RU2464999C1 - Method for immunoprophylaxis of respiratory diseases in calves - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to veterinary science. A method for immunoprophylaxis of respiratory diseases in calves involves immunisation with an associated vaccine for parainfluenza-3, infectious rhinotracheitis and bovine viral diarrhoea viruses in a combination with the use of an alcoholic solution of a herbal raw material as an agent for non-specific prophylaxis with the herbal raw material presented by a mixture of equal proportions of purple Echinacea herb and blossom clusters, Syrian rue herb, tillet blossom and common licorice roots which is imbedded in 70% ethanol in relation 1:10, kept in a dark place at temperature 18-20°C for 7 days; the prepared tincture is prescribed orally in the form of 7-8% aqueous solution 1.5-2.0 ml/kg of animal's body weight for 15 days with immunising calves of 60 days old.

EFFECT: invention provides higher effectiveness of immunoprophylaxis.

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Description

The invention relates to veterinary medicine, relates to the prevention of infectious diseases of young farm animals and can be used for the prevention of respiratory diseases of calves proceeding as mixed infections.

Respiratory diseases of calves, which coincide with age-related immunodeficiency, affect up to 50% of nascent animals and cause a loss of up to 40% of the number of cases, taking into account death and forced slaughter. The causative agents of such infections are the viruses belonging to the families of herpes viruses (infectious rhinotracheitis virus), paramyxoviruses (parainfluenza-3 viruses and respiratory syncytial virus), togaviruses (diarrhea virus - mucosal diseases), adenoviruses, rota- and coronaviruses, etc. These are the so-called "small" infections, which in healthy animals with normal functioning of the immune system are asymptomatic without pronounced clinical signs, or animals do not get these infections at all. Animals are especially seriously ill when two or more viruses are involved in the pathological process, that is, a mixed infection occurs. The course of respiratory diseases develops in two phases: the first is the viral phase, the second is the bacterial one. In severe cases of the viral phase of the infection, along with the defeat of sensitive cells, significant inhibition of cellular and humoral immunity occurs. A characteristic feature of the spread of infectious rhinotracheitis viruses, viral diarrhea, and respiratory syncytial infection is that these pathogens can cause enteritis in young animals up to 1 month old, respiratory diseases in older animals, and reproductive dysfunction in adult mature cows and bulls. In addition, parainfluenza-3 virus is widespread in calves older than one month of age with breaches of growing technology, i.e. under stressful effects on the body, causing damage to the respiratory tract, and rota- and coronaviruses in newborn calves cause severe leaky enteritis. According to tropism, all of the above pathogens can be reproduced in the cells of the respiratory and gastrointestinal tracts, genitals, immunocompetent cells, which makes them highly contagious and the severity of the disease.

One of the triggers for infecting animals with infectious diseases in modern industrial technology is a decrease in the immunological reactivity of the body, which is caused by a number of factors - the underdevelopment of the immune system of young animals (primary immunodeficiency), foodborne toxicosis, insufficient and unbalanced feeding of various components, and severe stress the so-called "technological stress." These factors negatively affect the immune system and metabolic processes of the animal organism, which leads to a significant decrease in resistance to infectious diseases, especially to diseases caused by opportunistic microorganisms and viruses. A depressed immune system under the influence of the above factors is not able to withstand pathogens, even with low pathogenicity. The viruses that cause respiratory infections in cattle, each individually, but especially in associations, in turn, inhibit individual parts of the immune system, both cellular and humoral. Along with suppression of the immune system, sensitive cells of the respiratory tract and gastrointestinal tract are affected, which are a good substrate for the propagation of opportunistic microflora. Thus, viruses, breaking the protective barrier of the mucous membranes, contribute to their colonization by bacterial pathogens, so any infection in the animal’s body most often develops not in a mono-but in an associated variant with others.

Numerous antibiotics are used to treat and prevent respiratory diseases. More than 30 thousand items have already been created and their new modifications are constantly being created. However, in the vast majority of cases, antibiotic therapy is prescribed on an empirical basis, which does not always bring the proper therapeutic effect. This becomes understandable, given that antibiotics do not have a destructive effect on viruses. Microorganisms quickly adapt and subsequently unhindered cause bacterial complications. In addition, with prolonged use of antibiotics, they accumulate in the animal organism, which contributes to the emergence of a secondary immunodeficiency state.

