RU2363483C1 - Way of prevention of secondary immunodeficiency at young stock of cattle - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention concerns veterinary science area. The way includes injection of a nonspecific immunoglobulin. In addition 70% spirituous tincture of the crushed vegetative admixture of pine buds, grass and inflorescences of coneflower, rhizomes and roots of horseheal, taken in the ratio 1:1:1. The spirituous tincture is obtained at a parity of raw materials and an extractor 1:5 and maintained at temperature 18-20°C within 7 days. Animals obtain the tincture in the form of a 7-8% aqueous solution in a dose of 1.0-1.5 ml/kg of live weight within 15 days with a 24 hours interval. The way allows stimulating humoral and cellular immunity effectively.

EFFECT: possibility to stimulate humoral and cellular immunity effectively.

4 tbl, 3 ex

Description

The invention relates to veterinary medicine, relates to a method for the correction of secondary immunodeficiencies and can be used as the main or additional component of the 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.

Immunodeficiencies - violations of the body's immune status due to a defect in one or more mechanisms of the immune response, are manifested by increased infectious morbidity. Secondary immunodeficiencies are acquired, develop under the influence of adverse environmental factors on the body. In the first days of life in newborn calves, physiological immunodeficiency is caused by the absence of immunoglobulins in the body, characterized by a deficiency of the B-system, against which gastrointestinal diseases develop, manifested by diarrhea (the first critical period), at the age of 20-40 days, immunodeficiency is associated with the expenditure of colostral immunoglobulins in case of insufficiency of one's own immunopoiesis, it is characterized by insufficiency of the phagocytic and T-systems, against the background of which diseases with respiratory damage develop (second second critical period).

Known methods for the correction of secondary immunodeficiencies using non-specific stimulants of immunity (synthetic, bacterial, plant, biological origin), differing in structure and mechanisms of action [1, 2, 3, 4, 5, 6, 7]. The problem of the use of immunotropic drugs is due to the high dynamism of the immune system, its multicomponent nature, the presence of a large number of direct and feedback connections, so the same drug, depending on the initial state of the immune system, dose and frequency of administration, and a number of other factors, can cause directly opposite effects.

As the closest analogue, we consider a method for the prevention of colostrum immunodeficiency, including the use of non-specific immunoglobulin, which is injected subcutaneously or intramuscularly on the first day of life and repeatedly with an interval of 14 days for 2 months [8].

The purpose of the invention is to increase the effectiveness of the prevention of immunodeficiency in calves.

This is achieved by using a complex herbal medicine against the background of passive immunization with non-specific immunoglobulin, and a complex medicine is proposed as a herbal medicine, which is a 70% alcohol tincture of a crushed plant mixture of ordinary pine buds, grass and purple inflorescences, rhizomes and roots of elecampane high, taken in a ratio of 1: 1: 1.

The ingredients of the complex drug used 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 act simultaneously on all parts of the immune system, as well as having a pronounced antimicrobial effect on pathogenic and conditionally pathogenic microflora.

Pine buds have disinfectant, phytoncide, vitamin and soothing properties, effectively prevent the spread of microbial and viral infections in the body.

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.

Rhizomes and roots of Elecampane high have anti-inflammatory, antimicrobial, bactericidal and antifungal properties.

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: essential oil, phenolic compounds, flavonoids, polysaccharides, 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, membrane tightening by lysis, 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.

Essential oils (contain all components) are distinguished by several types of activity (antimicrobial, anti-inflammatory, antispasmodic and expectorant), the details and degree of which varies. So, the antimicrobial effect has a wide spectrum and is non-specific. Its mechanism is complex and consists in the destruction of the cytoplasmic membrane of bacteria with subsequent violation of metabolism, aerobic respiration, and the synthesis process. Anti-inflammatory activity is manifested in the protection of cells from further damage, in weakening the exudative phase of the process, in leukocyte and macrophage infiltration, in enhancing cell proliferation. It is partly due to the antioxidant effect, as a result, lysosomal, mitochondrial and cytoplasmic membranes are stabilized, capillary permeability decreases, and the phagocytic activity of macrophages and leukocytes increases.

