RU2430722C1 - Agent for prevention and treatment of mastitis in cows - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention concerns veterinary science. An agent for prevention and treatment of mastitis in cows contains cetyltrimethylammonium chloride (CAC) or cetyltrimethylammonium bromide (CAB), cuccinic acid, polyhexamethylene guanidine (PHMG) chloride or PHMG phosphate and water in the following proportions, wt %: PHMG chloride or PHMG phosphate 0.5-1.0; CAC 0.05-0.5; succinic acid 0.05-0.5; water - the rest.

EFFECT: invention provides wider spectrum of biocide action, improved antimicrobial and preventive activity of the agent combined with lower skin irritant properties, and also prolonged action.

4 tbl, 1 ex

Description

The invention relates to the field of agriculture, in particular to veterinary medicine, and can be used for the treatment and prevention of mastitis in cattle.

Mastitis in cows is widespread and causes great economic damage to livestock. Animals can get sick at any time of the year, at different times of lactation and during the dead time.

The problem of combating microorganisms that cause infectious diseases in humans and animals is very relevant, despite a fairly large selection of biocidal drugs. Currently, several thousand chemical compounds with biocidal properties are known, but in practice, for safety reasons, only hundreds of them are used. Every year, dozens of biocidal drugs are discontinued due to their low antimicrobial activity or high toxicity. In addition to the natural resistance of some microorganisms to biocidal drugs, microorganisms quickly adapt to adverse factors, including the effects of antimicrobial agents. Given this, as well as the increasing demand for environmental safety of drugs, their toxicity and allergenicity, there is a constant need to search for fundamentally new environmentally friendly biocidal drugs.

The most relevant directions in the creation of new biocidal agents are not so much an increase in their antimicrobial activity (since their toxicity tends to increase as a rule), but an increase in the duration of their antimicrobial activity after surface treatment, a decrease in toxicity, allergenicity and environmental safety. Of great importance is also a wide range of biocidal effects of drugs in relation to pathogenic microflora, acceptable physicochemical, hygienic and consumer properties.

Known drug for the treatment of prophylaxis of bovine mastitis, comprising an antibiotic - ristomycin, lanomin and petrolatum (Patent RF №2135166, Cl A61K ^31/00, 1999, at BI №24..).

However, the use of a drug containing an antibiotic has several disadvantages: dysbiosis, prolonged excretion of antimicrobial substances with milk, and therefore milk is unsuitable for food purposes.

The closest to the proposed invention is the antiseptic agent include quaternary ammonium salt (QAS), mineral or organic acid and a solvent (US Pat. RF №2118174, cl. A61K ^2/16, publ. 1998 YG).

As a QAS, cetyltrimethylammonium bromide or cetyltrimethylammonium chloride, or N-alkyl-N-alkoxycarbonylmethyl-hexahydroazepinium chloride, or cetylpyridinium bromide, or cetylpyridinium chloride, or 1,2-ethylene dimethyl dimethylbimethyl or dialkyldimethyl ammonium chloride, or alkyldimethylbenzylammonium chloride at a certain ratio of components.

However, the known tool is not effective enough in relation to pathogens of intestinal infections and other representatives of gram-negative microflora - constant and transient contaminants of various environmental objects, including integumentary tissues of animals.

The aim of the invention is to expand the spectrum of biocidal action, increase the antimicrobial and prophylactic activity of the drug while reducing irritating properties in relation to integumentary tissues, as well as an increase in prolonged action.

This goal is achieved by the fact that in an agent for the prevention and treatment of mastitis, including cetyltrimethylammonium chloride (QAS) or cetyltrimethylammonium bromide (QAC), organic acid and water, succinic acid is used as the organic acid and polyhexamitylene guanidine chloride (PHMGG) or phosphorus are added in the following ratio of components, wt.%:

PHMG chloride or PHMG phosphate 0.5-1.0 HOUR 0.05-0.5 succinic acid 0.05-0.5 water rest

The invention is illustrated as follows.

PHMG salts - PHMG chloride and PHMG phosphate are well-known substances for the production of disinfectants and skin antiseptics and are very easy to handle: they are non-aggressive, form a monomolecular protective layer, have no odor, do not irritate the skin, are water soluble, have little toxicity (IV class) possess a prolonged effect (> 10 days) and a wide spectrum of antimicrobial action (virucidal, bactericidal, fungicidal), stable during storage. They are approved for use in the food industry, medical practice and in everyday life.

PHMG chloride and PHMG phosphate are produced by ROO IETP under the trademark BIOPAG and FOSFOPAG.

Entering into a synergistic interaction with other active substances, in this case with HOUR, PHMG enhances the biocidal effect of the drug.