The search for protection against the associated complex of viral and bacterial pathogens that cause respiratory pathology is carried out in three directions: 1) passive immunization with specific hyperimmune sera; 2) active immunization with live attenuated or inactivated vaccines; 3) increase the nonspecific resistance of the body by the introduction of immunotropic drugs.

Immunization of calves with specific hyperimmune sera requires significant costs and time / 1 /. In addition, they can be contaminated by numerous pathogens, in particular cattle leukemia virus. Passive immunization can be overcome by highly virulent infectious rhinotracheitis viruses in association with adenovirus and respiratory syncytial infections.

Known methods for the immunological prophylaxis of viral respiratory diseases of calves using associated vaccines, for example live attenuated vaccine "TRIVAC" (VIEV) / 2 / or emulsion inactivated vaccine (ARRIAH) / 3 /. The use of live or inactivated vaccines does not prevent the latency of virulent viruses in animals.

The use of numerous immunotropic drugs is also a serious problem for livestock. So, long-term use of immunostimulants, immunomodulators or immunocorrectors causes the activation of T- and B-cell immunity. This can subsequently lead to the launch of tumor transformation of cells, the mechanism of which is the same as the activation of cells of the immune system (Ageenko A.I., 1992).

The greatest protective effect is achieved with the combined use of vaccination and non-specific immunoprophylaxis / 3 /. The complex system of non-specific immunoprophylaxis includes phytoadaptogens with an immunostimulating effect, for example, derivatives of eleutherococcus, licorice root / 4 /, echinacea purpurea / 5 /.

The technical result is an increase in the effectiveness of immunization.

The technical result is achieved by a combination of vaccination and non-specific immunoprophylaxis using tinctures of grass and purple coneflower Echinacea mixed with common harmonica grass, small-leaved linden flowers and licorice roots taken in equal amounts.

The ingredients used are widely used in herbal medicine. The proposed collection contains all the necessary substances, the combined use of which has an effective effect on the animal's body with respiratory diseases.

The claimed collection has a bronchodilator effect, promotes sputum discharge, has anti-inflammatory and soothing effects, exhibits antiviral and antimicrobial activity, helps restore the functions of the bronchopulmonary system, has detoxification properties, regulates metabolic processes and increases the body's resistance.

The grass and inflorescences of Echinacea purpurea have immunomodulatory activity, inhibit the activity of hyaluronidase, an enzyme that enhances the permeability of viruses and microbes, increase the antimicrobial activity of leukocytes, macrophages and blood neutrophils and increase the synthesis of protective antibodies and endogenous interferon.

The grass of harmonica vulgaris has a pronounced antimicrobial activity, confirmed by pharmacological studies, provides a bronchodilator action of the collection, promotes the discharge of sputum.

Small-leaved linden flowers have immunomodulating effects, which are manifested in the action on the humoral and cellular mechanisms of immunity, lymphocyte migration, interferon synthesis and complement system, and anti-inflammatory effect.

The roots and rhizomes of licorice naked provide bronchodilator and anti-inflammatory action of the collection. In addition, they enhance the healing effects of other herbs included in the collection.

The chemical composition of the therapeutic and prophylactic agent is represented by a complex of biologically active substances providing its high pharmacotherapeutic effect. The dominant biologically active substances are: phenolic compounds, flavonoids, polysaccharides, alkaloids, tannins, vitamins, trace elements, the main of which can be considered zinc, selenium and iron.

Flavonoids (containing all components) inhibit the enzyme catechol-ortho-methyltransferase and provide the adaptive-trophic function of the body due to the adrenoprolonging effect. Flavonoids have P-vitamin (capillary-strengthening) activity. In the total anti-inflammatory effect of flavonoids, their direct effect on cell and tissue membranes is important - a decrease in their permeability, densification of lysosome membranes, inhibition of hyaluronidase, antagonistic effect against inflammatory mediators.

Polysaccharides (contain all components) have an anti-inflammatory, immunocorrective effect. They stimulate nonspecific immunity and phagocytosis, the activity of macrophages and neutrophils, as well as bactericidal function of cells.

Phenolic compounds (containing all components) have an antimicrobial effect, the mechanism of which is due to their ability to be sorbed by the components of the cytoplasmic membrane of bacteria, form strong hydrogen bonds with proteins and damage the membrane, in particular increasing its permeability to ions (primarily potassium) and important metabolites lost cell, as well as for water coming from the outside and contributing to lysis.

