RU2371191C1 - Bioaactive stimulation agent and correction method applying agent for secondary immunodeficiency of calves - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention concerns veterinary medicine, bioactive stimulation agent and method of secondary immunodeficiency correction applying the agent. Method involves subcutaneous injection of non-specific immunoglobulin and additional oral application of alcohol infiltration of minced herbal mix including grass and inflorescence of Echinacea purpurea, coltsfoot, harmala shrub grass and common licorice roots at equal amount in the form of aqueous solution of 7-8% concentration in the dosage of 1.5-2.0 ml per kg of live weight for 15 days at 24 hour interval. Bioactive stimulation agent renders positive effect on all animal organs and systems, adjusts metabolic processes and enhances organisms resistance (activation of B- and T-systems, phagocytosis), allowing for correction of secondary immunodeficiency to physiological standard level.

EFFECT: natural origin, easy metabolic assimilation and excretion, high compatibility with other medicines, absent sensibilisation and induction of immunopathological reactions.

2 cl, 6 tbl, 3 ex

Description

The invention relates to veterinary medicine, relates to means of plant origin, having an immunostimulating effect, and can be used as the main or additional component of stimulation of the immune system of young farm animals, mainly calves, to increase the overall resistance and immunological reactivity to the action of various damaging agents.

The use of immunomodulating drugs is mainly aimed at the correction of secondary immunodeficiencies, which are manifested in frequent, recurring, difficult to treat infectious and inflammatory diseases of various localization. For young cattle, the first days of life and the start of skim milk collection (at 20 days of age) are critical immunological periods, which is associated with the physiological characteristics of the body, in particular, the immunodeficiency state of the calves before the first milking of colostrum, and at 20 days of age - with the expenditure of colostrum protective proteins in case of insufficiency of their own immunopoiesis, as well as stresses due to a change in feeding and a transition from individual to group content. Thus, our studies conducted in farms of the Nizhny Novgorod region, long-term dysfunctional in gastrointestinal and respiratory infections, revealed immunodeficiency in clinically healthy calves of both 3-5- and 20-day-old age (tables 1 and 2).

Table 1 The immune status of calves 3-5 days of age in a hospital Indicators Clinically healthy calves in a dysfunctional economy Clinically healthy calves from a prosperous farm T-lymphocytes,% 18.2 ± 0.8 19.8 ± 1.0 B-lymphocytes,% 3.1 ± 0.2 5.2 ± 0.3 Functional activity of blood neutrophils,%: spontaneous test 3.2 ± 0.2 3.8 ± 0.4 induced test 6.4 ± 0.4 8.4 ± 0.7 Bactericidal activity of blood serum,% 39.4 ± 3.0 52.1 ± 2.4 Lysozyme activity of blood serum,% 1.5 ± 0.2 2.5 ± 0.2 Immunoglobulins, ml G 11.9 ± 0.1 13.4 ± 0.2 M 1.1 ± 0.02 1.8 ± 0.05

table 2 The immune status of calves 20 days of age in a hospital Indicators Clinically healthy calves in a dysfunctional economy Clinically Healthy Calves T-lymphocytes,% 21.7 ± 1.0 32.9 ± 1.8 B-lymphocytes,% 5.7 ± 0.4 8.0 ± 0.6 Functional activity of blood neutrophils,%: - spontaneous test 4.8 ± 0.3 7.1 ± 0.5 - induced test 16.8 ± 1.4 28.4 ± 1.2 Bactericidal activity of blood serum,% 61.4 ± 1.2 86.5 ± 2.0 Lysozyme activity of blood serum,% 2.2 ± 0.2 3.6 ± 0.2 Immunoglobulins, mg / ml G 12.0 ± 0.3 18.0 ± 1.5 M 1.0 ± 0.01 1.4 ± 0.02

In conditions of farms that are not healthy due to calf diseases, clinically healthy young animals of 3-5 days of age showed immunodeficiency conditions, characterized by a decrease in the relative content of B-lymphocytes by 67.7%, levels of immunoglobulins G and M by 12.6 and 63.6% accordingly, the functional activity of blood neutrophils (in the spontaneous test - by 18.8%, in the induced test - by 31.3%), bactericidal and lysozyme activity of blood serum by 39.2 and 50.0%, respectively (table 1), and in clinically healthy young animals of 20 days of age - immu no deficiency states, characterized by a decrease in the relative content of B-lymphocytes by 40.4%, populations and subpopulations of T-lymphocytes by 51.6% with a violation of their ratio, the level of immunoglobulins G and M by 50 and 40%, respectively, and the functional activity of blood neutrophils (in spontaneous test - by 47.9%, in the induced test by 69%), bactericidal and lysozyme activity of blood serum by 40.9 and 63.6%, respectively.