Quaternary ammonium bactericidal compounds are used as antiseptic agents for treating surgeons' hands, surgical field, and bladder sanitation (V.I. Vashkov. Means and methods of sterilization used in medicine. M., "Medicine", 1973, p. 216-220 ), for the prevention and treatment of chronic runny nose through prolonged systematic irrigation of the nasopharynx, as well as for the treatment of infected wounds (G.E. Afinogenov, P.I. Blinov. Antiseptics in surgery. L., "Medicine", 1987, p.75- 90 and others; T. Franklin, J. Snow. Biochemistry of antimicrobial action. M., " ir ", 1984, p.72).

The combined interaction of HOUR and PHMG has a stronger bactericidal effect than each of these disinfectants separately. In addition, the spectrum of antimicrobial activity is expanding significantly, and due to the formation on the udder of a molecular polymer film of polyguanidine, the antimicrobial preservation period increases several times.

Succinic acid is a natural substance - present in all organisms. Succinic acid stimulates the production of energy in cells, neutralizes free radicals, has a powerful antimicrobial property, and strengthens the immune system.

As a part of the product, succinic acid regulates the ionic strength of the solution.

Succinic acid (C ₄ H ₆ O ₄ - GOST 6341-75) is a product obtained from the processing of natural amber. It is a white, crystalline powder that resembles citric acid to taste.

The tool is obtained by mixing the components with water.

The quantitative content of the components of the tool is optimal and is selected on the basis of numerous experiments.

If the composition of the PGMG compound contains more than 1.0 wt.%, The cost of the agent increases, and if it is less than 0.5 wt.%, The prolonged effect decreases and the disinfectant effect of the agent decreases.

The content of HOUR in the composition of the product more than 0.5 wt.% Will lead to a deterioration in consumer properties of the product, and its content of less than 0.05 wt.% Leads to a deterioration of the technological properties of the tool.

An increase in the succinic acid content of more than 0.5 wt.% Increases the cost of the product, and a decrease of less than 0.05 wt.% - reduces the therapeutic effect.

The invention is illustrated by the following examples.

Example No. 1

By mixing the components, the following composition is prepared, wt.%:

PHMG chloride 0.5 cetyltrimethylammonium chloride 0.2 succinic acid 0.1 water rest

The shelf life of the prepared product is 2 years.

Example No. 2

Similarly prepare the following composition, wt.%:

PHMG chloride 1,0 cetyltrimethylammonium chloride 0.05 succinic acid 0.5 water rest

The prepared composition is suitable for 2 years.

Example No. 3

Similarly prepare the following composition, wt.%:

PHMG phosphate 0.7 cetyltrimethylammonium bromide 0.05 succinic acid 0.05 water rest

The prepared composition is suitable for 2 years.

Example No. 4

Similarly prepare the following composition, wt.%:

PHMG phosphate 1,0 cetyltrimethylammonium bromide 0.2 succinic acid 0.3 water rest

The prepared composition is suitable for 2 years.

To determine the antimicrobial activity, samples were used according to examples No. 1, 2, 3, 4, and the prototype composition was adopted as a control sample:

cetyltrimethylammonium bromide 2.0 wt.% boric acid 2.0 wt.% water rest

The studies were carried out according to the Guidelines "Methods for testing the antimicrobial activity of preventive antiseptics No. 11-13-1-97" on the basis of LLC Rusmedeko Company.

To determine the antimicrobial activity of the samples, sanitary-indicative microorganisms were used: Staphylococcus aureus, Escherichia coli, Proteus mirabilis, Mycrococcus luteus, Candida albicanas, Bacillus subtilis. Microbial suspensions were prepared according to a turbidity standard of 5 × 10 CFU / cm ³ . In a sterile tube, 1 cm ^{3 of the} test sample pre-heated to 37 ° C was taken and a bacterial suspension of microorganisms (each separately) was added thereto in an amount of 1 cm ³ and kept in a water bath at the same temperature for 30 minutes, then this mixture was neutralized with 3% tween -80 for 10 minutes. The neutralized mixture of culture with the agent in an amount of 1 cm ^{3 was} transferred to Petri dishes and poured into MPA (12-15 cm ³ ), cooled to 40 ° C, and kept in a thermostat for 24 hours (37 ° C). At the same time, control plates were placed with cultures diluted with an equivalent amount of sterile distilled water.

To establish the effect of protein load on the level of antimicrobial activity, 20% horse serum was used.

The results of studies in a quantitative suspension method with respect to the above culture strains showed that the studied samples of the agent in their native form with a microbial load of 5 × 10 ⁵ CFU / cm ³ and an exposure of 30 minutes ensured a decrease in microbes by more than 5 lg. In this case, the protein load did not significantly affect the effectiveness of the drug. In the control, a similar situation was observed.

The irritant effect of the proposed tool was studied on 10 rabbits according to the "Methodological guidelines for determining the toxic properties of drugs used in veterinary medicine and animal husbandry" - Reference "Veterinary drugs", ed. Tretyakova A.D. - M .: Agropromizdat, 1988. S. 242-243. The means according to Example No. 4 were applied to the trimmed areas of the skin of experimental rabbits, and the control rabbits received the remedy according to the prototype in doses of 2-3 g per head for 21 days. During the experiment, we observed the reaction of the skin of rabbits (hyperemia, soreness, thickening of the skin fold, scratching), as well as the general clinical condition of the animals. During the experiment, the skin of the rabbits was painless, without thickening, scratching, hyperemia, as in the control, i.e. the product did not have irritating properties.

