RU2623063C2 - Bovine cattle leukosis prevention and treatment method - Google Patents

Abstract

FIELD: veterinary medicine.

SUBSTANCE: method involves the use of a complex preparation with antiviral and immunostimulating activity. Argotiazine is used as the preparation. The cows are given the drug for therapeutic purposes intramuscularly at a dose of 3 mg/kg of animal weight, 4-fold, with the interval of 4 days. The drug are added to the 6-8 months calves with prophylactical purpose intrarectally at a dose of 20 mg/head in 300 ml of boiled water, once a day for 10 consecutive days.

EFFECT: method is highly effective for the prevention and treatment of the bovine cattle from leukosis.

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Description

The invention relates to the field of veterinary medicine, in particular for the prevention of the clinical and hematological form of lymphoid leukemia in spontaneously infected bovine leukemia virus (VLSC) and the treatment of sick animals, and can also be used in microbiology, virology and immunology.

Bovine leukemia is a widespread chronic viral disease that causes significant economic damage to dairy farming. Treatment and prevention have not been developed to date, therefore, after identifying animals infected with leukemia virus, they are isolated and kept in a separate group. Those infected with leukemia of cows with an increased number of leukocytes / lymphocytes per unit blood volume, identified according to the “leukemia key”, are discarded from the herd in a short time, and the milk received from them is prohibited for use, since altered ones accumulate in the body, as well as in milk. under the influence of the virus, lymphoid cells and metabolic products with carcinogenic properties (Rules for the prevention and control of cattle leukemia, approved by order of the Ministry of Agriculture and Food USSIAN Federation, May 11, 1999 № 359).

Compounds that are capable of exerting an antiviral and immunomodulating effect are known, one of which is a “water-soluble agent having antiviral activity based on a silver compound with cystine and a method for its preparation (RF Patent No. 2277908 of 06/20/2006, IPC A61K 31/197). The disadvantage is the fact that the drug was not developed for the treatment and prevention of viral diseases of animals.

Another analogue of the claimed method is known, "Water-soluble agent with antiviral and immunomodulating activity, based on the combination of ionic silver with methylene blue and the method for its preparation", corresponding to the composition C16H18C12N3SAg with the working name Argotiazine (RF Patent No. 2390343 dated 05/27/2010, IPC A61K 33 / 38, A61K 31/5415, A61K 31/28, A61P 37/02, A61P 31/12), which can be taken as closest to the solution of the proposed method. The compound has antiviral and immunomodulating activity, photostability in aqueous solutions and retains its properties for 2 years of storage. It was previously shown that the combined use of Argotiazine and a new chemical compound πQ346 from the class of physiologically compatible antioxidants with immunostimulating activity, effectively prevents the appearance of hematological signs of leukemia in spontaneously infected VLCRS cows (VL Tikhonov, SG Nikiforova. Methodical recommendations “Development of new antiviral agents based on silver ions and their influence on the pathogenetic process in cattle leukemia.” - Irkutsk, 2010. - 34 p.).

The disadvantage of this tool is the lack of a method for its individual use, including for animals with leukemia of animals with hematological signs of the disease, and its other doses, which allows treating patients with VLCRS infection of animals, leading to the remission of clinical signs of leukemia and preserving highly productive animals from inevitable culling .

The task of the invention is to expand the arsenal of methods for the prevention and treatment of animals with leukemia in cattle, increasing the effectiveness of recreational anti-leukemia measures.

The essence of the invention lies in the fact that in the method for the prevention and treatment of leukemia in cattle, including the use of a complex preparation with antiviral and immunostimulating activity, according to the invention, Argotiazine is used as a preparation, and the drug is administered intramuscularly to cows for therapeutic purposes at a dose of 3 mg / kg animal weights, 4-fold, with an interval of 4 days, and for calves 6-8 months of age, the drug is administered prophylactically rectally at a dose of 20 mg / goal in 300 ml of boiled water, once a day for 10 days in a row.

