RU2604135C1 - Method of immunodeficiency prevention in heifers during coming of physiological ageing - Google Patents

Abstract

FIELD: veterinary science.

SUBSTANCE: invention relates to veterinary science. Method involves use of immunomodulatory preparation during cattle growing, which is based on vegetal raw materials in form of water-alcohol infusion. Heifers of 15-16-month old during coming of physiological ageing daily for 30-60 days 20-30 minutes before feeding are fed orally 0.09-0.10 ml per 1 kg of animal's weight with aqueous-alcoholic infusion diluted 1-2 hours before use in 120-140 ml of boiled water at temperature of 34-40 °C for 300-350 kg of animal's weight. Infusion is prepared as follows: 2 parts of purple echinacea, 1 part of inula rootstock, 1 part of calendula flowers are mixed with 70 vol.% of rectified ethyl alcohol in ratio 1:5 and settled at temperature of 37-38 °C for 14-15 days. Then obtained infusion is brought to 30 % concentration with distilled water.

EFFECT: invention aims at preventing immunodeficiency in heifers.

1 cl, 2 tbl, 1 ex

Description

The invention relates to veterinary medicine, in particular veterinary pharmacology.

Modulators of the human immune system are known (see EPR, z. 0042064, class A61K 35/14, 1981), under which any substances are taken that can cause a direct or indirect reaction of an animal or person, part of an organism or substance taken from an organism , to antigens to which the body has previously been sensitized, or such a reaction is inherent in the function of the human or animal immune system. Modulators have a broad effect on the entire immune system. These modulators are isolated from the dialysate of leukocyte extract.

A disadvantage of the known technical solutions is the limitation of the action. In addition, the use of drugs from animal tissues to increase immunity is limited due to the possible presence of infectious proteins and the unpredictability of the effects of immunotropic drugs.

Known methods for increasing immunity, safety and productivity of animals through the use of agents containing DAFS-25 and various forms of carotenoids (carotene, lycopene) on an oil basis (patent No. 2423109, patent No. 2419420). However, these funds are primarily stimulants of productivity and safety, and increased immunity with their use is negligible.

The use of immunostimulating substances based on plant materials, such as Echinacea (Bauer R., Wagner N. Echinacea species as potential immunostimulator drugs. In: Wagner H., Farnsworth NR (eds.), Economic and Medicinal Plant Research. Vol. 5. New York: Akademic Press, 1991, pp. 253-351) for the prevention of immunodeficiency in the body.

A disadvantage of the known proposal is the limited effect, and since it contains only one echinacea as the main active ingredient.

The closest method of the same purpose to the claimed invention in terms of features is a method for the prevention of immunodeficiency in cows in the dry and postpartum periods, including the use of an immunostimulating drug based on plant materials, such as echinacea, characterized in that in the dry period the cows are given an aqueous-immunostimulant alcohol tincture of echinacea with silver ions, the daily dose of which is 20-25 ml per 500 kg of animal weight, and in the postpartum period, to enhance the effect of of the immunostimulant, cows are given a water-alcohol tincture of Echinacea grass, licorice and elecampane rhizomes with silver ions in the following ratio of plant materials - 2: 1: 1 in a daily dose of 20-25 ml, based on 500 kg of live weight. In the method, alcohol tinctures from echinacea, licorice and elecampane are prepared in 70% ethanol, diluted with water saturated with silver ions to 30% alcohol solution. (Patent No. 2182826 A61K 33/38, A61K 35/78. Publ. 27.05.2002 prototype).

The disadvantage of this method is its limited effect, in addition, the use of an aqueous solution of silver ions in it requires the use of an expensive device for its preparation, and the absence of carotenoids, flavonoids, and essential oils with phytoncidal properties reduces its effectiveness of the complex effect. In addition, the preparation contains raw materials that are difficult to obtain by cultivation, which complicates the preparation of the immunostimulant and increases its cost.

Known methods do not allow effective prophylaxis of immunodeficiency in heifers during the period of physiological maturation due to the limited effect, low complex effect on the immune system, the absence of important biologically active substances in its composition and the presence of expensive raw materials necessary for the manufacture of an immunostimulating drug.

