RU2763842C1 - Method for increasing metabolism and nonspecific resistance in farm animals - Google Patents

Abstract

FIELD: veterinary medicine.

SUBSTANCE: invention relates to veterinary medicine and can be used as a method for increasing metabolism and nonspecific resistance in farm animals. The method involves feeding the probiotic Enzymsporin and consists in the fact that the diet includes microcapsulated Enzymsporin in combination with microcapsulated Dorogov’s antiseptic stimulant, fraction 2 (ASD-2f), respectively, in doses of 0.05 g per 1 kg of body weight and 0.05 g per 1 kg of body weight, which are fed once a day for 30 days every other day.

EFFECT: invention provides development of a new method that has a complex corrective effect on the metabolism and nonspecific resistance of the body of domestic animals.

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Description

The invention relates to veterinary medicine and can be used to correct metabolism and nonspecific resistance in farm animals.

Despite the development of modern animal husbandry, metabolic disorders and natural resistance to one degree or another occur in many productive animals. This leads to the development of new biologically active substances and methods of their application, which have a corrective effect on metabolism and non-specific protective factors in animals.

Currently, there are various ways to correct metabolism and nonspecific resistance in farm animals. Known drug for the correction of metabolic processes and resistance of the organism of animals, which contains organic acids (succinic, citric, ascorbic), the amino acid methionine and water for injection. The drug is administered intramuscularly at a dose of 2.0-10.0 ml. (Patent of the Russian Federation No. 2447886. - 2012, authors Evglevsky A.A. and others). The disadvantage of the proposed drug is a relatively low effect on some parts of the immune system. In this case, the drug is used in the form of injections, which increases the complexity of preventive and therapeutic treatments.

A known method of obtaining a drug for stimulating nonspecific resistance and metabolism in animals, which includes mixing succinic acid with ASDF-2 tissue preparation, sodium nucleinate and biological industry waste obtained after culturing quail fibroblasts on a primary culture medium, including MEM medium (Eagle's minimum medium ), synthetic medium 199, and bovine blood serum. The use of the obtained drug has a pronounced stimulating effect on metabolism, increases the natural resistance of the animal organism (RF Patent No. 2473340, 25.08.2011, authors O. B. Sein and others).

Also known is a drug for stimulating metabolism and non-specific resistance of animals, which includes sodium nucleinate, biological industry waste obtained by cultivating quail fibroblasts on a primary culture medium, including a synthetic medium MEM (Eagle's minimum medium), synthetic medium 199, blood serum of cattle and antiseptic stimulator Dorogov F-2 (ASD F-2). The use of the drug allows you to increase the metabolism, growth and resistance of the animal organism (RF Patent No. 2470648, December 27, 2012, authors Masalov V.N., Sein O.B. and others).

The disadvantage of the methods specified in patents No. 2473340 and No. 2470648 is that for their implementation it is necessary to have biological industry waste (waste in the production of vaccines), which is obtained only in biofactories.

There is a complex preparation for the correction of metabolic disorders in farm animals, which contains iron compounds with plant erythrodextrins, copper sulphate, manganese sulphate, cobalt sulphate, zinc sulphate, sodium selenite, potassium iodide, lactoalbumin enzymatic hydrolyzate, yeast extract, phenol and distilled water, at a certain ratio of components. The drug is used in the form of injections to prevent anemia in piglets and normalize metabolism in farm animals (RF Patent No. 2430720, 10.10.2011, authors Akhmadeev R.M. and others). The disadvantage of this drug is its multicomponent nature and use only in the form of injections.

