RU2597748C1 - Method of increasing resistance of calves to mixed viral-bacterial respiratory infections - Google Patents

Abstract

FIELD: veterinary science.

SUBSTANCE: invention refers to veterinary science and is intended to increase resistance of calves to mixed viral-bacterial respiratory infections. Clinically healthy calves aged 20-30 days are orally administered with fodder or water probiotic Vetom 1.1. Additionally, calves are treated with aerosol immunotropic preparation, containing 3.5 % aqueous solution of infusion, consisting of a mixture of common pine (Gemmae Pini), echinacea purpurea herbs and blossom clusters (Echinazea pyrpurea), inula rhizomes and roots (Rhizoma et radix Inulae), and xymedone hydrochloride in ratio of components of 4 g of xymedone hydrochloride per 100 ml of 3.5 % aqueous solution of infusion, in an amount of 4.0 ml/m³ of room, three times every 72 hours with 40-minute exposure. For producing infusion, a mixture of pine (Gemmae Pini), herbs and blossom clusters of echinacea purpurea (Echinazea pyrpurea), inula rhizomes and roots (Rhizoma et radix Inulae) in ratio 2:1: 1 is poured with ethyl alcohol with 70% concentration at a ratio of 1:5, held in a dark place at temperature 18-20 °C for 7 days, filtered, pressing raw material, filtered.

EFFECT: using disclosed method provides higher preventive effectiveness, increased cases light disease, which can be treated and faster treatment.

1 cl, 6 tbl, 3 ex

Description

The invention relates to veterinary medicine, relates to a method of increasing the resistance of calves to mixed viral and bacterial respiratory infections and can be used in a system of preventive measures to increase overall resistance, humoral and cellular immunity.

Massive respiratory diseases of young cattle are complex multifactorial syndromes that occur against the background of the interaction of various infectious agents, changes in immune status, physical and environmental factors, and manifest themselves as mixed viral and viral-bacterial infections, which are characterized by a particularly malignant course. The main role in the development of mixed respiratory infections of calves belongs to viruses (parainfluenza-3 viruses, infectious Torno-syncytial virus, etc.) and bacteria (primarily Pasteurella multocida, Mannheimia haemolytica, as well as Chlamydophila pecorum and Mycoplasma bovis). The respiratory infection causative virus viruses individually, but especially in associations, inhibit individual parts of the immune system, both cellular and humoral. So, when animals with viral diarrhea - a mucosal disease - are affected, the cellular immunity is mainly affected (the number and functional activity of lymphocytes decreases), the biosynthesis of antibodies is inhibited, the concentration of total protein in the blood serum is reduced, and diarrhea and infectious rhinotracheitis, diarrhea and parainfluenza are associated with an associated infection -3 decreases the biosynthesis of antibodies to these pathogens. In addition, the diarrhea virus - a disease of the mucous membrane of cattle - is reproduced in white blood cells - lymphocytes, monocytes, neutrophils, thereby causing leukopenia. The infectious rhinotracheitis virus reproduced in the calf's body inhibits the function of B-lymphocytes, reduces the activity of alveolar macrophages.

Deficiency of natural resistance and immunological reactivity due to insufficient morphological development and weakening of the functional activity of organs and tissues that form protective forces require adequate immunocorrective measures.

Currently, for the prevention and treatment of immunodeficiency, herbal preparations are increasingly being used. Their main advantages are versatility and gentle effects on the body and, as a result, good tolerance, the absence, as a rule, of side effects and complications even with prolonged use. Due to the combination of active components, plants are able to provide a multifaceted effect on the body (immunostimulating, anti-inflammatory, antimicrobial, antiviral, detoxification, etc.), which creates the conditions for a complex and multifunctional effect with a therapeutic and prophylactic effect on both the organ affected by the disease and the whole organism generally. So, leaves of coltsfoot (Tussilago farfara L.) contain mucus, tannins, ascorbic acid, carotene, bitter glycoside tussilgin, inulin, organic acids and other substances. Preparations from the leaves of the plant have a pronounced expectorant, disinfectant and anti-inflammatory effect, which depend on the presence of essential oils and mucous substances. The leaves of the plant are used in the form of infusion (1:10) inside as an expectorant and enveloping agent for diseases of the respiratory tract. Echinaceae purpurea (Echinaceae purpurea Moench) contains a unique complex of biologically active substances (polysaccharides, polyphenols and bioflavonoids with pronounced antioxidant activity), a protein balanced by essential amino acids, and serves as a accumulator of selenium, zinc and other micro and macro elements. The combination of all components determines the immuno- and biostimulating properties of Echinacea preparations [1].

