RU2620548C1 - Method for calves respiratory diseases prevention - Google Patents

Abstract

FIELD: veterinary medicine.

SUBSTANCE: clinically healthy calves 20-30 days of age are given xymedon hydrochloride mixed with a 3.5% alcohol tincture of scotch pine (Gemmue Pini) buds, herbs and inflorescences of Ehinacea purpurea L., rhizomes and roots of elecampane (Inula Neleium L.), taken in the ratio 2:1:1, calculated based on 2.0 g of xymedon hydrochloride to 100.0 ml of 3.5% alcohol tincture of scotch pine (Gemmue Pini) buds, herbs and inflorescences of Ehinacea purpurea L., rhizomes and roots of elecampane (Inula Neleium L.), dosed as 1.5-2.0 ml/kg of body weight for the first time 18-24 hours before and twice after immunization. The immunization is carried out three times by subcutaneous injection of hyperimmune serum containing antihaemagglutinin to viruses PG-3 in titer of 1:1280, IRT - in titer of 1:256 and VD-BS - in titer of 1:1024, adenovirus - in titer of 1:256, PC-virus - in titer of 1:128, dosed as 1.0 ml/kg body of weight at an interval of 10-12 days.

EFFECT: higher preventive effect.

4 tbl, 3 ex

Description

The invention relates to veterinary medicine, relates to a method for the prevention of infectious diseases and can be used to prevent massive respiratory diseases of calves.

The etiology of respiratory diseases is complex and includes the adverse effects on the body of calves of various stress factors. At the same time, the indicators of general non-specific resistance and immunobiological reactivity are reduced, as a result of which the body's adaptive capabilities to biological effects (associates of pathogens of a viral, bacterial, chlamydial-mycoplasma nature) and abiotic factors are sharply limited. Prevention of respiratory viral diseases of newborn calves 20-31 days of age has its own characteristics. As a result of the metabolic decay of colostral antibodies and a simultaneous increase in blood volume in calves, a gradual loss of passive immunity occurs. There is a decrease in both cellular and humoral immunity with the underdevelopment of their own immunity. An immunological examination of clinically healthy calves of 19-31-day-old age of an economy unsuccessful for respiratory diseases revealed signs of functional insufficiency of the immune system [1]. They showed a tendency to a decrease in the functional activity of phagocytic cells, indicators of the T-cell immunity, IgG and IgM levels, as well as the immunological reactivity of the body. Thus, the bactericidal activity of blood serum was 29.1 ± 1.4% compared with 31.3 ± 1.2% in clinically healthy calves from a farm that was happy with respiratory diseases, the lysozyme activity of blood serum was 11.3 ± 0.4 % and 13.01 ± 0.5%; the concentration of total protein is 51.3 ± 1.2 g / l and 50.1 ± 1.2 g / l, respectively.

Currently, the use of immunomodulators is considered as one of the promising ways to prevent respiratory infections [2]. The main requirements for the use of immunopharmacological preparations are their immunomodulating properties, clinically proven high antiviral efficacy, safety, lack of addiction, side and carcinogenic effects. Immunomodulators should not cause excessive sensitization and induction of immunopathological reactions, as well as undesirable additive effects when used together with other pharmaceutical agents. The necessary qualities of the acceptability of the drug are also the predictability of the metabolism scheme and ways of excretion from the body, the possibility of combination with other drugs used for infectious and inflammatory diseases. The pyrimidine derivatives, which have found their application in the treatment of infectious, surgical, neurological, oncological and many other diseases, fully meet the listed requirements and represent a group of the most diverse chemicals with a wide spectrum of pharmacological activity. Analogs of pyrimidines have a broad anti-infectious orientation of the pharmacological action. High activity of these drugs is noted both against viruses, microorganisms, fungi, and against parasites. In recent decades, much attention has been paid to the drug Xymedon, developed by the Institute of Organic and Physical Chemistry. Acad. A.E. Arbuzov Kazan Science Center RAS and Kazan State Medical Institute [3].