78% of the total amount of alkaloids contained in the grass of common harmonica is peganin. Pharmacological studies have shown that 1% and 10% infusions of common harmonica grass have high antistaphylococcal activity, and a 0.25% peganin solution has high antimicrobial activity.

Tannins (naked licorice - up to 9.46%, small-leaved linden - not less than 8.0%), exhibiting a membrane-stabilizing effect, restore repair processes in hepatocytes and improve the functioning of the antioxidant system of the liver.

Vitamins (vitamin C - small-leaved linden flowers, purple coneflower; vitamin E - naked licorice, vitamin A - purple coneflower) are involved in all metabolic processes, increase the body's resistance to diseases, have anti-inflammatory effects, and contribute to detoxification of the body.

Trace element zinc (small-leaved linden - 29 μg / g, naked licorice - 0.33 μg / g, harmala vulgaris - 0.78 μg / g) is involved in the body's immune processes, selenium (naked licorice - 12.14 μg / g, common garmala - 1.0 μg / g, small-leaved linden - 0.13 μg / g) is a powerful antioxidant, and iron (purple coneflower - 48 mg / 100 g, small-leaved linden - 0.2 mg / g, ordinary garmala - 0 , 14 mg / g, licorice - 0.7 mg / g) - the main structural component of hemoglobin of blood and heme-containing enzymes.

High collection efficiency is achieved due to the combination of a whole complex of biologically active substances, as a result of which there is a mutual strengthening of the beneficial pharmacological properties of each individual component. So, the combination of vitamins A, C and zinc stimulates the synthesis of antibodies, the combination of flavonoids and tannins provides a hepatoprotective effect, and the combination of flavonoids and phenologic glycosides enhances the detoxification function of the liver. The collection has a pronounced anti-inflammatory, antimicrobial, antiviral, bronchodilator and adaptogenic effects.

The tincture from the crushed plant mixture of grass and purple coneflowers, common harmonica grass, small-leaved linden flowers and licorice roots is prepared in the pharmacopeia method in a ratio of raw material to extractor 1:10. Ethanol with a concentration of 70% is used as an extractor. Medicinal raw materials are crushed to particles with a size of 1-3 mm, mixed in equal amounts, a weighed portion of the raw material is placed in a glass vessel, the calculated amount of ethyl alcohol at a concentration of 70% is added, it is kept for 7 days in a dark place at a temperature of 18-20 ° C. Then the alcohol tincture is filtered through a canvas or two layers of gauze, squeezing the raw materials, filtered.

Example: 500 g of medicinal raw material were placed in a glass jar (equal amounts from the herb and inflorescences of Echinacea purpurea, common harmonica grass, linden blossoms and licorice roots), 5.0 kg of ethyl alcohol at a concentration of 70% were added, put in a dark place and kept at a temperature of 18-20 ° C for 7 days, after which it was filtered through a canvas, squeezing the raw materials, and filtered.

The method is as follows.

Clinically healthy calves of 60 days of age are immunized with a double injection of the associated vaccine against PG-3, IRT, VD-BS (vaccine dose - 2.0 ml, the interval between injections - 21 days) and an additional 70% alcohol tincture from the crushed plant mixture equal amounts of grass and inflorescences of purple coneflower, common harmonica grass, small-leaved linden flowers and licorice roots in the form of 7-8% aqueous solution prepared in boiled chilled water, at a dose of 1.5-2.0 ml / kg of animal body weight in within 15 days.

The effectiveness of the method for the prevention of respiratory diseases of calves of a polyetiological nature was determined in scientific and production experiments under conditions of a long-term adverse respiratory disease caused by the exposure of the organism to virus associations, pathogenic and conditionally pathogenic microflora against the background of violation of the conditions of keeping and feeding of pregnant cows and calves. Virological and bacteriological studies have confirmed the etiological role of PG-3, IRT, VD-BS, pasteurell, Salmonella, Proteus and Escherichia viruses. Incidence - 60-80%, mortality - up to 20%.

The essence of the method is illustrated by examples.