The experience of traditional medicine suggests that in the treatment and prevention of many diseases it is better to use herbs, rather than individual plants. Medicinal preparations, which include plants that act directly on the immune system, have anti-inflammatory and antibacterial properties, as well as the ability to regulate the functions of other organs and systems, which in turn affect the body's immune response, have a complex normalizing effect on a living organism, increase its resistance, activate metabolism, due to the joint, often synergistic effect on the body of many substances that make up the fees. In addition, plants containing a complex of vitamins in an optimal combination for the body can be included in drug collections. Rationally selected combinations of medicinal plants potentiate the healing properties of the collection. Certain types of plant activity may not be sufficiently pronounced, but they are significantly enhanced by the rational combination of several plants [2]. Thus, the collection of medicinal herbs, including dioica nettle, large burdock, pharmacy chamomile, oregano, perforated St. John’s wort, wormwood, psyllium, coltsfoot and sowing oats, significantly increases the preventive and therapeutic efficacy in respiratory diseases of animals [3], and collection, including St. John's wort, creeping thyme, peppermint, coltsfoot, activation of the immune systems, in particular T and phagocytic, which aggravate the pathological process and the effectiveness of the treatment of respiratory diseases [4].

It is known from the prior art that probiotics can have a positive effect on factors of natural resistance and the immune status of young farm animals and birds [5]. Thus, the probiotic prolam (Lactobacillus delbrueckii subsp. Bulgaricus, Lactobacillus acidophilus 43c, Lactococcus lactis subsp. Lactis 574, Lactococcus lactis subsp. Lactis 1704-5, Bifidobacterium animalis 83) increases the absorption capacity of serum bacteriophobia and 6 Bacillus subtilis) - the bactericidal and lysozyme activity of blood serum [7], ocarin (representatives of the normal microflora of a healthy person, mainly Escherichia and staphylococcus) - the level of total immunoglobulins, bactericidal and lysozyme activity of blood serum [8], Bacinil (based on metabolic products ma spore-forming bacteria - Bacillus subtilis) - the number of Τ- and B-lymphocytes [9]. The most promising are probiotics based on microorganisms with desired properties obtained by genetic engineering, for example, the Vetom 1.1 probiotic containing Bacillus subtilis bacteria VKPM B-10641, which are highly resistant to digestive juices and enzymes of the gastrointestinal tract and are capable of rapid settlement. In the intestines of animals, bacterial spores are transformed into vegetative forms and secrete human interferon α-2, leukocyte substances, antibiotic-like substances, enzymes, and other biologically active substances. 1 g of the preparation contains 1 × 10 ⁶ CFU (colony forming units) of live microbial cells of the bacterial strain Bacillus subtilis VKPM B-10641. Vetom 1.1 is used for the prevention and treatment of gastrointestinal diseases with the syndrome of diarrhea, bacterial and viral infections (salmonellosis, coccidiosis, colibacteriosis, dysentery, company and parvovirus enteritis, influenza, parainfluenza, rhinotracheitis, hepatitis, carnivore plague, etc.) as well as for the correction of immunodeficiency states, for the treatment and prevention of dysbacteriosis disease in calves, piglets, carnivores, dogs, birds, growth stimulation and development of young animals [10]. It was found that the use of Vetom 1.1 for the treatment of viral and bacterial gastrointestinal diseases of calves with the symptom complex of diarrhea stimulated cellular and humoral immunity factors (the number of T-lymphocytes, the content of α- and β-globulins, bactericidal and lysozyme activity increased in blood serum) [11 ].