A number of non-specific stimulants of immunity are known (synthetic, bacterial, plant origin), which differ in structure and mechanisms of action. Of the known herbal preparations, the immunostimulating properties are most studied in the preparations of Eleutherococcus, motherwort, preparations from pumpkin seeds, purple coneflower grass, safflower levzea. However, data on the immunotropic effect of these drugs differ in inconsistency / 1, 2, 3, 4, 5, 6 /.

Also known is a plant-based drug tetraimmunophyte - a phytoextraction drug from St. John's wort, hay leaf, rose hips and beet root crops, which has an immunostimulating effect on the humoral immune response / 7 /.

Closest to the claimed is a tool with an immunostimulating effect, which is an aqueous extract from the herbs of Hypericum perforatum, creeping thyme, peppermint and coltsfoot, taken in equal amounts / 8 /.

The technical result consists in the creation of a new drug with an extended spectrum of action, which has immunomodulating, anti-inflammatory and antimicrobial action, which provides stimulation of the body's defenses based on pharmacologically active substances from plants.

Obtaining this result is achieved by using tinctures from a crushed plant mixture of equal amounts of grass and inflorescences of purple coneflower, coltsfoot, common harmonica grass and licorice root, which is poured with ethyl alcohol at 70% concentration at a ratio of 1:10, kept in a dark place at a temperature of 15-20 ° C for 7 days, filter, squeezing the raw material, and filter.

The used ingredients of the drug are widely used in herbal medicine. The rationale for choosing the components of the composition was data on the pharmacological properties and chemical composition of the medicinal plant material. Each component contains biologically active substances that simultaneously activate intracellular metabolism and metabolism, and also have a pronounced antimicrobial effect on pathogenic and conditionally pathogenic microflora.

The grass and inflorescences of Echinacea purpurea have immunomodulating 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.

Grass and inflorescences of coltsfoot have pronounced bacteriostatic properties and antioxidant activity.

Garmala vulgaris grass has a pronounced antimicrobial activity, confirmed by pharmacological studies.

Licorice roots have antioxidant activity, hepatoprotective effect, contribute to the restoration of suppressed immune status.

The chemical composition of the drug 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, copper 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 / 8 /.

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 (contain 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 (licorice - up to 9.46%, coltsfoot - at least 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 - grass and inflorescences of coltsfoot, Echinacea purpurea; Vitamin E - licorice naked, Vitamin A - Echinacea purpurea) are involved in all metabolic processes, increase the body's resistance to disease, have anti-inflammatory effects, and contribute to detoxification of the body.

The trace element zinc (licorice - 0.33 μg / g, harmala vulgaris - 0.78 μg / g) is involved in the body in immune processes, selenium (licorice - 12.14 μg / g, harmala vulgaris - 1.0 μg / g, coltsfoot - 22.9 mcg / g) is a powerful antioxidant, iron (echinacea purpurea - 48 mg / 100 g, harmala vulgaris - 0.14 mg / g, naked licorice - 0.7 mg / g) - the main structural component of blood hemoglobin and heme-containing enzymes, as well as in combination with cobalt (licorice - 0.03 μg / g, coltsfoot - 4.0 μg / g) has a stimulating effect on the immunogenesis of the virus GOVERNMENTAL infections, copper (mother coltsfoot - 43.3 g / g, liquorice - 0.2 g / g) increases leukocyte functional activity, lysozyme and bactericidal activity of blood serum.

The high efficiency of the drug is achieved through a 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 tincture of the crushed plant mixture of herbs and purple coneflowers, coltsfoot, common harmonica grass and licorice roots is prepared in a 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 1-3 mm in size, 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 15-20 ° 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 purple coneflower, coltsfoot, common harmonica grass and licorice roots), 5.0 L of ethyl alcohol at a concentration of 70% were added, put in a dark place and kept at a temperature of 15-20 ° C for 7 days, after which it was filtered through the canvas, squeezing the raw materials, and filtered.

To correct secondary immunodeficiencies in calves against the background of nonspecific immunization, a biologically active stimulant is used, which is an alcohol tincture from a crushed plant mixture of grass and purple coneflower, coltsfoot, common harmonica grass and licorice roots taken in equal amounts.

A method for the correction of secondary immunodeficiencies in calves using a biologically active stimulant, which is an alcohol tincture from a crushed plant mixture of grass and purple echinacea, coltsfoot, common harmonica grass and licorice roots, taken in equal amounts, against the background of non-specific immunization in the following way.

70% alcohol tincture from a crushed plant mixture of herbs and purple coneflowers, coltsfoot, common harmala grass and licorice roots in the form of a 7-8% aqueous solution prepared in boiled chilled water is used orally at a dose of 1.5- 2.0 ml / kg of animal body weight at an interval of 24 hours for 15 days in addition to subcutaneous injection of non-specific immunoglobulin at a dose of 1.0 ml / kg of animal body weight three times with an interval of 5 days.