The technical result is to increase the effectiveness of the prevention of immunodeficiency.

The technical result is achieved by the fact that in the method of preventing immunodeficiency in heifers during the onset of physiological maturation, the use of an immunomodulating preparation based on plant materials in the form of a hydroalcoholic tincture containing echinacea purpurea, an elecampane rhizome prepared on the basis of 70 when growing cattle % ethyl alcohol, according to the invention, to heifers 15-16 months of age during the onset of physiological maturation daily for 30-60 days for 20-30 m prenatal feeding is given orally at a dose of 0.09-0.10 ml per 1 kg of animal weight diluted 1-2 hours before use in 120-140 ml of boiled water at a temperature of 34-40 ° C per 300-350 kg of animal water -alcohol tincture, which is prepared as follows: 2 parts of purple coneflower, 1 part of elecampane rhizome, 1 part of calendula flowers is mixed with 70 vol. % ethyl rectified alcohol in a ratio of 1: 5 and kept at a temperature of 37-38 ° C for 14-15 days, the resulting tincture was adjusted to 30% concentration with distilled water.

The novelty of the proposed proposal lies in the fact that when growing heifers from 15-16 months of age during the onset of physiological maturation, a water-alcohol tincture of plant material is given orally: herb Echinacea purpurea, elecampane rhizome and medicinal calendula flowers.

In addition, the claimed method for the prevention of immunodeficiency in heifers during the onset of physiological maturation differs from the prototype modes of processing of plant materials, the scheme and dosage of the use of the resulting preparation.

These differences allow us to conclude that the claimed technical solutions meet the criterion of "novelty."

The features that distinguish the claimed technical solution from the prototype are aimed at achieving a technical result and have not been identified in the study of patent and scientific and technical literature in this and related fields of science and technology and, therefore, meet the criterion of "inventive step".

The production name of the herbal preparation obtained for the implementation of the indicated method for preventing immunodeficiency in heifers during the period of physiological maturation is the word formed from the words “calendula”, “elecampane”, “echinacea” included in the form of a water-alcohol tincture - “Kaledekhin”.

A method for the prevention of immunodeficiency in heifers during physiological maturation was carried out as follows.

Heifers 15-16 months of age during physiological maturity daily for 30-60 days 20-30 minutes before feeding are given orally at a dose of 0.09-0.10 ml per 1 kg of animal weight diluted 1-2 hours before use in 120-140 ml of boiled water at a temperature of 34-40 ° C per 300-350 kg of the animal’s weight, a water-alcohol tincture containing echinacea purpurea herb, elecampane rhizome, into which medicinal calendula flowers are introduced in the following ratio of plant material - 2: 1: 1, and insist for 14-15 days at a temperature of 37-38 ° C.

The mechanism of action used in the method of the drug is based on the chemical composition of biologically active substances that are part of the plant material used for its preparation.

Echinacea purpurea (Echinacea purpurea) in its composition contains the substance echinacin and hydroxycinnamic acids, which stimulate the body to produce its own interferon - an important protein that blocks the division of RNA and DNA viruses. Stimulates the activity of T cells, which are an important mediator in the body's immune system. It stimulates the adrenal cortex, increasing metabolic processes, while not causing its depletion. It also has antibacterial, antifungicidal and antiviral properties.

Elecampane high (Inula helenium L.) contains essential oil (up to 3%), consisting of gelenin, alanthol, proazulene and a mixture of sesquiterpenic lactones (alantolactone, isoalantolactone, dihydroalantolactone), polysaccharides - inulin (up to 40-45%), inulinine, pseudoinulin; traces of alkaloids, saponins, resins, gums, pigments, vitamin E, fatty oil, mucous membranes and bitter substances. Elecampane high has immunostimulating, general strengthening, secretolytic, enhancing the function of the sex glands (males and females), anti-inflammatory, antimicrobial, antihypoxic, diuretic, sedative, expectorant, analgesic, enveloping, astringent, reparative, sweating, sweating.