When developing a method for the prevention and treatment of cattle leukemia, the values of the antiviral and immunomodulating activity of Argotiazin obtained in in vitro and in vivo studies (V.N.Silnikov, V.V. Tretyakov, V.L. Tikhonov and others. Development of Argotiazine and the study of its antiviral activity // J. Siberian Bulletin of Agricultural Science. - 2009. - No. 9. - P. 60-63; T.I. Glotova, V.L. Tikhonov, A.S. Donchenko and others. Study of immunostimulating properties Argotiazina // J. Siberian Bulletin of Agricultural Science. - 2009. - No. 11. - S. 73-77).

In addition, when studying the pharmacokinetics of Argotiazin in animals spontaneously infected with VLCC, after a single intramuscular injection of a drug at a dose of 1 mg / kg of animal weight, we found that the maximum value of mobile silver in the blood and feces of animals fell at 6 hours. In the process of elimination and metabolism of the compound, the silver concentration gradually decreased, reaching the minimum detectable value of silver in the body after 72-96 hours.

In order to preliminary establish the optimal therapeutic and prophylactic doses of Argotiazine and a rational scheme for its use in leukemia of cattle, the following clinical studies were conducted.

Prevention of leukemia in VLSC infected calves 6-8 months of age (latent stage of leukemia) by rectal administration of Argotiazine:

1) 10 mg / goal, once a day for 5 days in a row,

2) 10 mg / goal, once a day for 10 days in a row,

3) 20 mg / goal, once a day for 5 days in a row,

4) 20 mg / goal, once a day for 10 days in a row,

5) 30 mg / goal, once a day for 5 days in a row,

6) 30 mg / goal, once a day for 10 days in a row.

Treatment of cows with viral leukemia by intramuscular injection of Argotiazine:

1) 1 mg / kg, twice with an interval of 4 days,

2) 1 mg / kg, four times with an interval of 4 days,

3) 3 mg / kg, twice with an interval of 4 days,

4) 3 mg / kg, four times with an interval of 4 days,

5) 5 mg / kg, twice with an interval of 4 days,

6) 5 mg / kg, four times with an interval of 4 days.

Monitoring and evaluation of the state of the leukemia process in animals treated with Argotiazine was carried out according to standard diagnostic tests for leukemia in comparison with similar indicators obtained in background studies. The inhibitory effect of Argotiazine on the infectious process in leukemia was evaluated by lowering the titer level of antibodies specific for VLCC during serological testing of blood serum and its dilutions (1: 2-1: 32) in the agar gel immunodiffusion reaction (RID) between 1 and 6 months after termination of the course treatment of animals with a drug compound (Guidelines for the diagnosis of leukemia in cattle. M., 2000. - 34 p.). The RID (+) result of native serum was taken as the minimum antibody titer. In addition, hematological studies of blood were carried out with the calculation of the relative number of leukocytes and the absolute number of lymphocytes in 1 μl of blood, guided by the "leukemia key" (Guidelines for the diagnosis of leukemia in cattle. M., 2000. - 34 p.).

The anti-leukemia efficacy of Argotiazine was evaluated within 6 months of follow-up after treatment of the animals. Studies have shown that the average dose of the drug for calves with a latent form of leukemia, at which a stable preventive effect was achieved after a single daily, for 10 consecutive days, rectal use of Argotiazine, was 20 mg / goal. The five-day use of this dose and its lower value with a 10-fold use did not lead to 100% prophylaxis of the incidence of leukemia in VLCC-infected animals.

In cows with a hematologic form of leukemia, the most pronounced therapeutic effect, characterized by prolonged remission of the clinical signs of the disease (within 6 months), was manifested after four times with an interval of 4 days intramuscular injection of Argotiazine in a dose of at least 3 mg / kg.

Production assessment of the proposed method for the prevention and treatment of leukemia in cattle obtained in the framework of the invention, is illustrated by examples and tables.

Example 1. To prevent the progressive development of leukemia among VLCRS-infected calves of 6-8 months of age born from cows infected with VLCRS with confirmed positive serological diagnosis of leukemia (without clinical and hematological signs), a rectal administration of Argotiazine at a dose of 20 mg / goal was carried out in 300 ml of boiled water using a catheter, 10-fold, with an interval of 24 hours (table. 1).