The essence of the method is illustrated by examples.

Example 1. The stimulating effect of the biologically active stimulant in accordance with the invention on the immunological status of newborn calves was studied in the farm, long-term disadvantageous diseases of calves (incidence of 60-70%, mortality - 20-30%). In conditions of inferiority (deficiency of macro- and micronutrients) feeding of mother cows (winter stall period) according to the principle of analogues, taking into account the clinical and physiological state, breed and body weight according to the principle of analogues, 2 groups of newborn calves with a live weight of 28-30 kg were formed for experience and control. The calves of the first group (experimental - 12 goals) were used inside a biologically active stimulant in the form of an aqueous solution of 7% concentration in a dose of 2.0 ml / kg of animal weight for 15 days with an interval of 24 hours. Calves of the control group (12 goals) of the drug were not received. The parameters of the immunological status of calves were studied on days 3, 10, 20 and 30 according to modern tests generally accepted in biology. The results are presented in table 3.

Table 3 Immunobiological blood counts of calves. Age, days Indicators 3 10 twenty thirty T - lymphocytes,%

B-lymphocytes,%

Bactericidal activity,%

Lysozyme activity,%

Immunoglobulins, mg / ml G

M

Note: in the numerator - indicators of experimental animals, in the denominator - control

When analyzing immunobiological parameters (table 3), experimental and control calves found that at the age of 3 days the differences were practically absent or were insignificant. In animals of the experimental group, by 10 days of age, there was an increase in the main indicators of cellular and humoral immunity, which lasted until 30 days of age. Of great practical importance was the increase in the number of immunoglobulins in the blood serum of experimental calves, which indicated better absorption of colostrum in newborns and an earlier start in the synthesis of their own immunoglobulins.

Example 2. The effectiveness of the method of correction of secondary immunodeficiencies using a biologically active stimulator in accordance with the invention against a background of subcutaneous injection of nonspecific immunoglobulin at a dose of 1.0 ml / kg of animal body weight was studied three times with an interval of 5 days in a farm that is stationary dysfunctional gastrointestinal intestinal diseases of calves due to the impact on the body of associations of viruses, pathogenic and conditionally pathogenic microflora against the background of violation of the conditions of feeding and feeding of pregnant Hur and calves. Acute gastrointestinal disorders in young animals were detected at 2-3 days of age. Patients noted a decrease in heart rate and respiration, lowering body temperature, lowering of the orbit of the eyes, cyanosis of the visible mucous membranes, disheveled and dull hair, profuse diarrhea. Incidence - 60-70%, mortality - 25-30%. According to the principle of analogues, clinically healthy calves of 1-3 days of age were selected, which were divided into four groups of 15 animals each - three experimental groups, one group - control. The calves of the first group were subcutaneously injected with a non-specific immunoglobulin at a dose of 1.0 ml / kg of animal body weight three times with an interval of 5 days and an additional biologically active stimulant was used, which was an alcohol tincture from a crushed plant mixture of grass and purple coneflowers, mother-and- stepmothers, herbs, harmonica vulgaris, and licorice roots, taken in equal amounts, in the form of an 8% aqueous solution at a dose of 1.5 ml / kg of animal weight for 15 days with an interval of 24 hours (the proposed method). The calves of the second group were subcutaneously injected with non-specific immunoglobulin at a dose of 1.0 ml / kg of animal body weight three times with an interval of 5 days (prototype method). The calves of the third group were injected subcutaneously with serum of convalescents in a dose of 20.0 ml for 9 days with an interval of 48 hours (basic method). Calves of the fourth group of drugs were not received. All animals were monitored for 30 days, recording all cases of the disease, the nature of their course and the effectiveness of the treatment. Natural resistance and immune status were evaluated as in example 1. The results are presented in tables 4 and 5.

Table 4A presents the comparative effectiveness of the proposed and known method for the correction of secondary immunodeficiencies.

As can be seen from table 4, 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.

Table 4 The effectiveness of the method of correction of secondary immunodeficiencies using a biologically active stimulant Method for the correction of secondary immunodeficiencies subcutaneous non-specific immunoglobulin subcutaneous serum of animals of convalescents without the use of drugs additionally inside tincture Indicators T-lymphocytes,%

B-lymphocytes,%

Bactericidal activity,%

Lysozyme activity,%

Immunoglobulins, mg / ml G

M

Note: in the numerator - indicators before the start of the experiment, in the denominator - 5 days after the end of the experiment

Table 4A The comparative effectiveness of the proposed and known methods for the correction of secondary immunodeficiencies Indicators Method for the correction of secondary immunodeficiencies subcutaneous non-specific immunoglobulin subcutaneous serum of animals of convalescents without the use of drugs additionally inside tincture Sick, goals one 2 2 3 The form
mild to severe illness one 2 2 2 1 Preventive efficacy,% 93.3 86.7 86.7 80.0 Duration of the disease, days four 5-6 6-6.5 7