Calendula officinalis (Calendula officinalis L.) in flower baskets contains carotenoids and flavonoids (carotene, lycopene, violaxanthin, citraxanthin, rubixanthin, flavoxanthin, flavochrom). Calendula inflorescences also contain polysaccharides, polyphenols, resins (3.4%), mucus (2.5%), nitrogen-containing mucus (1.5%), organic acids (malic, ascorbic and traces of salicylic). In the aerial parts of the plant, up to 10% of the bitter substance of calendula was found, the photophytoncidal properties are due to the presence of essential oil. Calendula officinalis has the same properties as elecampane, as well as hypotensive, cardiotonic, anticoagulant, desensitizing.

The drug Kaledekhin based on echinacea purpurea, elecampane rhizome and calendula flowers does not give side effects, it is simple in production technology and method of use, in therapeutic doses it is not toxic.

For its manufacture, non-scarce, environmentally friendly raw materials are used, which provides economic benefits. The active principle of Kaledekhin is oxycinnamic acids, sesquiterpene lactones, inulin, carotenoids, flavonoids and a number of other compounds that make up the tincture obtained from the claimed herbs. These substances play an important role in the manifestation of the immunostimulating properties of the plant. "Kaledekhin" normalizes factors of nonspecific resistance of the body, the activity of immunocompetent cells, stimulates metabolic processes.

The drug has an immunostimulating, general strengthening, secretolytic, enhancing the function of the endocrine glands, anti-inflammatory, antimicrobial, antispasmodic, hypotensive, cardiotonic, antihypoxic, diuretic, sedative, anticoagulant, analgesic, sensitizing, sensitizing, sensitizing, sensitizing, sensitizing, sensitizing, sensitizing, sensitizing, sensitizing, sensitizing, sensitizing, sensitizing, sensitizing, sensitizing, sensitizing, sensitizing, antipyretic, sensitizing, sensitizing, sensitizing, sensitizing, sensitizing, sensitizing, sensitizing, sensitizing, sensitizing,

The drug Kaledekhin based on echinacea purpurea, elecampane rhizome and calendula flowers was used daily for 30-60 days. If the timing of the use of the drug on heifers was less than 30 days, this reduced its prophylactic effect on the animal’s body, since during this time the biologically active components did not stimulate the lymphatic system of the body (i.e., the activity of T and B lymphocytes did not increase) and the metabolism was not activated. Using the drug for more than 60 days did not increase its biological activity, but this method of preventing immunodeficiency became more expensive. In order to ensure the optimal effect of the use of the claimed drug, the time of its administration should be 45 days.

When using the drug on heifers, it must be given to the animal 20-30 minutes before feeding. If the drug is used earlier than 20 minutes before the feed intake, then this time did not ensure its absorption of biologically active substances before the feed intake, as a result, the effectiveness of the drug decreased. If the drug is used later than 30 minutes before the feed intake, the effectiveness of the drug also decreased, since the body of heifers did not have time to adapt and the assimilation of biologically active substances did not occur at the proper level. For optimal effect from the use of the inventive preparation, it is necessary to apply 25 minutes before the start of feeding.

The drug Kaledekhin must be used on heifers at a dose of 9-10 ml per 100 kg of heifer mass.

If the dosage of the drug was less than 9 ml per 100 kg of heifers, a longer period of use is required.

If the dosage of the drug was more than 10 ml per 100 kg of calf weight, then fattening begins earlier than 26 days, then this is economically disadvantageous, since the additive is overused and the cost of the resulting product increases.

If the duration of use of the drug on heifers was less than 30 days, then this reduced its prophylactic effect on the animal’s body, since during this time the biologically active components did not stimulate the lymphatic system of the body (i.e., the activity of T and B lymphocytes did not increase) and the metabolism was not activated. Using the drug for more than 60 days did not increase its biological activity, but this method of preventing immunodeficiency became more expensive. In order to ensure the optimal effect of the use of the claimed drug, the time of its administration should be 45 days.