Example 1. Prophylactic efficacy depending on the dose was determined on 4 groups of clinically healthy calves of 60 days of age. Calves of all groups were immunized intramuscularly in the middle third of the neck with two injections with an interval of 21 days between injections of the emulsion inactivated vaccine against parainfluenza-3, infectious rhinotracheitis and viral diarrhea, a mucosal disease at a dose of 2.0 ml. For 15 days, the calves of the first three groups (experimental) were additionally administered orally 20-30 minutes before feeding with alcoholic tincture from a crushed plant mixture of equal amounts of grass and purple coneflowers, common harmonica grass, small-leaved linden flowers and licorice roots as 7% aqueous solution prepared in boiled chilled water, at doses of 1.5, 2.0 and 2.5 ml / kg of animal body weight, respectively. Animals of the fourth group did not receive additional drugs and served as a control. The effectiveness of prevention was evaluated by the main clinical indicators (table 1).

Table 1 The effectiveness of the prophylaxis of respiratory diseases of calves, depending on the dose of non-specific prophylaxis Indicators Groups of animals experienced control immunization by intramuscular injection in the middle third of the neck with double injections with an interval of 21 days between injections of the emulsion inactivated vaccine against parainfluenza-3, infectious rhinotracheitis and viral diarrhea - mucosal diseases at a dose of 2.0 ml. 70% alcohol tincture from a crushed plant mixture of equal amounts of grass and purple Echinacea inflorescences, common harmonica grass, small-leaved linden flowers and licorice roots in the form of a 7% aqueous solution, in a dose, ml / kg body weight no drugs used 1,5 2.0 2.5 Number of animals, goal. 25 26 24 eighteen Ill, goal. (%) 2 (8) 2 (7.7) 2 (8.3) 2 (16.7) It’s fallen, a goal. 2 Efficiency% 92.0 92.3 91.7 83.3

An increase in the effectiveness of the prevention of respiratory diseases of calves in the first and second groups compared with the control group by 8.7 and 9.0%, respectively, confirmed the validity of the claimed dose of non-specific immunoprophylaxis.

Before vaccination, 21 days after vaccination and 21 days after revaccination, blood was examined in calves of the second experimental and control groups to assess the immune status and level of natural resistance. The immune status was assessed by the relative and absolute number of T- and B-lymphocytes (determined by the method of N.I. Blinov, 1985), the content of the main classes of immunoglobulins IgG, IgM (determined by the method of Mancini et al., 1965) and the level of virus-neutralizing antibodies to PG-3 viruses, IRT, VD-BS in the indirect hemagglutination reaction; natural resistance - by indicators of the functional activity of blood neutrophils (determined in the reaction of tetrazolium recovery according to A.N. Mayansky et al., 1979), bactericidal activity of blood serum (determined by the method of O.V.Smirnova and T.A. Kuzmina, 1966) and lysozyme activity of blood serum (determined by the method of O.V. Bukharin and others, 1970). The results are presented in table 2.

As can be seen from table 2, the means of non-specific prophylaxis in accordance with the invention increased post-vaccination immunity as a result of activation of cellular and humoral immunity. So, on the 21st day after vaccination, the relative and absolute number of blood T-lymphocytes in the blood of calves of the experimental group increased by 18.8 and 26.7%, respectively, compared with 4.8 and 6.6% in the control group; the relative and absolute content of blood B-lymphocytes of calves of the experimental group increased by 21.5 and 97.6%, respectively, compared with 6.2 and 19.0% in the control group. A significant increase in the functional activity of blood neutrophils in calves of the experimental group was established in both induced and spontaneous tests. In calves of the experimental group, protective antibodies titers to PG-3, IRT and VD-BS viruses were observed on day 21 after vaccination; in the control group, a comparable level of antibodies was achieved on day 21 after revaccination.

Example 2. The comparative effectiveness of the prevention of respiratory diseases of calves by the claimed and known methods was determined on clinically healthy calves of 60 days of age. According to the principle of analogues, 4 groups of calves were formed. The calves of the three groups were immunized intramuscularly in the middle third of the neck with two injections with an interval of 21 days between injections of the emulsion inactivated vaccine against parainfluenza-3, infectious rhinotracheitis and viral diarrhea, a mucosal disease at a dose of 2.0 ml. For 15 days, the calves of the first and second groups were additionally administered orally 20-30 minutes before feeding with alcoholic tincture from a crushed plant mixture of equal amounts of grass and purple coneflowers, common harmonica grass, small-leaved linden flowers and licorice roots in the form of 7% an aqueous solution prepared in boiled chilled water at a dose of 2.0 ml / kg body weight of the animal and 70% alcohol tincture of echinacea purpurea in the form of a 7% aqueous solution at a dose of 2.0 ml / kg body weight, respectively. The calves of the fourth group were intramuscularly injected with hyperimmune serum containing specific antibodies to PG-3 and RTI viruses in titers of at least 1: 1280 and 1: 256, respectively, at a dose of 1.0 ml / kg live weight three times with an interval of 10 days. Efficiency was evaluated by the main clinical indicators. The results are presented in table 3.

