RU2540428C1 - Method of treatment of brucellosis of cattle - Google Patents

Abstract

FIELD: veterinary medicine.

SUBSTANCE: method comprises administering a mixture of medicinal solution of formaldehyde and isotonic solution of sodium chloride at a ratio by weight of (2-6):(994-998). The mixture is administered intramuscularly at a dose of 5-6 ml 6 times. The second injection is administered on the day 7-8 after the first injection, the third - on the day 7-8 after the second injection, the fourth - 12 hours after the third, the fifth - 12 hours after the fourth, the sixth - 7-8 days after the fifth injection.

EFFECT: method is highly effective in treatment of brucellosis of cattle.

3 tbl, 3 ex

Description

The invention relates to veterinary medicine, in particular to methods for treating infectious diseases of animals, namely brucellosis.

There is a method of treating human brucellosis, which consists in the intramuscular injection of 5 injections of Reaferon-EU at a dose of 1 million ME every 72 hours and then oral administration of the drug Cycloferon 0.6 g once a day for 1, 2, 4, 6, 8, 11 , 17, 20, 23rd days (see RF patent No. 2423143, class IPC A61K 38/21, publ. 07/10/2011).

Also known are methods of treating human brucellosis with antibiotics: chloramphenicol, tetracycline, a combination of tetracycline with streptomycin (see, for example, http://www.amny.ru/13_brucel_lech.html).

Known methods for treating human brucellosis and using vaccines, for example, a vaccine based on a purified protective antigen, which is a protein-polysaccharide complex isolated from Brucella abortus or Brucella melitensis species strains in S-form (see RF patent No. 2076493 for cl. IPC A61K 39/10, publ. 03/27/1997).

However, all known methods are applicable only in medicine and are used to treat a person.

Closest to the claimed is a method for the treatment of brucellosis (RF patent No. 2234311, class IPC A61K 9/127, publ. 08/20/2004), including oral administration of liposomes with an antibiotic, while pefloxacin or azithromycin is introduced as an antibiotic, and additionally an immunomodulator - lycopid or levamisole, which is included in pairs with the indicated antibiotic in the total liposomal vesicle.

However, this method, as well as the foregoing, is not applicable for the treatment of cattle.

It is well known and as noted by a number of researchers (see, for example, http://www.allvet.ru/diseases/all_inf16.php; http://xreferat.ru/55/1301-1-profilaktika-i-psihicheskie-rasstroiystva- pri-brucelleze-brucellez-u-hivotnyh.html) that brucellosis in animals is considered an almost incurable disease. This is due to powerful allergic (HRT) and autoimmune reactions, which even after elimination (removal) of the pathogen have a pathological effect on the body. In addition, brucella can pass into the L-form, in the form of which they parasitize for a long time (sometimes for years) in the hosts.

Thus, the treatment of brucellosis in animals, in particular cattle, is not currently developed. Numerous therapeutic agents tested for these purposes have not yielded positive results.

The invention is aimed at solving the problem of creating a method for the treatment of brucellosis in cattle.

To solve the problem in a method of treating brucellosis, which consists in administering a medicament, according to the invention, a medicament containing a mixture of a medical solution of formaldehyde and isotonic sodium chloride solution at a weight ratio of (2-6) is used as a medicine: (994-998), in this case, the drug is administered intramuscularly at a dose of 5-6 ml 6 times, and the second injection is administered on the 7-8th day after the first injection, the third on the 7-8th day after the second injection, the fourth - 12 hours after the third, the fifth - cn contractible 12 hours after the fourth, sixth - 7-8 days after the fifth injection.

The sources of patent and scientific and technical information known to the authors do not describe a method for treating cattle brucellosis based on a drug containing small concentrations of formaldehyde, which is administered according to a certain scheme selected experimentally.

The authors experimentally established that the claimed drug administration scheme provides immunological rearrangement in relation to the pathogen in infected cows.

In this case, the first and second injections reduce the activity of the lymphocytic immunity, which leads to a decrease in the allergization of the body and does not interfere with the completed phagocytosis of brucella.

The third injection, carried out 7 days after the second, and then the subsequent two injections (fourth and fifth), carried out at intervals of 12 hours from each other, ensure the digestion of the pathogen in infected phagocytic cells.