An example of a specific implementation of the method for the prevention of immunodeficiency in heifers during the onset of physiological maturation

A tincture is preliminarily prepared based on the collection of purple coneflower herb, elecampane rhizomes and calendula flowers; raw materials were purchased from the North Caucasian AIA VILAR plant. Dried raw materials were crushed, rhizomes, stems, leaves - up to 3 mm, flowers - up to 5 mm, which were mixed in certain mass ratios: 2 parts of purple coneflower, 1 part of elecampane rhizome, 1 part of calendula flowers. The technological process for the preparation of preparations consists of the following stages: mixing the collection of purple coneflower, elecampane rhizome and calendula flowers with 70 vol. % ethyl rectified alcohol in a ratio of 1: 5. For this, 1 part of the raw material was taken, 5 parts of 70 vol. % ethanol (i.e., 1: 5 dilution), was stirred, placed in a dark glass dish, closed tightly and placed in a thermostat at 37 ° C for 14 days, periodically stirred. Then it was filtered through a paper filter, after pressing the raw materials well. The resulting tincture was adjusted to 30% concentration with distilled water, after which it was mixed, insisted for 24 hours for better diffusion. Thus, the phytopreparation Kaledekhin is ready for internal use by animals. During long-term storage (for three years), the drug did not change its properties (Table 1). In appearance, the phytopreparation Kaledekhin is a dark brown liquid with a specific herbal smell and taste. The drug Kaledekhin contains 1.0 ± 0.1% hydroxycinnamic acids, essential oils - 3.5%, polysaccharide inulin - 42%, vitamin E, ascorbic acid.

Heifers are given orally water-alcohol tincture "Kaledekhin" from 15-16 months of age for 30-60 days once a day for 20-30 minutes before feeding at a dose of 0.09-0.10 ml per 1 kg of animal weight, diluted with 120-140 ml of boiled water per 300-350 kg of animal weight.

The study of the immunomodulating properties of kaledechin was carried out in vitro with the peripheral blood of heifers. As a reference, a well-known immunomodulator of thymic origin, tactivin, was used.

In preliminary experiments, the optimal doses of tactivin were selected - 6.7 · 10 ^-4 ml during incubation for 30 minutes, kaledekhin - 3 · 10 ^-3 ml.

The effectiveness of the drugs was evaluated by the effect on the formation of rosette in lymphocytes and neutrophilic granulocytes. For this, rosette formation was carried out with the addition of drugs to whole blood.

The results of the study showed that tactivin at a dose of 6.7 · 10 ^-4 in 1 ml during incubation for 30 minutes had a stimulating effect on the number of E-ROL, a moderate increase in EAC-ROL in animals.

As a result of studies, it was found that the level of E-ROL (T-lymphocytes) in the absolute content is 4316.9 ± 114.5 in 1 mm ³ , which is 1.3 times higher than in the control group of animals (3342.5 ± 134 , 2 in 1 mm ³ ). A similar effect of the drug was observed in relation to EAC-ROL. So, the absolute values of the amount of EAC-ROL (B-lymphocytes) were 1.3 times higher than in the control (2403.0 ± 111.2 in 1 mm ³ ). Incubation with tactivin did not have a significant effect on the content of late E-RONs and led to a slight decrease in early E-RONs.

The incubation of cells with kaledekhin at a concentration of 3 · 10 ^-3 ml caused an increase in the absolute number of T-lymphocytes in the reactions of rosette formation by 1.4 times relative to that in the control group. The level of EAC-ROL (B-lymphocytes) also increased from 1907.0 ± 117.2 to 2406.7 ± 99.5 in 1 mm ³ , which is 1.3 times higher than the control. There was also a tendency towards an increase in the number of late E-RONs and EAC-RONs. Along with this, a suppressive effect was revealed in relation to early E-RON (Table 1).

Thus, the data obtained led to the conclusion that tactivin and kaledechin in the in vitro system have immunoprotective effects, but the phytopreparation showed a more pronounced effect. At the same time, it was noted that under the action of kaledekhin, more pronounced rosette formation occurred in T and B lymphocytes, in addition, the receptor apparatus in neutrophilic granulocytes was activated.

An analysis of the data obtained showed that taktivin and kaledekhin showed similar immunoprotective effects when they were incubated with immunocompetent cells. Moreover, the maximum stimulating effect on the number of lymphocytes forming rosettes with sheep erythrocytes (T-lymphocytes) and the C3 component of complement (B-lymphocytes) was noted during incubation of lymphocytes with kalendekhin, while taktivin had a more pronounced effect on the number of late E-RON and EAC-RON in cows. This position indicated a positive effect of the receptor apparatus of immunocompetent cells.