The purpose of the invention is a biocompatible, harmless and environmentally friendly way to increase the resistance of calves to mixed viral-bacterial respiratory infections, allowing it to be used for the prevention and treatment of associated infections that occur with severe clinical manifestations; with atypically manifest forms of the disease, the diagnosis of which is difficult; in combination with recommended treatment regimens with specific antiviral and bacterial drugs.

This goal is achieved by using a complex of drugs that act on various immunosuppressive factors, complementing each other and enhancing the effect, including the probiotic Vetom 1.1 and an immunotropic drug containing a 3.5% aqueous solution of alcohol tincture, consisting of a mixture of pine kidneys (Gemmae Pini), herbs and inflorescences of Echinacea purpurea (Echinazea pyrpurea), rhizomes and roots of elecampane (Rhizoma et radix Inulae) in a ratio of 2: 1: 1, and xymedon hydrochloride.

The method is as follows.

For clinically healthy calves 20-30 days of age, probiotic Vetom 1.1 is used orally with water or food at a dose of 50 mg / kg body weight once a day for 15 days and the calves are additionally treated with an aerosol immunotropic preparation containing a 3.5% aqueous tincture solution , consisting of a mixture of pine buds (Gemmae Pini), grass and inflorescences of Echinazea purpurea (Echinazea pyrpurea), rhizomes and roots of elecampane (Rhizoma et radix Inulae) in a ratio of 2: 1: 1, and Xymedon hydrochloride (4 g of Xymedon hydrochloride per 100 ml 3.5% aqueous solution of infusions), the rate of 4.0 ml / m ³ room, t ehkratno every 72 hours exposure at 40 minutes.

Aerosol treatment of calves is carried out in a specially equipped sealed chamber, equipped with sewage and supply and exhaust ventilation. To obtain highly dispersed aerosols, a SAF supersonic aerosol nozzle with a productivity of at least 200 ml / min and a dispersion of the generated aerosol from 5 to 25 μm are used. Processing is carried out at an air temperature of 14 ° C and a relative humidity of at least 70%). Aerosol stability is achieved by adding 10-20% sterile glycerol or 10% glucose in powder to the total volume of the working solution.

An immunotropic drug is prepared as follows.

The buds of Scots pine (Gemmue Pini), grass and Echinacea purpurea inflorescences (Ehinacea purpurea L.), rhizomes and roots of Elecampane high (Inula Neleium L.) are crushed (in dry form) according to the State Pharmacopoeia and mixed in a ratio of 2: 1: 1. The raw materials are placed in a glass vessel and moistened with ethyl alcohol at a concentration of 70% in a ratio of 1: 5, kept in a dark place at a temperature of 15-20 ° C for 7 days, filtered, squeezed out the raw materials, and filtered. In accordance with the classification of toxicity of substances according to GOST 12.1.007-88, the drug can be evaluated as relatively safe, because when determining the parameters of acute toxicity in mice weighing 20.0 g, its 20% and 40% aqueous solutions showed LD ₅₀ more than 17.8 g / kg body weight, and with intrauterine administration - more than 3.0 g / kg body weight. Before use, prepare a 3.5% aqueous solution using boiled and cooled to 37 ° C tap water, which is mixed with xymedon hydrochloride at the rate of 4 g per 100 ml of a 3.5% aqueous solution.

Used tincture ingredients 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 links of pathogenesis, the symptoms of inflammatory diseases of the respiratory organs, and also have a pronounced antimicrobial effect on pathogenic and conditionally pathogenic microflora.

The pine buds (Gemmue Pini) have a disinfectant, phytoncide, expectorant, vitamin and soothing properties, effectively prevent the spread of microbial and viral infections in the body.

The grass and inflorescences of Echinacea purpurea (Ehinacea purpurea L.) 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 neutrophils of blood and increase the synthesis of protective antibodies and endogenous.