Known drug and method for the prevention and correction of pathological conditions, the normalization of metabolism, increasing the resistance of the organism of agricultural and domestic animals. This drug includes a complex of L-amino acids and a complex of organic and inorganic substances. This drug is used by injection in the form of an aqueous solution for prophylactic purposes 2 times a week for a month at a dose of 1.5-2.0 ml per 10 kg of body weight and for therapeutic purposes 2 times a day at a dose of 3.0-5 0 ml per 10 kg of body weight for 3-5 days. The drug is used for intoxication due to poisoning with synthetic and (or) food poisons in a 10-fold therapeutic dose (Patent of the Eurasian Patent Office No. 015078, 04/29/2011, authors Khoroshikh E.M. and others). The disadvantage of this drug is its complexity and complexity of application.

There is a method for stimulating the growth and increasing the resistance of farm animals, when using which the animal is injected with an agent consisting of a lysate of peripheral blood leukocytes and stem and (or) progenitor cells of animals, as well as a conditioned medium obtained by their joint cultivation. The method provides the induction of an immune response and stimulation of the growth of farm animals by regulating the proliferation of immune cells and differentiation and determining the spectrum of synthesized cytokines (RF Patent No. 2495565, 10/20/2013, authors Gabriel I.N. et al.). The disadvantage of this method is the difficulty of obtaining the components that make up the stimulant.

There is a method for stimulating the resistance of the animal organism, characterized by the use of a drug from the herb St. In this case, the drug is used in combination with 7-10% glycerol for 40-50 minutes (RF Patent No. 2075975, 1997, authors V.T. Sidorov and others). The disadvantage of this method is the relatively low efficiency and the use of aerosol treatments, which significantly increases the cost of animal stimulation.

A known method of stimulating nonspecific resistance of animals, which involves the introduction to animals of a tissue preparation obtained from the organs of healthy animals (thyroid, parathyroid, thymus, pancreas; uterus, liver, ovaries, testes), with the addition of ASD F-2, formalin and bee honey ( RF patent No. 2337673. 2008, author Moiseev O.N.). However, to implement this method, biological material is required, which is obtained from animals only after slaughter. The technological process of manufacturing the drug is multi-stage and requires special equipment. At the same time, the proposed method allows to stimulate only non-specific resistance and does not have a pronounced effect on metabolism.

A known method of obtaining a drug to increase nonspecific resistance and immunogenesis of animals, characterized in that 90 wt. including 0.2-03% agar suspension is mixed with 2.5 wt. hours of purified polysaccharide complex concentrate with the addition of 3.5 wt.h. 2,3,5,6-tetrahydro-6-phenylimidazo-[2,1-|3]-thiazole hydrochloride and 0.2 wt. hours of formalin (RF Patent No. 2332214, August 27, 2008, authors Petryankin F.P. and others). The disadvantage of this method is the complexity of obtaining the drug, the scarcity of the components that make up its composition and relatively low efficiency. At the same time, this drug has a stimulating effect only on the nonspecific resistance of animals.

A method of obtaining a drug for increasing the resistance of newborn young animals of farm animals and a method for increasing resistance, which involves the use of a composition containing 1 part of a 3.5-4.0% ethanol extract of pollen as a source of trace elements, and 9 parts of nonspecific immunoglobulin, isolated from the blood serum of animals by treatment with polyethylene glycol (RF Patent No. 2570393, 2019, authors Ivanov A.A. et al.). The disadvantage of this method is the complexity of obtaining the components used in it, in particular, obtaining animal blood serum and immunoglobulins.

As a prototype, the probiotic preparation enzymsporin (LLC "Altbiotech", Russia. Manual on the use of the drug), which is a consortium of bacteria of the genus Bacillus with a bacteria content of at least 5⋅10 ⁹ CFU/g, was chosen.

The disadvantage of this drug and the method of its application is that when entering the stomach of animals, most (70-80%) of the probiotic bacteria Bacillus subtilis and Bacillus licheniformis die under the action of hydrochloric acid. Given the high percentage of death of probiotic bacteria in the acidic environment of the stomach, their doses when fed to animals are overestimated, which is economically unjustified. At the same time, enzymesporin has a positive effect on metabolic processes, but does not increase the nonspecific resistance of the organism.