Example 3. The comparative effectiveness of the prevention of respiratory diseases of calves by the claimed and known methods was determined on clinically healthy calves of 60 days of age. According to the principle of analogues, 4 groups of calves were formed. The calves of the three groups were immunized intramuscularly in the middle third of the neck with two injections with an interval of 21 days between injections of live attenuated vaccine against parainfluenza-3, infectious rhinotracheitis and viral diarrhea - TRIVAC mucosal disease (VIEV) at a dose of 2.0 ml. For 15 days, the calves of the first and second groups were additionally administered orally 20-30 minutes before feeding with alcoholic tincture from a crushed plant mixture of equal amounts of grass and purple coneflowers, common harmonica grass, small-leaved linden flowers and licorice roots in the form of 7% an aqueous solution prepared in boiled chilled water at a dose of 2.0 ml / kg body weight of the animal and 70% alcohol tincture of echinacea purpurea in the form of a 7% aqueous solution at a dose of 2.0 ml / kg body weight, respectively. The calves of the fourth group were intramuscularly injected with hyperimmune serum containing specific antibodies to PG-3 and RTI viruses in titers of at least 1: 1280 and 1: 256, respectively, at a dose of 1.0 ml / kg live weight three times with an interval of 10 days. Efficiency was evaluated by the main clinical indicators. The results are presented in table 4.

As can be seen from tables 3 and 4, the claimed method provided the optimal preventive effect, which was expressed in reducing the number of diseased calves and an easier form of the disease.

A decrease in the incidence rate indicated an increase in resistance, which indicated an increase in the adaptive level. After recovery, the calves of the experimental groups developed well, their reactions to external stimuli were restored, they were mobile and active.

The obtained clinical data confirmed the achievement of the claimed technical result - an increase in prophylactic efficacy against the background of natural virus carrying by activation of protective mechanisms in all recorded parameters. So, an increase in the levels of immunoglobulins G and M indicates a stimulation of humoral immunity, and an increase in the functional activity of T and B lymphocyte cells increases the tension and duration of immunity.

Sources of information taken into account:

1. Urban V.P., Naimanov I.L. Diseases of young animals in industrial livestock. - M .: Kolos, 1984. - S.192-196.

2. RF patent No. 2111011 C1, A61K 39/295, A61K 39/155, A61K 39/265.

3. RF patent No. 2378014 C1, A61K 39/295, C12N 7/00, A61P 31/12.

4. Korovina N.A., Zaplatnikov A.L. Often sick children and the modern possibilities of immunoprophylaxis of acute respiratory infections // Difficult patient. Pediatrics. - 2006. - No. 9. - S. 29-32.

5. Zaplatnikov A.L. Immunoprophylaxis and immunotherapy of acute respiratory infections // Attending physician. - 2006. - No. 9.- S.50-56.

6. Gwaetney JMJr. Rhinoviruses: in Evans AS, Kalow RA Viral infections of human epidemiology and control, 4 ^th ed. New York: Plenum, 1997.

Claims (1)

A method for immunological prophylaxis of respiratory diseases of calves, comprising immunization with an associated vaccine against parainfluenza-3 viruses, infectious rhinotracheitis and viral diarrhea - mucosal diseases in combination with the use of non-specific prophylaxis of alcohol tincture of plant material, characterized in that a mixture of equal amounts of plant material is used herbs and inflorescences of Echinacea purpurea, common harmonica grass, small-leaved linden flowers and bare licorice roots, which is flooded with ethyl alcohol 70% concentration at a ratio of 1:10, kept in a dark place at a temperature of 18-20 ° C for 7 days, and tincture is prescribed inside as a 7-8% aqueous solution in a dose of 1.5-2.0 ml / kg of live weight of the animal for 15 days, and calves of 60 days of age are immunized.