Rhizomes and roots of high elecampane (Inula Neleium L.) have anti-inflammatory, antimicrobial, bactericidal, antifungal, hemostatic, expectorant properties.

Xymedon hydrochloride (1- (2-hydroxyethyl) -4,6-dimethyl-1,2-dihydropyrimidin-2-one hydrochloride) has reparative, immunostimulating and desensitizing properties, does not have toxicity, and irritates the skin and mucous membranes. In veterinary medicine, Xymedon hydrochloride was used in combination with mixoferon for associated calf bronchopneumonia [12], and in a mixture with allogeneic bovine serum for the prevention and treatment of gastrointestinal and respiratory diseases of calves [13].

The effectiveness of the method of increasing the resistance of calves to mixed viral and bacterial respiratory infections was determined in scientific and production experiments in the conditions of stationary farms in this calf pathology of the Nizhny Novgorod region with an incidence rate of 80-90%, mortality rate of 24%, where the etiological role of parainfluenza virus associations was established. 3 (PG-3), infectious rhinotracheitis (IRT), viral diarrhea - diseases of the mucous membranes (VD-BS), adenoviruses (AD), respiratory syncytial virus (PC) and bacterial microflora (pa Terrell, Salmonella, Proteus, streptococci, Escherichia). The immune status was assessed by the following indicators: the relative content of T-lymphocytes (in the reaction of spontaneous rosette formation according to the method of N.I. Blinov, 1980), the functional activity of blood neutrophils (in the reduction reaction of nitro-blue tetrazolium (HCT test) according to the method of M.E. Vicksman and AM Mayansky, 1979), lysozyme activity of blood serum (according to the method of B.G. Dorofeychuk, 1969), the level of immunoglobulins of individual isotypes (G and M) (according to the method of Manchini, 1964). The choice of immunological parameters is based on the fact that according to the results of studies, the immunodeficiency state in mixed viral-bacterial respiratory infections of calves is characterized by defects in the cellular and humoral parts of the immune system, phagocytosis and a decrease in natural resistance (table 1).

As can be seen from table 1, in clinically healthy calves immunodeficiency was noted, characterized by a decrease in the relative content of T-lymphocytes by 12.5%, the functional activity of blood neutrophils in spontaneous and induced tests by 17 and 13%, respectively, and lysozyme activity of blood serum by 21% and bactericidal activity of blood serum by 26.58% relative to the lower boundary of the physiological norm.

The essence of the method is illustrated by examples.

Example 1. The dose-dependent effect of an immunotropic drug on immunity parameters was determined on 5 groups of clinically healthy calves 20-30 days of age: 4 groups - experimental, 1 group - control. Calves of all groups were treated with Vetom 1.1 probiotic inside with water at a dose of 50 mg / kg body weight once a day for 15 days. The calves of the experimental groups were additionally treated with an aerosol of an immunotropic preparation containing a 3.5% aqueous solution of tincture, consisting of a mixture of pine buds (Gemmae Pini), grass and Echinazea pyrpurea inflorescences, rhizomes and elecampane roots (Rhizoma et radix Inulae) in the ratio 2: 1: 1, and xymedon hydrochloride (4 g of xymedon hydrochloride per 100 ml of a 3.5% aqueous tincture solution), three times with an interval of 72 hours with an exposure of 40 minutes at a rate of 2.0; 3.0; 4.0; 5.0 ml / m ^{3 of the} room. Calves of the control group were not subjected to additional processing. Blood sampling from animals for studies was carried out before treatments (background) and 7-10 days after the completion of the experiments. The results are presented in table 2.

