
daily Uteroton 10 intramuscularly 3 times,
every other day 4th control Endometramag-
Bio 100 intrauterine 4 times,
every other day 5th control Quinasept gel 100 intrauterine 4 times,
every other day Uteroton 10 intramuscularly 3 times,
every other day 2 1st experimental Antimicrobial drug (example 2) 70 intrauterine 3 times,
every other day Uteroton 10 intramuscularly 3 times,
every other day 2nd experimental Antimicrobial drug
(example 3) 70 intrauterine 3 times,
every other day Uteroton 10 intramuscularly 3 times,
every other day 3rd experimental Antimicrobial drug
(example 1) 70 intrauterine 3 times,
every other day Uteroton 10 intramuscularly 3 times,
every other day 4th control Endometramag-
Bio 70 intrauterine 3 times,
every other day

When assessing the therapeutic effectiveness, the following indicators were taken into account: the timing of the first estrus after calving (independence period) and the end of treatment; the percentage of recovered and inseminated animals, conception rate from the first insemination and within 2-3 months.

Hematological studies were carried out in a veterinary laboratory, determining the following parameters:

1) the content of leukocytes and the percentage of lymphocytes, middle cells and granulocytes, the concentration of erythrocytes, hemoglobin - on the hematological analyzer MicroCC-20 Plus (veterinary);

2) leukocyte formula according to the generally accepted method;

3) Bredeck index (the ratio of the number of lymphocytes and band neutrophils), ISI (the index of the ratio of the sum of neutrophils to lymphocytes) [Kryachko O.V., Romanova O.V. Leukocyte intoxication index as a prognostic criterion in the treatment of chronic pathology in horses / O.V. Krchko, O.V. Romanova // Veterinary practice. - 2003. - No. 3. pp. 26 - 28];

4) lysosomal cation test (LCT) of blood neutrophils with calculation of the average cytochemical coefficient (ACC) [Makarenko N.A. Lysosomal test for assessing the level of resistance of the body of cattle / N.A. Makarenko // Veterinary science. - 1988. - No. 5. - P. 26-28.];

5) protein fractions by nephelometric method.

Hematological parameters were determined using a MicroCC-20Vet hematology analyzer. A differential leukocyte count (leukogram) was performed by microscopic examination of Pappenheim-stained blood smears using a MicroOptixMS 300 light microscope.

Below are the results of studies on the effectiveness of the antimicrobial drug in endometritis, as well as acute postpartum endometritis in cows.

When treating cows with experimental and control drugs, the following results were obtained (Table 3).

Table 3 - Efficacy of antimicrobial drug in acute endometritis in cows

Group Number of heads Inde-pendans-
period, days Time from treatment to the first insemination, days Recovered Was inseminated within 14 days Pregnant within 60 days from calving heads % heads % heads % 1st experimental 14 40±3.4 16±3.6 11 78.6 8 57.1 6 43 2nd experimental 15 38±3.4 13±3.6 12 80.0 10 66.7 8 53.3 3rd experimental 20 35±4.1 11±2.9 16 80.0 14 70 12 60 4th control 9 35±4.1 11±2.9 7 77.8 6 66.7 5 56 5th control 8 47±5.3 18±4.8 6 75 4 50 4 50

After using the antimicrobial drug, a rectal examination revealed that the uterus returned to normal in size and tone in 11 cows (78.6%) of the 1st experimental group, 12 cows (80%) of the second experimental group and 16 cows (80%) of the 3rd experimental group, i.e. recovered by 0.8 and 3.6%, 2.2 and

5% more cows were treated than in the groups treated with Endometramag-Bio and Quinasept-gel, respectively. Furthermore, cows in the third experimental group came into heat sooner after calving and were inseminated within 14 days of treatment completion, which was 12.9%, 3.3%, and 20% higher than in the first, second, and fourth groups, respectively.

Within 60 days after calving, 43% of cows in the 1st experimental group were successfully inseminated, in the 2nd - 10.3%, in the 3rd - 17%, and in the 4th and 5th control groups - 13 and 7% more.

Thus, the antimicrobial according to examples 1, 2, 3 showed better efficiency in the recovery of animals in relation to the control groups, while the fertility rates in the case of using the drug according to example 2 turned out to be slightly worse than in the control groups.

When studying the effectiveness of therapy in the treatment of chronic endometritis with an antimicrobial drug according to the embodiments of the invention and the control drug Endometramag-Bio, the following results were obtained (Table 4).