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 by the cell , as well as for water coming from the outside and contributing to lysis.

Vitamins (vitamin C - ordinary pine buds, echinacea purpurea; vitamin E - elecampane high, vitamin A - echinacea purpurea) are involved in all metabolic processes, increase the body's resistance to diseases, have anti-inflammatory effects, and contribute to detoxification of the body.

The trace element zinc (ordinary pine kidney, elecampane high) is involved in the body's immune processes, selenium (high elecampane, which concentrates selenium, ordinary pine bud) is a powerful antioxidant, and iron (purple coneflower) is the main structural component of blood hemoglobin and heme-containing enzymes.

All plants of the complex drug have an antimicrobial effect, and the herb and inflorescences of Echinacea purpurea have a virus-neutralizing property, pine buds prevent the spread of viral infection in the body, which eliminates (attenuates) the causes of the disease, anti-inflammatory (elecampane high, echinacea purpurea), expectorant (pine buds ), antispasmodic (elecampane high) effect, reduces pathogenetic changes, and immunomodulatory activity of Echinacea purpurea enhances the protective, comp sensory-adaptive mechanisms of the body.

The high efficiency of a multicomponent herbal product 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.

The proposed herbal remedy is prepared as follows.

The buds of Scots pine, grass and Echinacea purpurea inflorescences, rhizomes and roots of Elecampane high are crushed (in dry form) according to the State Pharmacopoeia and mixed in a ratio of 1: 1: 1. 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. The finished product is stored in a dark place at a temperature of 5-10 ° C. Shelf life - 2 years from the date of manufacture.

In accordance with the classification of toxicity of substances according to GOST 12.1.007-88, the agent can be rated as relatively safe, since when determining the parameters of acute toxicity in mice weighing 20.0 g, its 20% and 40% aqueous solutions showed LD _{50 of} more than 17.8 g / kg body weight, and with intrauterine administration - more than 3.0 g / kg body weight.

Example: 100 g of ordinary pine buds, 100 g of grass and Echinacea purpurea inflorescences and 100 g of rhizomes and roots of Elecampane high were ground, mixed, placed in a glass vessel, filled with 1.5 l of ethyl alcohol at 70% concentration and kept at a temperature of 18-20 ° With in a dark place for 7 days. The finished product was poured, the medicinal raw material was squeezed out and filtered.

A method for the prevention of immunodeficiency in calves with the use of a complex drug, which is a 7-8% aqueous solution of 70% alcohol tincture of a crushed plant mixture of common pine buds, grass and purple coneflower echinacea, rhizomes and roots of elecampane high, taken in a 1: 1 ratio : 1, obtained at a ratio of raw materials and extractor 1: 5 and kept at a temperature of 18-20 ° C for 7 days, against the background of passive immunization with non-specific immunoglobulin is as follows.

70% alcohol tincture from the crushed plant mixture of common pine buds, herbs and purple inflorescences of Echinacea purpurea and rhizomes and roots of Elecampane high in the form of a 7-8% aqueous solution prepared in boiled chilled water, is used orally in a dose of 1.0-1.5 ml / kg of animal body weight at intervals of 24 hours for 15 days in addition to intramuscular 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 change in immunobiological parameters depending on the parameters of the proposed method was determined in an experiment on clinically healthy calves 5-7 days of age. The calves selected by the analogue method were divided into six groups. The calves of the first five groups were intramuscularly 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, the calves of the first four groups were additionally administered orally a complex drug in the form of an aqueous solution of 7% concentration for 15 days with an interval of 24 hours in doses of 0.5; 1.0, 1.5 and 2.0 ml / kg of live weight of the animal. Calves of the sixth group of drugs were not received. Natural resistance was assessed by indicators of bactericidal activity of blood serum (determined by the method of O.V. Smirnova and T.A. Kuzmina, 1966), lysozyme activity of blood serum (determined by the method of V.G. Dorofeychuk, 1968) and the functional activity of blood neutrophils ( were determined in the reduction reaction of nitro-blue tetrazolium according to A. N. Mayansky et al., 1979), the immune status was determined by the number of T and B lymphocytes (determined by the Pierce method, 1962) and by the content of the main classes of immunoglobulins IgG, IgM (determined by the method of Mancini et al., 1965). The results are presented in table 1.