The technical objective of the invention is to develop a new method that has a complex corrective effect on the metabolism and non-specific resistance of the organism of domestic animals.

The solution of the technical problem is achieved by the fact that microencapsulated probiotic enzymsporin and microencapsulated antiseptic - Dorogov's stimulator 2 fractions are additionally included in the diet of animals. At the same time, microencapsulated enzymes are obtained according to the method developed earlier by us (RF Patent for the invention No. 2689164, 2019, authors Trubnikov D.V., Sein O.B., Gorobets A.Yu., Trubnikova E.A.), the essence of which consists in the fact that 1.0 g of enzymesporin probiotic is dispersed in a suspension of 3.0 g of sodium alginate in 100 ml of distilled water with stirring 800-1000 rpm and precipitated with 100 ml of 50% acetone in the presence of 25 ml of 0.2 M calcium chloride solution with constant stirring for 15-20 min, and then the cured microcapsules are filtered on a Schott filter and dried at 30-35°C at a mass ratio of probiotic / sodium alginate 1:3.

Used in the claimed method, the second component - microencapsulated tissue preparation ASD-2 f - was also obtained according to a method previously developed by us (RF Patent for the invention No. 2543632. - 2015, author Sein O.B., Naumov M.M., Bykovskaya E.E., Krolevets A.A., Bogachev I.A.), using the non-solvent precipitation method, in which kappa-carrageenan is used as the shell of microcapsules, and ASD - 2 f.

To implement the proposed method, the animals are added to the diet microencapsulated enzymesporin at a dose of 0.05 g per 1 kg of body weight and microencapsulated ASD - 2 f at a dose of 0.05 g per 1 kg of body weight. Before use, these preparations are mixed with a small portion of compound feed and evenly introduced into the diet of animals. Microencapsulated preparations are fed to animals once a day every other day for 30 days.

A distinctive feature of the proposed method is the complex use of microencapsulated probiotic enzymsporin and tissue preparation ASD-2 f, which has a pronounced stimulating effect on metabolism and nonspecific body defense factors. The probiotic enzymsporin contains three strains of spore-forming bacteria of the genus Bacillus in equal proportions: Bacillus subtilis VKM B - 2998D (VKPM B - 314); Bacillus licheniformis VKM V - 2999D (VKPM V -8054); Bacillus subtilis VKM V - 3057D (VKPM V - 12079). The drug is a fine powder, highly soluble in water. Enzymsporin retains its properties after high temperature treatment for 20 minutes, does not contain GMOs. This probiotic is used in animal husbandry and veterinary medicine for the prevention and treatment of dysbacteriosis, increasing the body's natural resistance, normalizing the microbial balance in the digestive tract, stimulating growth, increasing the survival rate and growing healthy young animals.

Capsules from sodium alginate, which is a natural polysaccharide and which is obtained from seaweed, protect the probiotic bacteria that make up enzymes from destruction in the acidic environment of the stomach, which ensures their unhindered passage into the intestine, where, under the action of the alkaline environment of intestinal juice, the capsule shell is destroyed and probiotic bacteria enter the intestinal cavity. Reproducing in the intestines, probiotic bacteria synthesize and secrete biologically active substances (enzymes, amino acids, vitamins), which have a positive effect on the interior state of the intestines and metabolism in the animal body as a whole.

The second drug used in the method - microencapsulated ASD-2f, has a pronounced stimulating effect on biochemical processes, in particular, phosphorylation and protein synthesis, has immunostimulating effects, stimulates the activity of the reticuloendothelial and endocrine systems, increases the activity of tissue and digestive enzymes, accelerates the processes of regeneration of damaged tissues, has an antiseptic effect. The drug ASD - 2 f is widely used to stimulate the activity of the central and autonomic nervous system, increase the natural resistance of debilitated animals with diseases of the gastrointestinal tract, genitourinary system, skin diseases and metabolic disorders.