Example 3. The effectiveness of the method of correction of secondary immunodeficiencies using a biologically active stimulant in accordance with the invention against a background of subcutaneous injection of nonspecific immunoglobulin in a dose of 1.0 ml was studied three times with an interval of 5 days in a farm that is stationary unfavorable for respiratory diseases of viral-bacterial etiology of calves with an incidence of up to 60-80%. The etiological role of infectious rhinotracheitis viruses (RTIs), parainfluenza-3 (PG-3) and viral diarrhea - mucosal diseases (VD-BS) was confirmed by the isolation of antiretroviral antibodies to these viruses in the serum of calves in 57.6, 72.4 and 75.5 % of cases, respectively. Bacterial microflora was represented by streptococci, salmonella and proteins. The immunological status was characterized by a 10-12% decrease in the relative content of B-lymphocytes, populations and subpopulations of T-lymphocytes with a violation of their ratios, the functional activity of neutrophils, a decrease in the level of immunoglobulins G and M, bactericidal and lysozyme activity of blood serum. Clinically healthy calves of 10-15 days of age were selected according to the principle of analogues, which were divided into four groups. Efficacy was evaluated by immunobiological and clinical indicators (tables 5 and 6).

Table 5 Immunobiological parameters in calves after a course of therapeutic and prophylactic effects for the correction of secondary immunodeficiencies Indicators The method of preventive treatment subcutaneous non-specific immunoglobulin subcutaneous blood serum of animals of convalescence without the use of drugs additionally inside tincture Functional activity of blood neutrophils by the HCT test,%: spontaneous

induced

Bactericidal activity,%

Lysozyme activity,%

Immunoglobulins, mg / ml: IgG

IgM

Note: in the numerator - indicators before the start of the experiment, in the denominator - 5 days after the end of the experiment

Table 6 The comparative effectiveness of the proposed and known methods for the correction of immunodeficiencies Indicators Method for the correction of secondary immunodeficiencies subcutaneous non-specific immunoglobulin subcutaneous serum of animals of convalescents
n = 16 without the use of drugs
n = 15 additionally inside tincture n = 16 n = 15 Ill, goal. one 2 2 3 Form of the disease, light one 2 2 2 heavy one Efficiency% 93.8 86.7 87.5 80.0 Duration of the disease, days - 3-4 5-6 6-7 7-8 Note: in the numerator - indicators before the start of the experiment, in the denominator - indicators after the end of the experiment

Correction of immunodeficiency states to the level of physiological norm (table 4) prevented the development of respiratory pathology in calves (table 5).

Our studies have shown that a biologically active stimulant, which is an alcohol tincture from a crushed plant mixture of grass and purple echinacea inflorescences, coltsfoot, common harmonica grass and licorice roots, taken in equal amounts, has immunostimulating activity, which can be used to correct secondary immunodeficiencies and stimulate natural resistance in infectious diseases pathogenetically associated with deficiency of the immune system. The effectiveness of the claimed biologically active stimulant is ensured by the presence in its composition of substances that increase the functional activity of the antioxidant and immune systems, in particular, macrophage cells. The tool is non-toxic, contains bioavailable useful biologically active substances, including, for example, essential oil, flavonoids, kartinoids, phytosterol, saponins, has ecological purity.

Information sources

1. A.D. Altunin et al. // Health, breeding and protection of small domestic animals: Materials 1 Intern. conf. - Ufa, 2000.

2. I.F. Gorlov and others // Veterinary medicine, 1996, No. 7.

3. A.A. Ivanovsky // Veterinarian, 2000, No. 2.

4. A.V. Korobko // Veterinary Medicine, 2000, No. 5.

5. V.P. Nikolayenko // Veterinary Medicine, 1990, No. 4.

6. Sh.B. Sadykov // Veterinary Medicine, 1990, No. 4.

7. L. A. Makaradze // Veterinary Medicine, 1999, No. 3.

8. RU 22501101 C1, 2005.

Claims (2)

1. A biologically active stimulant, which is an alcohol tincture of plant materials, characterized in that a mixture of equal amounts of grass and purple echinacea and coltsfoot inflorescences, common harmonica grass and licorice root are used as plant materials, and the plant material is poured with ethyl alcohol 70% concentration at a ratio of 1:10, kept in a dark place at a temperature of 15-20 ° C for 7 days, filtered, squeezing the raw material, and filtered. 2. A method for the correction of secondary immunodeficiencies in calves, comprising subcutaneous injection of nonspecific immunoglobulin, characterized in that the biologically active stimulant according to claim 1 is additionally used internally in the form of an aqueous solution of 7-8% concentration in a dose of 1.5-2.0 ml / kg live weight of the animal for 15 days with an interval of 24 hours