Table 4 - Efficacy of antimicrobial drug in chronic endometritis in cows

Group Number of heads Recovered Was inseminated within 14 days Pregnant women within 90 days of treatment Fertilized from the first insemination heads % heads % heads % heads % 1st experimental 8 7 87.5 3 37.5 6 75 4 57.1 2nd experimental 12 11 91.7 5 41.6 9 75 7 58.3 3rd experimental 15 14 93.3 7 46.6 12 80 9 60 4th control 8 5 50 2 25 4 50 1 16.7

Following treatment, residual purulent discharge was detected in 1 cow in the 1st and 2nd experimental groups and was absent in the 3rd experimental group, while in the 4th control group, it was detected in 3 cows. Six and nine cows (75%) in the 1st and 2nd control groups, as well as 12 cows (80%) in the 3rd control group, became pregnant within 90 days after calving, while in the 4th control group, 25% and 30% fewer cows were impregnated, respectively.

Within 14 days after treatment, 12.5% and 40.4%, 16.6% and 41.6%, and 21.6% and 43.3% more animals were inseminated and fertilized from the first insemination in the experimental groups than in the control group, respectively. The highest recovery rate (93.3%) was observed in the group treated with the antimicrobial agent as described in Example 3, which is 43.3% higher than with Endometramag-Bio.

Thus, it can be noted that the declared antimicrobial drug has a fairly high efficiency in comparison with similar drugs and can be recommended for use in the treatment of endometritis in cows.

The study also examined changes in the hematological profile and some nonspecific resistance indicators. The following results were obtained from the hematological parameters studied during the treatment of acute postpartum endometritis (Table 5).

Table 5 - Assessment of changes in hematological parameters

Indicator Antimicrobial drug according to examples 1, 2, 3 (average value) Endometromag-Bio Quinasept start ending start ending start ending Leukocytes, 10 ¹² 6.5±0.2 7.1±1.4 9.1±0.5 7.5±0.8 7.0±1.2 7.6±0.9 Lymphocytes, % 56.9±4.0 56.0±6.1 62.5±4.4 57.7±4.5 48.5±13.9 52.1±7.6 Average cells, % 18.3±0.2 17.2±2.4 13.2±1.4 17.7±2.3 15.2±2.1 16.6±1.7 Granulocytes, % 24.9±3.9 26.6±3.6 24.3±3.4 24.7±2.9 36.4±11.9 28.5±6.9 Granulocytes/Lymphocytes 0.44 0.47 0.39 0.43 0.75 0.55 Red blood cells,
10 ⁹ 5.4±0.1 5.9±0.1 5.5±0.3 5.3±0.2 5.9±0.4 5.6±0.3 Hemoglobin, g/l 130.4±1.3 117.5±0.4 119.4±3.1 126.9±3.4 134.4±6.4 127.6±6.5

After the treatment, the changes in the number of leukocytes, erythrocytes and hemoglobin were within the normal range. The content of granulocytes at the beginning of the experiment was below the norm in the experimental groups by 56.6%; in the 4th control - by 57.9; in the 5th control - by 36.8%, having increased by the end of the experiment by 7.0; 1.8 and decreased by 21.7%, respectively. Therefore, the granulocyte/lymphocyte ratio increased in the experimental by - 6.8%, in the 4th control - by 10.3%, and in the 5th control - decreased by 26.7%. At the end of the experiment, in the experimental groups, in comparison with the 4th control group, this ratio increased by 17% and decreased in relation to the 5th control group by 8.5%.

The study of the leukocyte formula showed that the number of eosinophils in the experimental groups increased by 13.4% by the end of the experiment, going beyond the normal range, in the 4th control group it decreased by 2.3 times, in the 5th control group it slightly increased by 7.8% within the normal range. The content of band neutrophils exceeded the upper limit of the norm by 2.26; 3.04 and 2.44 times in all groups, respectively, then increased after the experiment in the experimental groups by 30.5%; in the 4th control group - by 51.4% (p < 0.05) and decreased by 19.7% in the 5th control group. The number of segmented neutrophils decreased by 4.1; 7.6; 0.7%, in the experimental and control groups, respectively. The lymphocyte level varied within the normal range (Table 6).

Table 6 - Leukocyte formula and CBC

Indicator Antimicrobial drug according to examples 1, 2, 3 (average value) Endometromag-Bio Quinasept gel start ending start ending start ending Eosinophils, % 8.6±2.2 9.7±2.4 6.7±2.3 2.9±1.4 6.4±3.5 6.9±1.6 Rod-shaped nuclei, % 11.3±3.7 14.7±2.3 8.1±3.2 18.6±2.5* 15.2±0.8 12.2±3.7 Segmented-nuclear, % 21.9±3.0 19.7±2.7 27.1±2.1 19.5±4.6 26.4±3.1 25.7±1.8 Lymphocytes, % 57.9±3.1 55.5±2.9 57.6±3.6 58.1±3.3 51.1±6.9 54.6±5.2 Monocytes, % 0.4±0.2 0.4±0.2 0.5±0.3 1.0±0.4 0.8±0.3 0.6±0.4 Bredeck index 5.13±0.29 3.77±0.43 7.1±0.42 3.13±0.54* 3.36±0.55 4.46±0.7 ISNL 0.57±0.10 0.62±0.09 0.94±0.09 0.65±0.09 0.81±0.21 0.69±0.17 Central Communications Service,
conventional units 0.74±0.07 0.93±0.11 0.67±0.05 0.80±0.11 0.55±0.09 1.17±0.16 Note: * - (p<0.05) compared to the beginning of the experiment.