The microencapsulated enzymes and ASD-2 f used in the method in combination create a novel effect, in particular, they activate metabolism and nonspecific resistance, which has a positive effect on increasing the productivity and safety of animals.

Example 1. An experiment on pigs was carried out in the pig-breeding complex of APK Promagro LLC, Belgorod Region, Starooskolsky District.

The object of the research was 4-month-old Large White pigs, of which three groups of 10 heads each were formed in compliance with the principle of analogues.

Pigs of the 1st experimental group with the diet received microencapsulated enzymesporin at a dose of 0.05 g/kg of body weight (2.0 g per head) in combination with microencapsulated ASD-2f at a dose of 0.05 g/kg of body weight (2.0 g for 1 head). The preparations were fed once a day for 30 days every other day.

Pigs of the 2nd experimental group received microencapsulated enzymes at a dose of 2.0 g per head. The drug was fed with the diet once a day for 30 days every other day.

Pigs of the 3rd experimental group received non-microencapsulated enzymsporin according to the instructions for the drug (LLC "Altbiotech", Russia. Instructions for the use of enzymsporin) at a dose of 0.12 g/kg of body weight (5.0 g per head). The drug was fed once a day for 30 consecutive days.

Pigs of the 4th group were control and did not receive drugs.

Before feeding, the preparations were mixed with a small portion of compound feed and evenly introduced into the diet. The conditions of keeping the animals included in the experiment were the same. Before setting up for the experiment and at the end of the experiment, the absolute body weight was determined in all animals and blood was taken for laboratory analysis.

In the blood, general hematological parameters (ESR, hematocrit, erythrocytes, leukocytes, hemoglobin) were determined using conventional methods and an automatic hematological analyzer HumaCount 30TS (Germany).

The content of blood biochemical components (total protein, total calcium, inorganic phosphorus, glucose, total lipids, vitamin A, vitamin C, enzymatic activity of aminotransferases) was determined using Bio-La-Test reagent kits from Lachema (Czech Republic), Klini-Test" (Russia) and an automatic biochemical analyzer ILAB-650.

The content of total immunoglobulins in the blood of pigs was determined by the zinc sulfate method, the bactericidal activity of blood serum (BASK) was studied using a culture of Staphylococcus aureus. Serum lysozyme activity (LASK) was assessed using a daily culture of Micrococcus lisodecticus. The phagocytic activity of leukocytes (PAL) was determined by the reaction of phagocytosis with latex.

The experimental data obtained were subjected to biometric processing (P.F. Rokitsky, 1976). The degree of significance of differences was calculated using Student's t-test. The digital material was processed on a PC using the Microsoft Excel program.

The research results showed that during the experiment, the animals of all groups were healthy and had a good appetite.

In the course of a laboratory blood test, it was found that the general hematological parameters in piglets were within the physiological limits (Table 1). However, in piglets of the 1st experimental group, the content of erythrocytes and hemoglobin at the end of the experiment was significantly (p<0.05) higher compared to the background indicators and control animals, which indicates a higher "respiratory") function of their blood.

Studies of the biochemical components of blood showed that in pigs of the 1st experimental group, metabolic processes proceeded at a more intense level compared to animals of other (2,3,4) groups. As follows from the data presented in Table 2, the content of total protein, total calcium, glucose, vitamins A and C in the blood of piglets treated with microencapsulated preparations was significantly higher (p<0.05) than in control animals. This can be explained by the intensive exchange of these biochemical components and their entry into the blood from the intestinal tract.

The lower content of inorganic phosphorus in pigs of the 1st experimental group, apparently, is associated with its active participation as an energy and plastic material in biochemical processes that occur at a more intense level in animals that received microencapsulated preparations.