Xymedon is 1- (β-hydroxyethyl) -4,6-dimethyl-1,2-dihydro-2-hydroxypyrimidine, has regenerative (activation of regeneration processes with linear wounds of the skin and planar skin defects), anti-inflammatory (regulatory effect on the inflammation process; in the first stage of inflammation, the drug has a protective effect at the cellular level due to membrane-stabilizing, antioxidant, adaptogenic, antibacterial effects, in the second phase of inflammation - a pronounced decongestant effect), immunity incriminating (by inducing the expression of CD2 + molecules, FCgR and C3bR on the membrane of immunocompetent cells, enhancing metabolic processes in the cell due to activation of intracellular enzyme systems and DNA synthesis, which leads to stimulation of T and B immunity systems with an increase in the absolute content of CD4 + and CD19 + cells , initiation of proliferation and differentiation of bone marrow precursors of T-lymphocytes along the helper pathway, increased functional activity of T-lymphocytes, stimulation of the phagocytic activity of neutrophils and macrophages with increased eat completion rate of phagocytosis), antibacterial, antioxidant (membranoprotektivnoe effect on the white blood cells) effects. In addition, the drug helps to increase the number of red blood cells by stimulating hematopoiesis and by increasing their resistance to destruction [4]. To date, considerable material has been accumulated about the increased use of Xymedon in the following areas of clinical medicine: for the prevention and treatment of complications in the postoperative period, especially in patients with a high risk of complications, and in emergency abdominal surgery, in the complex treatment of long-healing wounds and trophic ulcers. The appointment of xymedon is indicated for chronic osteomyelitis, in the recovery period after peripheral nerve injuries, in the complex treatment of burn disease and before autodermoplasty. The drug has been successfully used in the treatment of pneumonia, chronic bronchitis, peptic ulcer of the stomach and duodenum, common and destructive forms of tuberculosis, in dentistry [5]. In veterinary medicine, Xymedon hydrochloride (1- (2-hydroxyethyl) -4,6-dimethyl-1,2-dihydropyrimidin-2-one hydrochloride), an intermediate in the process of obtaining xymedon, was used as a means of increasing natural resistance and preventing gastrointestinal intestinal diseases of calves [6], and in combination with mixoferon (an injection preparation based on recombinant α-interferons) for the prevention of associated calf bronchopneumonia: calves 30-45 days old were injected with xymedon hydrochloride at a dose of 25 mg / kg body weight and mixoferon in the amount of 1 dose per kg of live weight, twice with an interval of 7 days [7]. The known methods provide increased resistance of calves to mixed respiratory infections, however, in such animals, clinical signs of the disease do not appear, and pathogens are released. Viruses in high concentration (4.5-5 lg TCD 50 / ml) are secreted from nasal mucus, contents of the conjunctival sac, saliva and urine. In this regard, there is a wide distribution of pathogens and massive reabsorption of animals [8].

The purpose of the invention is to increase the effectiveness of the prevention of mixed respiratory diseases of calves of viral and bacterial etiology by providing complete protection against the development of a clinically expressed disease and stopping the excretion of viruses and bacteria.

This goal is achieved by using an immunotropic drug inside and an additional injection of hyperimmune serum of animal donors containing anti-hemagglutinins to PG-3 viruses in a titer of 1: 1280, ИРТ in a titer of 1: 256 and VD-BS in a titer of 1: 1024, adenovirus - in titer 1: 256, PC virus - in titer 1: 128.

The immunotropic preparation contains 2.0 g of Xymedon hydrochloride and 100.0 ml of 3.5% alcohol tincture of the ordinary pine buds (Gemmue Pini), grass and inflorescences of Echinacea purpurea (Ehinaceapurpurea L.), rhizomes and roots of Elecampane high (Inula Neleium L.) taken in a ratio of 2: 1: 1.

The rationale for the choice of tincture components was data on the pharmacological properties and chemical composition of medicinal plant materials. Each component contains biologically active substances that act simultaneously on all links of pathogenesis, the symptoms of inflammatory diseases of the respiratory organs, and also have a pronounced antimicrobial effect on pathogenic and conditionally pathogenic microflora.

Biologically active substances of pine kidneys (Gemmue Pini) have disinfectant, phytoncide, expectorant, vitamin and soothing properties, effectively prevent the spread of microbial and viral infections in the body.