In the experimental groups where the antimicrobial drug was used, with a decrease in IB by 26.5%, there was an increase in ISNL by 8.2%. After treatment with Endometromag-Bio, IB decreased by 2.3 times (p < 0.05), ISNL by 1.5 times. In the group where the cows were treated with Quinasept-gel, an increase in IB by 33% and a decrease in ISNL by 14.8% were noted. As a result, in all groups the level of non-specific resistance was 2-2.8 times lower than the norm. By the end of the experiment, the ACC increased in all groups by 1.26; 1.19 and 2.1 times, respectively, while in the 5th control group it was 43.9 and 60.9% higher than in the experimental and 4th control groups and was closest to the norm.

The study of the fractional composition of the blood proteins of cows revealed that serum albumins at the end of the experiment decreased by 1.2 and 4.1% in the experimental and 4th control groups, while in the 5th control group - by 23.6%; β-globulins were below the normal limit and slightly decreased by the end of the experiment in all groups; the content of γ-globulins significantly increased in the 5th control group by 26.9%. Thus, stability of the albumin/globulin ratio was noted in the experimental and 4th control groups compared to the 5th control group, where it decreased almost 2 times due to a sharp decrease in the level of albumins and an increase in the amount of γ-globulins, probably due to an imbalance of humoral factors (Table 7).

Table 7 - Effect of drugs on the protein composition of the blood

Indicator Antimicrobial drug according to examples 1, 2, 3 (average value) Endometromag-Bio Quinasept gel start ending start ending start ending Albumins, % 51.8±3.0 51.2±2.0 47.5±1.5 45.5±3.0 52.3±5.7 39.9±4.3 α-globulins, % 11.7±1.3 12.7±1.5 11.4±0.8 12.8±1.8 7.6±0.9 11.6±1.5 β-globulins, % 6.7±1.0 6.1±0.6 8.5±1.3 7.4±2.3 6.9±0.9 6.5±1.6 γ-globulins, % 29.6±2.9 30.7±1.9 33.9±0.1 34.1±4.6 33.3±6.5 42.3±5.5 Albumins/
globulins 1.1±0.1 1.1±0.1 0.9±0.1 0.9±0.1 1.4±0.5 0.7±0.2

As a result, it can be noted that the declared antimicrobial drug had a more prolonged modulating effect on non-specific resistance, in comparison with the control drugs.

A study of hematological parameters following treatment for chronic endometritis revealed that changes in white blood cell (WBC), red blood cell (RBC), and hemoglobin counts were within normal limits. By the end of the study, the granulocyte/lymphocyte ratio had increased by 7.2% in the experimental groups and by 7.5% in the control group. This ratio remained within normal limits in the experimental groups, while in the control group it was 1.26 times lower than normal, indicating that this ratio was more optimal in the experimental groups (Table 8).

Table 8 - Hematological parameters of cows

Indicator Antimicrobial drug according to examples 1, 2, 3 (average value) Endometromag-Bio Norm start ending start ending Leukocytes, 10 ¹² 5.9±0.5 5.2±0.2 7.7±0.6 6.9±1.0 6.6-9.5 Lymphocytes, % 45.2±3.0 43.3±1.7 55.9±3.0 55.4±5.5 40-65 Average cells, % 19.8±3.4 20.9±1.9 16.9±0.7 16.9±2.0 - Granulocytes, % 35.1±2.7 35.9±2.3 27.3±2.6 27.8±4.4 34-44 Granulocytes / Lymphocytes 0.77 0.83 0.49 0.53 0.67-0.85 Erythrocytes, 10 ⁹ 5.2±0.1 5.1±0.1 5.4±0.3 5.0±0.3 4.5-7.5 Hemoglobin, g/l 117.1±6.2 120.4±4.9 122.9±4.3 125.5±3.8 99-129

A study of the white blood cell count after treatment with the claimed antimicrobial drug revealed a 64.3% decrease in eosinophils within the normal range, while in the Endometromag-Bio group, an increase of 62.8% was observed. Band neutrophil counts increased in the first and second groups by 86.9% and 81.8%, exceeding the upper limit of normal by 4.16 and 4.68 times, respectively. Segmented neutrophil counts decreased by 22.1% and 28.6%, respectively. This indicates a regenerative nuclear shift in neutrophils, likely related to the suppression of inflammatory processes in the body.