Significantly high content (p<0.05) of vitamins A and C in pigs of the 1st and 2nd experimental groups can be explained by its additional intake into the body as a result of active reproduction and increase in probionts of bacteria that produce a number of vitamins, including vitamins A and C At the same time, the improvement in the absorption capacity of the intestinal mucosa in pigs of the 1st and 2nd experimental groups also contributed to the intake of vitamins into the circulatory system of animals.

The enzymatic activity of aminotransferases (ACT, ALT) in the blood of pigs from the experimental groups had no significant differences (p>0.05) compared with control animals, which indicates the absence of a functional "overload" of the liver after the application of the developed method.

Studies have shown that the proposed method has a positive effect on the nonspecific protective properties of the organism (Table 3), in particular, in pigs of the 1st experimental group at the end of the experiment, the content of immunoglobulins G and M in the blood was significantly (p<0.05), higher bactericidal, lysozyme and phagocytic activity of blood in comparison with animals of the 2 experimental, 3 control groups.

Example 2. The experiment was carried out in the conditions of the educational and experimental farm of the Kursk KSAA named after I.I. Ivanova. The object of research was 4-month-old bulls of the Black-and-White breed.

4 groups of animals were formed, 10 heads each. Bulls of the 1st experimental group were fed microencapsulated enzymsporin at a dose of 0.05 g/kg of body weight in combination with microencapsulated ASD-2f at a dose of 0.05 g/kg of body weight in addition to the diet. The preparations were fed once a day for 30 days every other day.

Bulls of the 2nd experimental group were fed one microencapsulated enzyme at a dose of 0.05 g/kg of body weight. The drug was included in the diet once a day for 30 days every other day.

Bulls of the 3rd experimental group were fed non-microencapsulated enzymesporin according to the instructions for the drug (LLC "Altbiotech", Russia. Instructions for the use of enzymes) at a dose of 0.04 g/kg of body weight. The preparations were fed once a day for 30 consecutive days.

Bulls of the 4th group were control and received only the main diet.

Before feeding, the preparations were mixed with a small portion of compound feed and then evenly introduced into the diet. Experimental animals were kept in the same room.

As follows from the data presented in table 4, the bulls of the 1st experimental group had a higher hematocrit value, the content of erythrocytes, leukocytes and hemoglobin than in animals of other groups participating in the experiment. At the same time, the revealed differences in erythrocytes and leukocytes compared with control animals were significant (p<0.05).

Biochemical analysis showed (Table 5) that in the blood of bulls treated with microencapsulated enzymes in combination with ASD-2f, there was a higher content of total protein (p<0.05), glucose (p<0.05), total calcium ( p<0.05), vitamins A and C (p<0.05) in comparison with animals of the control group and background indicators.

At the same time, changes in the content of inorganic phosphorus, total lipids, as well as the enzymatic activity of ACT and ALT were unreliable (p>0.05).

The complex use of microencapsulated enzymes and ASD-2f had a pronounced effect on the nonspecific resistance of experimental animals. As follows from the data presented in table 6, all indicators reflecting the resistance of the organism in bulls of the 1st experimental group were significantly higher (p<0.05) compared to the control group. In bulls of the 2nd experimental group, which received microencapsulated enzymes, significant differences were recorded only in the content of JgG. In bulls of the 3rd experimental group, which received non-microencapsulated enzymes, the revealed differences were unreliable (p>0.05). At the same time, in animals of the 4th control group, the studied indicators were at a relatively low level.

Thus, the conducted studies indicate that the use of the developed method allows to increase the metabolism and nonspecific resistance in animals, it can be recommended for widespread use in the practice of animal husbandry and veterinary medicine.

Claims (1)

A method for increasing metabolism and non-specific resistance in farm animals, including feeding the probiotic enzyme enzyme, characterized in that the diet includes microencapsulated enzyme in combination with a microencapsulated antiseptic-stimulant Dorogov 2 fraction (ASD-2f), respectively, in doses of 0.05 g per 1 kg of body weight and 0.05 g per 1 kg of body weight, which are fed once a day for 30 days every other day.