To study the immunobiological properties of tactivin and kaledekhin preparations in vivo for heifers from 15-16 months of age, the kalendekhin drug was administered orally for 30-60 days once a day for 20-30 minutes before feeding at a dose of 0.09-0.10 ml per 1 kg of animal weight, diluted to 130-140 ml of boiled water per 300-350 kg of animal weight, after which morphological, biochemical and immunological parameters of animal blood were examined. Tactivin was administered subcutaneously 0.02 ml / kg for 3 days.

Studies of morphological indicators showed that in animals treated with tactivin (experiment 1), the number of red blood cells increased by 9%, and kalendekhin (experiment 2) - by 15% in relation to the control group; white blood cells in the first experimental group - by 15%, and in the second experimental group - by 15%. At the same time, there was a decrease in the number of lymphocytes in the experimental groups (by 11% and 16%, respectively), due to an increase in segmented neutrophils (1.3 times), relative to the control group of animals (Table 2).

During biochemical studies of the blood serum of heifers, the following data were obtained: the total protein in animals of the control group was 71.0 g / l, in the first experimental group this indicator was higher than the control group by 22%, in the second experimental group - by 25%.

The blood glucose content in the control group was 5.36 mmol / L, in heifers of the first experimental group - 5.89 mmol / L, which is 9% higher than in the control, in the second experimental group - 6.92 mmol / L or 29% higher than in control.

The urea content in the control was 4.32 mmol / L, in the first experimental group - 3.08 mmol / L (29% lower relative to the control), in the second experimental group - 2.35 mmol / L (46% lower relative to the control ) The cholesterol content in all groups was approximately the same.

From the results obtained, it can be concluded that the use of tactivin and kaledekhin stimulated metabolic processes in the body and increased the natural resistance of animals.

The study of bacterial phagocytosis, that the percentage of actively phagocytic neutrophils in the control group of animals was 43%, in the first and second experimental groups - 50% and 52%, respectively, 16% and 21% higher than the control group; the absorption capacity of neutrophilic granulocytes in the control group was 2.54, and in the first experimental group - 2.67 (5% higher than in the control), in the second experimental group - 2.82 (11% higher than in the control) . The percentage of digestion was 67% in the control group, 72% - in the first experimental group (7% higher), 76% - in the second experimental group (13% higher than the control group).

When determining the content of T-lymphocytes in the first experimental group, an increase in their content by 65% was observed, in the second experimental group by 76%, relative to the control group. The number of B-lymphocytes increased in the first experimental group by 75%, and in the second experimental group - by 81%, relative to the control group of animals.

Therefore, from the results of the studies it can be concluded that the preparations of tactivin and kaledekhin had a stimulating effect on the metabolic processes and the immune system of animals, namely the production of red blood cells, hemoglobin, total protein, and increased the activity of T and B lymphocytes, which allows us to recommend kalendekhin for use in animal husbandry to increase the immunobiological reactivity of heifers during the preparation of physiological maturation.

Claims (1)

A method for the prevention of immunodeficiency in heifers during the onset of physiological maturation, including the use of an immunomodulating preparation based on plant materials in the form of a water-alcohol tincture containing purple echinacea grass, elecampane rhizome prepared on the basis of 70% ethyl alcohol, when raising cattle, that heifers 15-16 months of age during the onset of physiological maturation daily for 30-60 days 20-30 minutes before feeding are given orally at a dose of 0.09-0.10 ml per 1 kg of animal mass diluted 1-2 hours before use in 120-140 ml of boiled water at a temperature of 34-40 ° C for 300-350 kg of animal mass water-alcohol tincture, which is prepared as follows: 2 parts of Echinacea purpurea, 1 part of elecampane rhizome, 1 part of calendula flowers is mixed with 70 vol. % ethyl rectified alcohol in a ratio of 1: 5 and kept at a temperature of 37-38 ° C for 14-15 days, the resulting tincture was adjusted to 30% concentration with distilled water.