The biologically active substances of the herb and inflorescences of Echinacea purpurea (Ehinacea purpurea L.) have immunomodulating activity, reduce the activity of hyaluronidase, which is produced by bacteria and viruses, as a result of which hyaluronic acid is preserved, a substance that forms in the tissues between cells and serves as a barrier against infections [9 ], increase the antimicrobial activity of leukocytes, macrophages and blood neutrophils and increase the synthesis of protective antibodies and endogenous interferon.

Biologically active substances of rhizomes and roots of Elecampane high (Inula Neleium I.) have anti-inflammatory, antimicrobial, bactericidal, antifungal, hemostatic, expectorant properties.

To prepare tinctures, the buds of ordinary pine (Gemmue Pini), grass and inflorescences of purple coneflower (Ehinacea purpurea L.), rhizomes and roots of high elecampane (Inula Neleium L.) are crushed (in dry form) and mixed in a 2: 1: 1 ratio. The raw material is placed in a glass vessel and moistened, pouring the extractor (70% alcohol) in a ratio of 1: 5. Discharge of the finished product is obtained after 7 days.

In accordance with the classification of toxicity of substances according to GOST 12.1.007-88, the tool can be rated as relatively safe, because when determining the parameters of acute toxicity in mice weighing 20.0 g, it showed a concentration of 20% and 40% LD ₅₀ more than 17.8 g / kg body weight, and with intrauterine administration - more than 3.0 g / kg body weight.

Hyperimmune serum is prepared according to the method of N.I. Humpback (1981) from the blood of animal donors from the farm for which it is intended, previously tested for cattle leukemia virus in RID with negative results. Animals are immunized with a multivalent antiviral vaccine. Effective serum should contain antibodies to etiological agents of calf respiratory diseases in the following credits: to PG-3 virus - 1: 1280, ИРТ - 1: 256, VD-BS - 1: 1024, adenovirus - 1: 256, PC - to virus - 1 : 128.

The method is as follows.

For clinically healthy 20-30-day-old calves, an immunotropic preparation is administered three times orally at a dose of 1.5-2.0 ml / kg body weight for the first time 18-24 hours before and twice after immunization, which is carried out three times by subcutaneous injection of hyperimmune serum at a dose of 1 , 0 ml / kg body weight with an interval of 10-12 days.

The prophylactic effectiveness of the method was determined in production experiments under conditions of an economy that was long-term inadequate for respiratory diseases due to the impact on the body of virus associations, pathogenic and conditionally pathogenic microflora against the background of violation of the conditions of keeping and feeding of pregnant cows and calves. Serological studies of the blood serum of calves with respiratory infections revealed antibodies to IRT viruses in 80%, PG-3 in 72.5%, VD-BS in 77.5%, PC in 67.5% and to adenovirus - in 60% of cases. At the same time, antibodies to five viruses were detected in 60% of the samples - ИРТ, ПГ-3, ВД-БС, PC, АД, in 22.5% - to four viruses ИРТ, ВД-БС, PC, АД, in 17.5 %) samples - to three viruses, IRT, VD-BS, PC - infection. In bacteriological studies of pathological material from dead calves 13-15 days after the first examination of blood serum of calves, in which antibodies to five and four viruses were detected during the initial study, conditionally pathogenic microflora was represented, represented by various associations with a predominance of staphylococci - 64.3%, streptococci - 42.9%.

The essence of the method is illustrated by examples.

Example 1. The effectiveness of prophylaxis, depending on the dose of the immunotropic drug, was determined on 5 groups of clinically healthy calves 20-30 days of age: 4 groups - experienced, 1 group - control. The conditions of feeding, care and maintenance are the same in all groups. The calves of all experimental groups were used three times inside the immunotropic preparation in doses of ml / kg body weight of 1.0 (first group), 1.5 (second group), 2.0 (third group) and 2.5 (fourth group) for the first time 18 hours before and twice after immunization, which was carried out three times by subcutaneous injection of hyperimmune serum at a dose of 1.0 ml / kg body weight with an interval of 10 days. The calves of the control group did not use drugs. Clinical status was determined by generally accepted methods; all the dead animals underwent a complete pathological and anatomical autopsy with the sending of pathological material to the veterinary laboratory. The results are presented in table 1.