The sixth injection of the drug, carried out 7 days after the 5th injection, allows you to enhance the effect obtained, ensuring the elimination of the pathogen from the animal’s body and fixing the therapeutic effect.

Thus, the authors revealed a new property of a mixture containing a medical solution of formaldehyde and an isotonic sodium chloride solution at certain ratios of parts by weight, to regulate the immunological rearrangement of the animal organism in a certain frequency of administration.

In the inventive method, the task of completely freeing the body of the cows from the intracellular parasite, which until now has been impossible, has been solved.

The foregoing allows us to conclude that there is a criterion of "inventive step" in the claimed invention.

The formaldehyde medical solution is a clear, colorless liquid with a peculiar pungent odor, miscible with water and alcohol in any ratio. The concentration of the active substance (see, for example, http://www.tiensmed.ru/news/formalin-f6s.html) is 36.5--37.5%.

Formaldehyde is a representative of the HCNO aldehyde class. It is a colorless gas with a pungent odor, they say. weighing 30.03, its density at 20 ° C is 0.815, the melting point is 92 ° C, and the boiling point is 19.2 ° C. It is soluble in water, alcohol. It polymerizes easily, forming paraformaldehyde during polymerization in an aqueous medium, and polyoxymethylene in anhydrous environments (butane, hexane).

Isotonic sodium chloride solution for injection is a colorless, clear, salty taste liquid of 0.85-0.95% concentration. The solution is sterile, pyrogen-free.

Sodium chloride - cubic crystals or white crystalline powder, salty taste, odorless. Soluble in water (1: 3).

The finished product in the form of a mixture of a medical solution of formaldehyde with an isotonic solution of sodium chloride is a clear, colorless liquid, odorless, slightly salty taste.

The drug is prepared as follows.

Take 2-6 parts by weight of a medical solution of formaldehyde, add it to 998-994 parts by weight of a sterile solution of sodium chloride for injection. The product is stored in a dark place at a temperature of 15-35 ° C.

The method is as follows.

Take the prepared preparation in the form of a mixture of a medical solution of formaldehyde with an isotonic sodium chloride solution at a ratio of parts by weight (2-6) :( 994-998).

The drug is administered as follows:

1 injection - 1st day;

2 injection - on the 7-8th day after the first injection;

3 injection - on the 7-8th day after the second injection (either on the 14th or on the 15th day after the first);

4 injection - 12 hours after the 3rd injection;

5 injection - 12 hours after the 4th injection;

6 injection - 7-8 days after the fifth injection.

Example 1

An example implementation of the method

The method was tested in one of the dysfunctional brucellosis farms in 70 adult sick cows.

The diagnosis was made based on the results of generally accepted serological tests: animals responded positively to the agglutination reaction (RA) and the long-term complement fixation reaction (RDSK).

A preparation containing 2 weight parts of a medical solution of formaldehyde and 998 weight parts of a sterile 0.85-0.95% sodium chloride solution for injection was used as a therapeutic drug.

The processing of adult animals was performed six times according to the following scheme:

- first injection - in 1 day,

- second injection - on the 7th day after the first injection,

- the third injection - on the 7th day after the second injection (or on the 14th day after the first),

- the fourth injection - 12 hours after the third,

- fifth injection - 12 hours after the fourth,

- sixth injection - 7 days after the fifth injection.

On the 7th day after the last injection, blood was taken from all adult animals and examined serologically for brucellosis, while serological blood tests were performed in RA and RDSK. The results of serological studies of cows are shown in table 1.

Table 1 The results of serological blood tests of cows Total Researched Reacted positively Reacted negatively before drug administration after a 6-fold administration of the drug RA RDSK RA RDSK RA RDSK goals % goals % 70 70 70 27 36 43 61.4% 34 48.6%

As can be seen from table 1, out of 70 goals that previously responded positively to animal brucellosis in RA and RDSK before the drug was administered, in the second study after drug administration, 27 goals in RA and 36 goals in RDSK reacted positively.

Healthy animals were registered in the agglutination reaction of 43 heads, which is 61.4%, and in the reaction of long-term complement fixation, 34 heads, which is 48.6%.