Before the start of the experiment on the prevention of leukemia in calves of 6-8 months of age, all experimental and control animals were seropositive for the leukemia virus, and in some individuals the titers of antibodies specific for VLCRC reached a level of 1: 8 (No. 89). It’s known that the higher the level of precipitating antibodies in the blood serum of animals infected with leukemia virus, the more intense the infection process is (TR Yakupov. New approaches in the diagnosis of cattle leukemia // Scientific notes of Kazan State Academy of Veterinary Medicine named after N.E.Bauman. - 2010. - No. 1. - vol. 204. - S. 342-347).

At the beginning of the experiment, some calves observed signs of incipient hematological stress (Table 1), from which it can be concluded that animals with developing VLCRS infection were selected for the experiment.

As follows from the table. 1, daily, for 10 consecutive days, rectal use of Argotiazine in calves of the experimental group infected with leukemia virus, at a dose of 20 mg / goal. leads to the containment of the process of further development of the disease. The result of stopping the infection, in comparison with the control group, was observed for 6 months or more after treatment of the animals and was characterized by a 2–4-fold decrease in titers of specific VLCRC antibodies and stabilization of the number of leukocytes / lymphocytes per unit volume of blood within physiological limits.

Since the persistence of colostral antibodies in newborn calves born from mothers that are seropositive for leukemia lasts up to 6 months, a systematic serological assessment of young animals starting at 6 months of age allows for timely assessment of the epizootic situation and the development of a preventive treatment plan for animals that are compromised in relation to the leukemia virus the method of Example 1, thereby reducing the risk of leukemia spreading.

Example 2. The treatment of animals with clinical and hematological signs of viral lymphocytic leukemia was performed by four times, with an interval of 4 days, intramuscular injection of Argotiazin at a dose of 3 mg / kg live weight in 10 ml of sterile distilled water (in 10 ml of sterile 0.9% chloride solution sodium).

A therapeutic dose of the drug was injected into the gluteal muscle group in turn, on the right and left side. The clinical results of the effect of Argotiazine on the infectious process with leukemia are shown in table. 2 and 3.

From table 2 it follows that the intramuscular administration of Argotiazine at a dose of 3 mg / kg to cows with leukemia in the experimental group 1 month after the course of treatment led to a 2–4-fold decrease in titers of VLCC-specific antibodies in comparison with background values. In some animals, a “loss” of the serological reaction was observed and precipitating antibodies were not detected (No. 3892/7 and 841/3). Further serological studies of blood serum and its dilutions (3 and 6 months) confirmed the low content of precipitating antibodies among experimental animals. At the same time, among the control patients with leukemia of cows that did not undergo anti-leukemia therapy, a 2-fold increase in titers of antibodies specific for VLCRC was observed (No. 703/2), which is typical for further progressive development of the leukemia process.

When considering the results of hematological studies on leukemia among cows with leukemia in the experimental group and treated with Argotiazine for therapeutic purposes (Table 3), the following was noted. The use of the drug in patients with leukemia in cows leads to remission of the clinical and hematological signs of leukemia in the 4-week period after completion of the course treatment. Under the influence of Argotiazin, hematologically sick cows with leukemia experienced not only a decrease in the level of precipitating antibodies in the blood serum, but also a significant decrease in the number of leukocytes / lymphocytes to the physiological “normal” limits. The duration of a positive result from a therapeutic procedure was at least 6 months. In the group of control cows with leukemia, all animals dropped out during the experiment period in connection with the development of a pronounced form of clinical leukemia in them.

Thus, bearing in mind that in the Russian Federation there is a constant increase in cattle leukemia-deficient points and the number of animals infected and leukemic infected continues to grow, in the absence of highly active anti-leukemia vaccines and drugs in the arsenal of the veterinary service, the use of the proposed method as preventive and therapeutic therapy, allows you to effectively control the incidence of animals VLKRS infection and directionally control the epizootic process om, thereby reducing the risk of further spread of leukemia.

Claims (1)

A method for the prevention and treatment of cattle leukemia, including the use of a complex drug with antiviral and immunostimulating activity, characterized in that Argotiazine is used as the drug, and the drug is administered intramuscularly to cows for therapeutic purposes at a dose of 3 mg / kg of animal weight, 4 times, with an interval of 4 days, and for calves 6-8 months of age, the drug is administered prophylactically rectally at a dose of 20 mg / goal. in 300 ml of boiled water, once a day for 10 consecutive days.