As can be seen from table 1, doses in the claimed range of 1.5-2.0 ml / kg body weight are necessary and sufficient to obtain optimal indicators of the clinical status of calves.

Example 2. The effect of an immunotropic preparation on the immune status of clinically healthy calves was determined in 3 groups of clinically healthy calves of 20-30 days of age compared with conditional control (clinically healthy calves of 20-30 days of age in a calf respiratory disease safe). The calves of the first group were injected three times with an immunotropic preparation at a dose of 1.5 ml / kg of live weight for the first time 18 hours before and twice after immunization, which was carried out three times by subcutaneous injection of hyperimmune serum at a dose of 1.0 ml / kg of live weight with an interval of 12 days. Calves of the second group were injected with xymedon three times in a dose of 2.0 ml / kg live weight for the first time 24 hours before and twice after immunization, which was carried out three times subcutaneously with hyperimmune serum at a dose of 1.0 ml / kg live weight with an interval of 12 days . Calves of the third group were immunized with a triple subcutaneous injection of hyperimmune serum at a dose of 1.0 ml / kg body weight with an interval of 12 days. The functional state of the immune system was evaluated by indicators of cellular and humoral immunity, including the relative and absolute content of blood T and B lymphocytes (determined by the method of N.I. Blinov, 1980), the functional activity of blood neutrophils in the HCT test (determined by method M .E. Wicksman and AM Mayansky, 1979), immunoglobulin levels of individual isotypes (G and M) (determined by the method of Manchini, 1964), lysozyme activity of blood serum (determined by the method of V.G. Dorofeychuk, 1969). Blood collection from animals for studies was carried out before the use of drugs (background) and 10 days after the completion of the experiment. The results are presented in table 2.

As can be seen from table 2, in all groups noted changes in indicators of immune status relative to baseline values. Higher rates of humoral and cellular factors of natural resistance were provided by the immunotropic preparation in accordance with the invention. So, in comparison with the conditional control, the relative content of T-lymphocytes was as close as possible to the corresponding indicator, and B-lymphocytes exceeded the corresponding indicator, the absolute content of T-lymphocytes exceeded, and the absolute content of B-lymphocytes reached the corresponding indicator. Similar dynamics were noted for other indicators studied.

Example 3. The comparative effectiveness of the proposed method was determined on 4 groups of clinically healthy calves 20-30 days of age: 3 groups - experienced, 1 group - control. The calves of the first experimental group were used three times inside the immunotropic preparation in accordance with the invention at a dose of 2.0 ml / kg body weight for the first time 24 hours before and twice after immunization, which was carried out three times subcutaneously with a hyperimmune serum injection at a dose of 1.0 ml / kg live mass with an interval of 12 hours (the inventive method). The calves of the second experimental group were injected with xymedon hydrochloride at a dose of 25 mg / kg live weight and mixoferon in an amount of 1 dose per kg live weight, twice with an interval of 7 days (prototype method). The calves of the third experimental group were subcutaneously injected with hyperimmune serum of animal donors at a dose of 1 ml / kg of live weight three times with an interval of 12 days (basic method). Calves of the fourth group of drugs were not used (control). All calves were subjected to general clinical examination. 10 days after the end of the experiments, blood was taken to determine the main immunological parameters. The results are presented in tables 3 and 4.