Observations of seronegative cows showed that they subsequently did not have positive reactions in RA and RDSK, which indicated the recovery of the animals (observation period - 3 months).

In the following examples, data are presented to justify the mechanism for implementing the proposed method at the cellular level - on splenocytes and peritoneal cells of white rats.

Example 2

Evidence of a decrease in the activity of the lymphocytic immunity in white rats

Previously, the concentration of the preparation for cell culture was developed, which was the ratio of the weight parts of the medical solution of formaldehyde to the isotonic sodium chloride solution as 3: 166, i.e. in dilution of the main drug 600 times.

Six white outbred rats were used in the experiment, splenocytes were isolated from each of them, suspended in DMEM nutrient medium to a concentration of 1.4 million cells / μl. 200 μl of saline was added to the 1800 μl suspension and incubated at 37 ° C for 40 minutes.

Then, two samples of 800 μl each were taken from the suspension. 100 μl of saline was added to one of them and it served as a further control, and to another, 100 μl of the drug diluted 600 times. After that, 100 μl of 0.1% nitro-blue tetrazolium (HCT) was added. The resulting splenocyte cultures were incubated at 37 ° C for 2 hours, the accumulation of formazan in ng / 10 ⁶ cells was determined. Table 2 presents the results of the HCT test in rat splenocytes.

table 2 The results of the determination of the HCT test in rat splenocytes Animal number The number of formazan in rat splenocytes, ng / million cells control (saline) the drug in the ratio of weight parts 3: 166 one 2,51 1.83 2 2,30 2.05 3 2,38 2.03 four 2.05 1.78 5 2.78 1.94 6 2.49 2,39 Average 2.40 ± 0.1 2.0 ± 0.1

From table 2 it follows that in all samples of splenocytes taken from 6 animals, a significant (p <0.01) decrease in the accumulation of formazan in rat splenocytes was recorded, i.e. lymphocyte activity in the presence of the drug decreased by 16.7% compared with the control.

Example 3

Evidence of activation of phagocytic cells in white rats

A preparation was used, analogously to example 2, in a dilution of 600 times, i.e. with a ratio of components 3: 166.

In the experiment 3 white outbred rats were used, from which peritoneal cells were isolated according to the standard method. Cells were suspended in DMEM with 10% fetal bovine serum to a final cell concentration of 2 million cells / ml.

Then, two samples of 800 μl each were taken from the suspension. 100 μl of saline was added to one of them and it served as a further control, and to another, 100 μl of the drug diluted 600 times. After that, 100 μl of 0.1% nitro-blue tetrazolium (HCT) was added. The resulting splenocyte cultures were incubated at 37 ° C for 2 hours, the accumulation of formazan in ng / 10 ⁶ cells was determined. Table 3 presents the results of the HCT test in rat peritoneal cells.

Table 3 The results of determining the HCT test in rat peritoneal cells Animal number The number of formazan in rat peritoneal cells, ng / million cells control (saline) the drug in the ratio of weight parts 3: 166 one. 15,218 16.58 2. 11.89 12.83 3. 11.15 12.77 Average 12.75 ± 2.16 14.06 ± 2.18

From table 3 it follows that in all 3 cases, an unreliable increase in the activity of peritoneal (phagocytic) cells was recorded (on average by 10.3%). This allows us to conclude that the drug does not affect the activity of phagocytic cells, but causes their release from the pathogen, predetermining the therapeutic effect.

The obtained results prove that the claimed method, based on the treatment of cows according to a certain scheme with a preparation containing a mixture of a medical solution of formaldehyde with an isotonic sodium chloride solution, provides immunological rearrangement of the organism of infected animals, contributing to the elimination of the pathogen and providing a therapeutic effect.

Claims (1)

A method of treating cattle brucellosis, which consists in administering a drug, characterized in that the drug containing a mixture of a medical solution of formaldehyde and isotonic sodium chloride solution in a ratio of parts by weight (2-6) :( 994-998), is used as a medicine in this case, the drug is administered intramuscularly at a dose of 5-6 ml 6 times, with the second injection being administered 7-8 days after the first injection, the third - 7-8 days after the second injection, the fourth - 12 hours after the third, and the fifth - after 12 hours After the fourth, the sixth - in 7-8 days after the fifth injection.