Table 1 The effect of the complex drug on immunobiological indicators and indicators of general non-specific resistance depending on the dose Indicators Groups of animals Experience The control 1st- n = 14 2nd — n = 15 3rd- n = 15 4th — n = 14 The dose of the complex drug, ml / g body weight Intramuscularly nonspecific immunoglobulin n = 14 No drugs were used n = 14 0.5 1,0 1,5 2.0 in addition to intramuscular injection of non-specific immunoglobulin T-lymphocytes,%

B-lymphocytes,%

Functional activity of blood neutrophils,% - spontaneous test -

- induced test

Immunoglobulins, mg / ml G

M

Lysozyme activity of blood serum,%

Bactericidal activity of blood serum,%

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

The obtained results confirmed the validity of the parameters of the proposed method. The use of 70% alcohol tincture of a crushed plant mixture of common pine buds, herbs and purple inflorescences, rhizomes and roots of elecampane high, taken in a ratio of 1: 1: 1, obtained with a ratio of raw material and extractor 1: 5 and aged at a temperature of 18-20 ° C for 7 days, in the form of a 7% aqueous solution at a dose of 1.0-1.5 ml / kg of live weight of the animal, provided a stable increase in immunobiological parameters in calves relative to animals of the group using non-specific immunoglobulin and the group without the use of preparations ata. Changing the values of this parameter of the method is impractical, since when lowering the immunobiological parameters are higher than in the group without the use of drugs, but lower than in the group using nonspecific immunoglobulin, and when they are increased, they remained at the level of the proposed values.

Example 2. The dynamics of changes in immunobiological parameters was studied in 2 groups (experimental and control) of clinically healthy calves of the economy, long-term dysfunctional in gastrointestinal diseases. The calves of the experimental 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 and an additional complex drug was used inside as an aqueous solution of 7% concentration at a dose of 1.0 ml / kg of animal weight for 15 days with an interval of 24 hours. Calves of the control group of drugs were not received. Immunobiological parameters were determined at different age periods and evaluated, as in example 1. The results are presented in table 2.

table 2 Age-related dynamics of changes in immunobiological parameters in clinically healthy calves Indicators Age, days one 10 twenty thirty T-lymphocytes,%

B-lymphocytes,%

Functional activity of blood neutrophils,% spontaneous test

induced test

Bactericidal activity,%

Lysozyme activity,%

Immunoglobulins,% IgG

IgM

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

As can be seen from table 2, in the daily age differences were practically absent. In the animals of the experimental group, by the age of 10 days, there was an increase in the main indicators of cellular and humoral immunity, which lasted until the age of 20 days, and in contrast to the animals of the control group, the positive trend continued until 30 days of age. Of great practical importance was the increase in the number of blood B-lymphocytes (by 53.2% in 10-, by 65.3% in 20- and by 50.8% in 20 days of age compared with the control) and immunoglobulins in the blood serum of experimental calves (IgG - by 17.0% at 10-, 39.7% at 20- and 40.2% at 30 days of age compared with control; IgM - by 36.4% at 10-, 55 , 6% at 20- and 36.4% at 30 days of age compared with the control), which testified to better absorption of colostrum immunoglobulins by newborns and an earlier start of the synthesis of their own immunoglobulins.