As can be seen from table 2, the immunotropic drug had a stimulating effect on the initially reduced immunobiological parameters of clinically healthy calves. In all experimental groups, there was a significant increase in the relative content of blood T- and B-lymphocytes, the functional activity of blood neutrophils in spontaneous and induced tests, the level of immunoglobulins G and M, and lysozyme activity of blood serum. However, more pronounced positive changes characterizing the stimulating effect on the mechanisms of cellular and humoral links of immunity were found in calves of the third and fourth experimental groups (doses of immunotropic drug - 4.0 and 5.0 ml / m ³ ). The dose of the immunotropic drug 4.0 ml / m ^{3 was} chosen as the optimal one, since the dose increase did not provide a significant increase in immunobiological parameters.

Example 2. To confirm the feasibility of the claimed method, three groups of clinically healthy calves of 20-30 days of age were formed: 2 groups — experimental, 1 group — control. Calves of all groups were treated with Vetom 1.1 probiotic inside with food at a dose of 50 mg / kg body weight once a day for 15 days. The calves of the first experimental group were treated with an aerosol of an immunotropic preparation containing a 3.5% aqueous solution of tincture, consisting of a mixture of pine buds (Gemmae Pini), grass and purple Echinazea pyrpurea inflorescences, rhizomes and elecampane roots (Rhizoma et radix Inulae) in the ratio 2: 1: 1, and xymedon hydrochloride (4 g of xymedon hydrochloride per 100 ml of a 3.5% aqueous tincture solution), three times with an interval of 72 hours at an exposure time of 40 minutes at the rate of 4.0 ml / m ^{3 of the} room. The calves of the second experimental group were aerosolized with a 3.5% aqueous solution of tincture of Echinacea purpurea at the rate of 4.0 ml / m ³ room three times with an interval of 72 hours at an exposure time of 40 minutes. Blood sampling from animals for studies was carried out before treatments (background) and 7-10 days after the completion of the experiments. The results are presented in table 3.

As can be seen from table 3, 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. So, the relative content of blood T-lymphocytes in the first experimental group significantly increased by 40%, in the second - by 16%, in the control group, changes in this indicator were unreliable. The relative content of blood B-lymphocytes in calves of the first and second experimental groups, respectively, increased by 54 and 23%, in the control group, changes in this indicator were unreliable. The functional activity of blood neutrophils in a spontaneous test increased in the first experimental group by 25%, in the second experimental and control groups of animals, changes in this indicator were unreliable; in the induced test in the first, second, experimental groups, this indicator increased by 40 and 20%, respectively, compared with 18% in the control group. The lysozyme activity of blood serum significantly increased in only animals of the first experimental group by 77%. The content of immunoglobulins G significantly increased in the first experimental group by 31%, in the second experimental group - by 19%, in the control group - by 16%. The content of immunoglobulins M significantly increased in the first experimental group by 56%, in the second experimental and control groups, changes in this indicator were unreliable.

Example 3. To determine the effectiveness of the claimed method according to the principle of conditional analogues (of the same age, breed, live weight, physiological state), 5 groups of clinically healthy calves of 20-30 days of age were formed (table 4).

All animals during the experiment were conducted clinical observations. The results are presented in table 5.

As can be seen from table 5, from the total number of calves of the first experimental group (the method in accordance with the invention), respiratory disease was recorded in one animal, while in the remaining experimental groups were registered in 6.3 and 17 animals, respectively, and in 6 animals of the control group . Respiratory diseases have been clinically manifested by fever, congestive hyperemia, inflammation of the nasal mucous membranes, and the release of serous and then mucopurulent exudate. Inflammation of the mucous membrane of the larynx and trachea caused increased secretion in these areas, narrowing of the slit of the larynx and lumen of the trachea, blockage of the bronchioles, which made breathing more difficult and increased respiratory noise, and a cough appeared.

With a pathological autopsy, calves were diagnosed with catarrh and swelling of the mucous membrane of the nasal cavity, larynx, trachea, and the accumulation of serous or purulent exudate in the nasal passages and trachea. The conjunctiva is edematous, hyperemic, the lungs are enlarged, the lumens of the bronchi and alveoli are filled with foamy fluid or mucopurulent exudate. As noted above, non-specific factors of humoral immunity (lysozyme and bactericidal activity of blood serum, content of beta-lysines, interferon, etc.) play an important role in viral-bacterial respiratory infections, which indicate the state of the animal’s natural defense mechanisms and their interaction with pathogens of infections. The results indicate significant changes in this link of immune defense (table 6).