The allergenic properties of the agent were determined according to the “Guidelines for the determination of the toxic properties of drugs used in veterinary medicine and animal husbandry”, the reference book “Veterinary drugs”, ed. Tretyakova A.D. - M .: Agropromizdat, 1988, S.244-246. Guinea pigs weighing 200 g, 10 animals were used. The animals were sensitized by repeated (14 days) applications of the product (according to example No. 4) on the same clipped skin area. Then, 14 days after the last application, a resolving dose of the agent (native agent) was applied to the skin of the freshly cut area. After applying the product, the animals remained within the physiological norm for 24 hours; the skin was unchanged.

To conduct tests on the effectiveness of the proposed product on cows, two groups of lactating cows were formed (experimental and control). The nipples of the udder of the cows of the experimental group were treated with the proposed tool, and the control group with the composition according to the control sample (prototype).

The products were applied immediately after milking on the nipples of the udder of the cows. Before milking, the product was thoroughly washed off.

To study the bacteriostatic effect of the agents, swabs were taken from the surface of the nipples of the udder from an area of 10 cm ² before treatment before the next milking, 30 minutes, 2 and 3 hours after treatment with the agent. The studies were carried out by the method of successive dilutions with further seeding on nutrient media, incubation in a thermostat and taking into account the number of grown colonies.

The physicochemical properties of milk (density, acidity, total protein, fat, total bacterial contamination, somatic cells and inhibitory properties) were determined according to TU 00028493.380-98 “Cow's milk. Purchase Requirements. ”

When studying the bacteriostatic effect of the proposed tool, it was found that the total bacterial contamination of the skin surface of the nipples of the udder of the cows of the experimental and control groups decreased 2 hours after treatment.

Table 1 Antimicrobial Test Results for Lactating Cows Group of cows Before processing.
Quantity, CFU / cm ² In 2 hours.
Quantity, CFU / cm ² Experienced Example No. 1 15,000 350 Example No. 2 13800 150 Example No. 3 14800 270 Example No. 4 13600 180 Control 14250 3600

table 2 The results of diagnostic studies of cows for the incidence of mastitis after the use of funds Indicators Group of animals experienced (as in example No. 4) control to experience end of experience to experience end of experience Surveyed, goal. 73 73 74 74 Revealed: Clinical mastitis, goal. one 0 one 0 Clinical mastitis,% 1.4 0 1.4 0 Subclinical mastitis, goal. 9 one 8 6 Subclinical mastitis,% 12.3 1.4 10.8 8.1 Irritation of the udder, goal. 5 0 6 four Irritation of the udder,% 6.8 0 8.1 5,4

It was found that after applying the proposed tool, the skin of the nipples of the udder was soft, elastic. Allergic reactions, irritation was not observed. Existing scratches, erosion, skin lesions in the experimental groups healed within 5-7, the control - 9-10 days.

The results of studies of the physicochemical properties of milk are presented in table 3. From the above data it can be seen that the use of funds did not adversely affect the quality of milk. Received milk from experimental animals corresponded to the highest grade.

Table 3 Physico-chemical characteristics of milk after application Indicators Group experienced (as in example No. 4) control Density, kg / m ³ 1031.0 1030.5 Acidity, ⁰ T 18.0 17.8 Total protein,% 3.11 3.00 Somatic cells, thousand / cm ³ 408.0 413.5 Inhibitory properties neg. neg. General bacterial contamination (class) one one The relative biological value,% 100,4 100.0

The results show that the proposed tool has a good level of antimicrobial activity and meets the requirements of SanPiN 21-112-99 "Disinfectants and technologies. Regulatory indicators of the safety and effectiveness of disinfectants. "

The agent has a bacteriostatic effect in relation to cultures of Staphylococcus aureus, Escherichia coli, Proteus mirabilis, Mycrococcus luteus, Candida albicanas, Bacillus subtilis.

The proposed tool has bacteriostatic, wound healing, emollient properties, does not have an allergenic and irritating effect on the body of cows, does not adversely affect the quality of milk, and helps reduce breast diseases with subclinical mastitis.

Claims (1)

An agent for the prevention and treatment of mastitis in cows, including a quaternary ammonium compound (QAS) - cetyltrimethylammonium chloride or cetyltrimethylammonium bromide, organic acid and water, characterized in that succinic acid is used as an organic acid and additionally contains polyhexamethylene guanidine chloride or PHMGD or in the following ratio of components, wt.%:
PHMG chloride or PHMG phosphate 0.5-1.0 HOUR 0.05-0.5 succinic acid 0.05-0.5 water rest