Example 3. The effectiveness of the method for the prevention of secondary immunodeficiencies in comparison with the known ones was studied under conditions of a farm that is stationary dysfunctional in gastrointestinal diseases of calves due to exposure 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 . 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. Bacteriological and virological studies have confirmed the etiological role of rota- and coronaviruses, as well as bacteria of the genera Escherichia and Proteus. Incidence - 87-95%, mortality - 25-30%. Clinically healthy calves of 5-7 days of age were selected according to the principle of analogues, which were divided into four groups of 15 animals each. The calves of the first group were intramuscularly 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 complex drug was used inside, which was an 8% aqueous solution of a 70% alcohol tincture of a ground plant mixture of ordinary pine kidneys, grass and inflorescences of Echinacea purpurea, rhizomes and roots of Elecampane high, taken in a ratio of 1: 1: 1, at a dose of 1.0 ml / kg of animal weight for 15 days with an interval of 24 hours (the proposed method). The calves of the second group were intramuscularly 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 at a dose of 10.0 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 3 and 4.

Table 3 Immunobiological parameters in calves after a course of prevention Indicators Prevention Method Intramuscularly nonspecific immunoglobulin and inside tincture
n = 15 Intramuscularly non-specific immunoglobulin
n = 15 Subcutaneous serum convalescents
n = 15 Without the use of drugs
n = 15 T-lymphocytes,%

B-lymphocytes,%

Functional activity of blood neutrophils according to 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 4 The comparative effectiveness of the proposed and known methods for the prevention of secondary immunodeficiencies Indicators Prevention Method Intramuscularly nonspecific immunoglobulin and inside tincture n = 15 Intramuscularly non-specific immunoglobulin
n = 15 subcutaneous serum of convalescents
n = 15 Without the use of drugs
n = 15 Sick, goals one 2 2 3 Form of the disease light one 2 2 2 heavy one Preventive efficacy,% 93.3 86.7 86.7 80.0 Duration of the disease, days four 5-6 6-6.5 7

It was found that the calves of the first group (the proposed method) had higher immunobiological parameters compared with calves of other groups (table 3). Of the 15 calves of the first group, one calf fell ill, and the disease was mild (clinical picture: depression, decreased appetite, feces liquid, beige, defecation rate up to 5 times a day) and was easily treated (on the 2nd day of treatment improvement in general condition: the appetite returned to normal, feces acquired a thicker consistency; recovery occurred on the 4th day). In the second and third groups of 15 calves, 2 calves fell ill. In both groups, one calf was ill in a severe form (clinical picture: depression, lack of appetite, pain due to intestinal spasm, feces liquid, beige, defecation frequency from 5 to 7 times a day). In calves of the second and third groups with a mild form of the disease, improvement in the general condition was noted on days 4.5 and 5.5, and clinical recovery on the 5th and 6th days, respectively, with a severe form of the disease, a slight improvement in the general condition (appetite improved, feces became mushy consistency, frequency of bowel movements - 5 times a day) was observed on 5.5 and 6 days, and clinical recovery - on 6 and 6.5 days, respectively (table 4).

The experimental material on the totality of indicators characterizing the immune status and the level of natural resistance confirms the stimulating effect on the body of the proposed method, which is expressed both in stimulation of the humoral (increase in bactericidal activity of blood serum and lysozyme activity of blood serum), and cell (increase in the number of T- and B-lymphocytes, phagocytic activity of blood neutrophils) immunity.

Information sources

1. Subbotin V.M., Subbotin S.G., Alexandrov I.D. Modern medicines in veterinary medicine. Rostov-on-Don: Phoenix, 2001, 600 p.

2. I.F. Gorlov et al. // 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. Bulletin VIEV, issue 66, M., 1988, p.8-9.

Claims (1)

A method for the prevention of secondary immunodeficiencies in young cattle, including an injection of nonspecific immunoglobulin, characterized in that an additional 70% alcohol tincture of the crushed plant mixture of ordinary pine buds, grass and purple coneflower inflorescences, rhizomes and roots of elecampane, taken in the ratio 1: 1: 1, obtained at a ratio of raw materials and extractor 1: 5 and kept at a temperature of 18-20 ° C for 7 days, in the form of a 7-8% aqueous solution in a dose of 1.0-1.5 ml / kg live weight alive within 15 days with an interval of 24 hours