As can be seen from table 3, the prophylactic administration of drugs in all groups was carried out against the background of disorders of the immune system in the form of a decrease in the functional activity of phagocytic cells, a decrease in T-cell link, IgG and IgM deficiency. In all experimental groups, there was a tendency to normalize immunobiological parameters. More pronounced changes of a normalizing nature were established in the first group, especially with respect to IgG and IgM levels. So, the IgG level in calves of the first group increased by 42.06% compared with 32.03% (second group), 17.18% (third group) and 6.67% (control). IgG proteins represent the class of antibodies G. They make up about 80% of all immunoglobulins. IgG antibodies provide long-lasting humoral immunity in infectious diseases, i.e. are antibodies of a secondary immune response to foreign substances. The IgM level in calves of the first group increased by 41.18% compared with 17.69% (second group), 6.25% (third group) and 5.89% (control). IgM - proteins representing the class of antibodies M, the first to produce antibodies in response to acute infection, providing primary immunity. They are the first to appear in the bloodstream with bacteremia, realizing antibacterial immunity. The indicators of functional activity of blood neutrophils in spontaneous and induced tests in calves of the first group were higher by 31, 81% and 64, 08%, respectively, compared to 17.39% and 50.26% in the second group, 13.04% and 37.57% in the third group, 5.88% and 9.04% in the control. An increase in the resistance of the organism of animals of the experimental groups was also reflected in the number of cases (3.1% - the first group, 9.1% - the second group and 7.9% - the third group compared with 13.3% in the control) (table 4). Symptoms of respiratory pathology in calves that were using immunotropic drugs were manifested mainly in a mild form and were characterized by serous rhinitis, a rare dry cough, and an increase in body temperature by 0.5-1.2 ° С. Symptoms of rhinitis persisted for 6-8 days. In the control group, the percentage of patients was high (20.0%), and the disease was difficult, with complications, accompanied by damage to the gastrointestinal tract and relapses. Clinical signs in sick animals were characterized by a short-term increase in body temperature to 40.5-41 ° C, a decrease or lack of appetite, general depression, the appearance of serous-mucous discharge from the nasal cavity, rapid breathing, coughing, conjunctivitis. Affected calves were treated with traditional methods using symptomatic agents. Clinical recovery occurred after 16 days.

Studies have confirmed the high efficiency of the proposed method of prevention using an immunotropic agent based on Xymedon hydrochloride and hyperimmune serum. The components of an immunotropic agent have high bioavailability and a multivalent positive effect on the animal's body. Hydrochloride 1- (2-hydroxyethyl) -4,6-dimethyl-1,2-dihydropyrimidin-2-one (xymedon hydrochloride) is an immunomodulator with a pronounced regenerative and antimicrobial effect. The tincture of the buds of Scots pine (Gemmue Pini), herbs and inflorescences of Echinacea purpurea (Ehinacea purpurea L.), rhizomes and roots of Elecampane high (Inula Neleium L) contains biologically active substances that increase the functional activity of antioxidant and immune systems, in particular, macrophage cells. Passive immunization against the background of correction of immune dysfunctions provides reliable protection of calves not only against infectious rhinotracheitis, parainfluenza-3, viral diarrhea - a disease of mucous membranes, adenovirus and respiratory syncytial infections, but also from a possible association of other pathogenic viruses circulating in livestock farms, including herpes viruses , entero-viruses and numerous unclassified viruses, and the bacteriostatic activity of an immunotropic drug is from pathogens of bacterial origin.

Information sources

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Claims (1)

A method for the prevention of respiratory diseases of calves, including the use of clinically healthy calves 20-30 days of age, xymedon hydrochloride, characterized in that xymedon hydrochloride is used in a mixture with 3.5% alcohol tincture of ordinary pine buds (Gemmue Pini), grass and purple coneflowers (Echinacea purpurea) ( Ehinacea purpurea L.), rhizomes and roots of Elecampane high (Inula Neleium L.), taken in a 2: 1: 1 ratio, based on 2.0 g of xymedon hydrochloride per 100.0 ml of 3.5% alcohol tincture of ordinary pine buds ( Gemmue Pini), herbs and inflorescences of Echinacea purpurea (Ehinacea purpurea L.), rhizomes and roots of elecampane high (Inula Neleium L.) at a dose of 1.5-2.0 ml / kg body weight for the first time 18-24 hours before and twice after immunization, which is carried out three times by subcutaneous injection of hyperimmune serum containing antihemagglutinins to viruses PG-3 in titer 1: 1280, ИРТ - in titer 1: 256, and VD-BS - in titer 1: 1024, adenovirus - in titer 1: 256, PC virus - in titer 1: 128, at a dose of 1.0 ml / kg of live weight with an interval of 10-12 days.