As can be seen from table 6, the trend of increasing indicators of humoral immunity was most clearly observed in the first experimental group. So, by the end of the experiment, the bactericidal activity of blood serum increased by 61.1% compared with 35.4% (second experimental group), 34.5% (third experimental group), 27.7% (fourth experimental group) and 56, 1% (control group) and serum lysozyme activity by 77.2% compared with 27.7% (second experimental group), 18.2% (third experimental group), 27% (fourth experimental group) and 27, 7% (control group).

Thus, the studies showed the possibility and high efficiency of the claimed method in the prevention of mixed viral and bacterial respiratory infections of calves.

Sources of information taken into account

1. Kolesnik, N.D. Immunostimulating properties of Echinacea purpurea / N.D. Kolesnik, S.A. Semenov, N.I. Ivanchenko // Zootechnics. - 2004. - No. 12. - S. 16-17.

2. Immunodeficiency states / Ed. V.S. Smirnova, I.S. Freidlin. - St. Petersburg: Tome, 2000 .-- 568 p.

3. Vildanov R.Kh. Medicinal herbs for respiratory pathology in calves / R.Kh. Vildanov, R.Kh. Vildanova // Veterinary medicine. - 2005. - No. 4. - S. 11-13.

4. RU 2250110 C1, 2005.

5. Spasskaya T.A. The effect of probiotics on indicators of resistance and the immune status of calves: Abstract. dis ... cand. biol. Sciences: 03.00.13 / Spasskaya Tatyana Arkadevna. - M., 1998 .-- 21 p.

6. Erina T.A. Intestinal microbiocenosis and immune status of newborn calves with different morphofunctional development and their correction: author. dis ... cand. Vet.science: 02/06/02 / Erina Tatyana Anatolyevna. - Voronezh, 2015 .-- 20 p.

7. N.E. Gorkovenko ¹ , Yu.A. Makarov ¹ , N.N. Shulga ¹ , N.P. Tarabukina. On the effect of concentrated blood serum and sahabactisubtil on the body of newborn calves. - Agricultural biology. - 2011. - No. 4. - S. 102-107.

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9. P.A. Krasochko, I.A. Krasochko, Yu.V. Sanzharovskaya. The effect of the cell-free probiotic preparation Bacinil on the immune system of calves in the treatment of respiratory diseases. - Veterinarian. - 2011. - No. 4. - S. 11-14.

10. Guidance on the use of Vetom 1.1 in veterinary medicine, approved. Veterinary Department of the Ministry of Agriculture of Russia 07/13/98, No. 13-5-2 / 1319, October 28, 1997

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Claims (1)

A method of increasing the resistance of calves to mixed viral-bacterial respiratory infections, including the use of probiotic Vetom 1.1, which is administered orally with water or food to clinically healthy calves 20-30 days old, characterized in that the calves are further treated with an aerosol immunotropic preparation containing 3.5% aqueous solution tincture consisting of a mixture of pine buds (Gemmae Pini), herbs and inflorescences of purple coneflower (Echinazea pyrpurea), rhizomes and roots of elecampane (Rhizoma et radix Inulae), and xymedon hydrochloride in a ratio of 4 g of hydrochloride xymedonide per 100 ml of a 3.5% aqueous solution of tincture, at the rate of 4.0 ml / m ³ room, three times with an interval of 72 hours at an exposure of 40 minutes, and to obtain a tincture a mixture of pine (Gemmae Pini), grass and purple coneflowers Echinacea purpurea (Echinazea pyrpurea), rhizomes and roots of elecampane (Rhizoma et radix Inulae) in a ratio of 2: 1: 1 pour ethanol at 70% concentration at a ratio of 1: 5, kept in a dark place at a temperature of 18-20 ° C for 7 days, filter, squeezing the raw material, filter.