After treatment of chronic endometritis with an antimicrobial drug, the Bredecka index decreased by 1.85; in the control group, by 2.5 times, with an increase in the ISNL by 10.8 and 30.5%, respectively, compared with the control drug. The CCR index increased by 19.4% in the experimental group, and after using Endometromag-Bio, it decreased by 9.4%, indicating a positive effect on nonspecific resistance of the antimicrobial drug (Table 9).

Table 9 - Leukocyte formula and CBC

Indicator Antimicrobial drug according to examples 1, 2, 3 (average value) Endometromag-Bio Norm, % start ending start ending Eosinophils, % 10.6±2.7 3.8±1.5 6.6±2.0 10.8±2.4 3-8 Band neutrophils, % 11.1±2.4 20.8±3.8 12.9±3.1 23.4±4.2 2-5 Segmented neutrophils, % 21.7±3.5 16.9±2.8 20.9±4.4 15.0±4.3 20-35 Lymphocytes, % 56.5±3.6 58.0±3.8 59.3±3.7 50.4±2.6 40-65 Monocytes, % 0.2±0.2 0.6±0.2 0.3±0.2 0.4±0.3 2-7 Bredeck index 5.10±0.6 2.8±0.4 4.6±0.5 2.2±0.2 8.75-10.4 ISNL 0.6±0.10 0.7±0.10 0.6±0.10 0.8±0.10 0.44-0.54 SCC, conventional units 0.78±0.1 0.94±0.1 0.89±0.0 0.81±0.2 1.37

When analyzing the protein composition of the cows' blood, it was revealed that the content of albumin and α-globulins at the end of the experiment decreased in the experimental groups by 8.2 and 13.9%, and in the control group by 0.5 and 27.7%, respectively; β-globulins decreased by 0.3% in the experimental groups and increased by 20.6% in the control group; the level of γ-globulins increased by 22.2 and 10.4%, respectively. The albumin/globulin ratio after treatment with the declared antimicrobial drug decreased by 17.7%, below the normal limit, in the group where Endometromag-Bio was used, this indicator remained unchanged (Table 10).

Table 10 - Effect of drugs on the protein composition of the blood

Indicator Antimicrobial drug according to examples 1, 2, 3 (average value) Endometromag-Bio Norm start ending start ending Albumins, % 49.1±3.1 45.1±2.2 46.0±2.9 46.2±2.7 38-50 α-globulins, % 13.7±2.0 11.8±1.6 15.9±1.1 11.5±3.2 12-20 β-globulins, % 6.0±1.3 6.1±1.1 6.8±0.9 8.3±1.6 10-16 γ-globulins, % 31.6±1.9 38.6±3.4 31.2±2.6 34.4±5.4 25-40 Albumins/
globulins 1.0±0.1 0.8±0.1 0.9±0.1 0.9±0.1 0.9-1.4

Thus, the claimed antimicrobial drug obtained by the claimed method has a sufficiently high efficiency in comparison with similar drugs, which allows for the implementation of the invention with the achievement of technical results.

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Claims (4)

1. An antimicrobial preparation containing colloidal silver, benzyldimethyl-[3-(myristoylamino)propyl]-ammonium chloride, characterized in that it additionally contains isopropanol or ethanol, as well as polyvinylpyrrolidone with a molecular weight of 25,000-3,500,000 amu with the following component content, wt.%: colloidal silver 0.001-0.05 benzyldimethyl-[3- (myristoylamino)propyl]-ammonium 0.002-0.05 isopropanol or ethanol 1-70 polyvinylpyrrolidone 0.5-5.0% prepared water rest, Moreover, the prepared water is water that has passed through a reverse osmosis filter, poured into a container with a layer of shungite with a particle size of 4-8 mm at the bottom, and kept in it for 24-30 hours. 2. A method for producing an antimicrobial preparation according to claim 1, which involves mixing an aqueous solution containing colloidal silver and benzyldimethyl-[3-(myristoylamino)propyl]-ammonium chloride with other components of the preparation, characterized in that isopropanol or ethanol is first loaded into the reactor, followed by adding to it, while stirring with a stirrer at a speed of 50-100 rpm, a pre-prepared aqueous solution containing polyvinylpyrrolidone with a molecular weight of 25,000-3,500,000 amu, after which a pre-prepared aqueous solution containing colloidal silver and benzyldimethyl-[3-(myristoylamino)propyl]-ammonium chloride is added to the resulting solution, followed by stirring the resulting solution for 15-20 minutes at a stirrer speed of 50-100 rpm at a medium temperature of 15-25°C, and for the preparation Water solutions are made using water that has passed through a reverse osmosis filter, poured into a container with a layer of shungite with a particle size of 4-8 mm at the bottom, and kept in it for 24-30 hours.