RU2336693C1 - Method of natural cows resistance rising - Google Patents

Abstract

FIELD: agriculture, livestock farming.

SUBSTANCE: invention concerns veterinary science area. Cows are injected by 2.0-2.5% gel solution of chitosan acid-soluble with molecular weight 250 kDa and deacetylation range 88%, in dosage 2.0-3.0 ml/kg mass of a body 2 times a day during 6-7 days. After a week dosing schedule of chitosan is replicated. Additionally it is insulated elementary sulphur (with molecular weight 32) in dosage 3.0-3.5 g/100 kg of live weight in mixture with feed once a day during 25-30 days. Method positively effect on rates, describing functional status of humoral and cellular members of cows organism immune system.

EFFECT: receiving of rising method for natural cows resistance.

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Description

The invention relates to veterinary medicine and can be used to increase the natural resistance of cows. In territories contaminated by xenobiotics of chemical origin, including salts of heavy metals, the effect of depression of factors of natural resistance is observed in animals.

There are a large number of ways to increase the natural resistance of the organism of animals kept in conditions of technologically contaminated territory, involving the use of various means: cytodene, ligaverine, chitosan, additives "Ural", "Todikap-ideal" and others.

A known method of stimulation and correction of the depression of the immune system of young cattle, contained in conditions of significant technogenic pollution, by using cytodene, which is a dosage form of a biologically active immunoreactive protein belonging to the class of cytokines. A single intramuscular administration of cytodene in a single dose volume promotes the adhesive activity of neutrophils and an increase in the number of populations and subpopulations of lymphocytes (3). The disadvantage of this method is the need for injection of the drug.

A known method of increasing natural resistance by intramuscular injection of ligaverine in a dose of 5 ml daily for 5 days. Ligaverin, a complex of polymers isolated from plant materials, has an immunomodulatory effect aimed at stimulating the cellular component of the immune system, which is manifested by an increase in the number of leukocytes in the blood, the number of T-lymphocytes, and activation of phagocytosis (1). However, this method is quite time-consuming due to the impossibility of giving the drug inside with food.

A known method of stimulating the factors of natural resistance of the body of cows by oral administration of a feed additive "Ural" in a dose of 250.0 g per head daily, twice a day (5). Additive "Ural" is a natural product derived from peat, belonging to the group of humic preparations, containing a complex of biologically active substances. The positive effect of the drug is manifested by a positive effect on cellular immunity: the number of T and B lymphocytes increases, the phagocytic activity of neutrophils increases. However, this method is time-consuming in preparing the drug for feeding and requires the disposal of waste.

A known method of increasing the body's natural resistance by introducing "Todikap-ideal" in a portioned dose of 3 ml of liquid per calf per day for 10 days. "Todikap-ideal" is an extract of walnuts of milk-wax maturity in an organic solvent with the addition of propolis and pollen. The drug stimulates the immune system of animals, stabilizes the number of red blood cells, white blood cells, and increases the amount of total protein (2). However, data on the effect of the drug on indicators of the humoral link of the immune system are not available.

The closest analogue is a method of increasing the natural resistance of the body of cows by introducing into the diet chitosan, a natural biopolymer in the form of a 3% gel solution of 150 ml orally twice a day for 3 days a month for three months. The use of chitosan increases the phagocytic, lysozyme, bactericidal activity of the blood, and normalizes the number of T and B lymphocytes (4). This drug was used to treat endometritis in cows kept in a territory with a satisfactory environmental background. However, under the conditions of complex technogenic pollution of territories with heavy metals, the effect of the use of chitosan is insufficient.

The aim of the invention was to develop an effective way to increase the natural resistance of cows.

This goal is achieved by the fact that cows kept in ecologically difficult territories contaminated with heavy metals are injected with a 2.0-2.5% gel solution of acid-soluble chitosan with a molecular weight of 250 kDa and a degree of deacetylation of 88%, at a dose of 2.0 -3.0 ml / kg body weight 2 times a day for 6-7 days, with a weekly interval, the chitosan administration schedule is repeated, in addition to this, elemental sulfur with a molecular weight of 32 is introduced into the dose of 3.0-3.5 g / 100 kg of live weight mixed with food daily once a day for 25-30 days.

A comparative analysis of the proposed solution with the prototype shows that the method differs from the known one in that, in addition to chitosan, elemental sulfur is introduced in a dose of 3.0-3.5 g / 100 kg of live weight.

Thus, the claimed method meets the criterion of "novelty." Comparison of the proposed method not only with the prototype, but also with other solutions in this field allowed us to identify signs that distinguish the claimed solution from the known, which allows us to conclude that the criterion of "significant differences".

As a material, high molecular weight chitosan is used in combination with elemental sulfur. Apply chitosan obtained in the production environment of Bioprogress CJSC VNIITIBP of the city of Schelkovo, Moscow Region. A 2% gel solution of chitosan is prepared in a 1% solution of acetic acid. Drinking is carried out 2 times a day at a dose of 2.0-3.0 ml / kg body weight for 7 days. After 7 days, the administration is repeated according to the above scheme. Elemental sulfur is administered orally at a dose of 3.0-3.5 g / 100 kg of live weight mixed with food daily once a day for 25-30 days.

Chitosan is a derivative of chitin, a product of animal origin derived from shellfish. The aminopolysaccharide chitosan, which has a range of unique physicochemical properties and biological activity, is currently of interest to specialists in various fields and has prospects for widespread use, including in veterinary medicine.

In Russia, the first works on chitin date back to 1933-1934. (P.P. Shorygin), however, large-scale studies were begun about 30 years ago. Today, interest in these biopolymers has increased unprecedentedly. The more scientists learn about the properties of chitin and chitosan, the wider the scope of their practical use. And every year there are completely new and unexpected directions. The main areas of use of chitosan include medicine, agriculture, cosmetology and the food industry.

Chitosan is a polysaccharide biopolymer designed to protect living organisms from harmful effects. It is adapted to nature, so its use does not cause rejection by living organisms. Chitosan is a fully or partially deacetylated chitin, and the higher the degree of deacetylation, the more it contains reactive amino groups.

The available research results of domestic scientists, foreign experience in obtaining a wide range of chitin / chitosan-based drugs testify to the antiviral, antidote, antibacterial, antioxidant, anticoagulant properties, lipopolysaccharide-binding and immunoadjuvant activity of chitosan, which allows it to be used as an enterosorbent for effective fight against exo- and endotoxemia syndrome. The binding effect of chitosan depends on its molecular weight. High molecular weight chitosan (152-250 kDa) has a more pronounced adhesive effect than low molecular weight. The most pronounced sorption properties of chitosan are recorded in aqueous solutions of organic acids (formic, acetic). Upon dissolution of chitosan in an aqueous solution of acetic acid, a gel solution is formed.

Chitosan is a cationic polysaccharide with immunostimulating properties. All components of the immune system respond to polysaccharide immunomodulators, and their action is associated with macrophages and lymphocytes, the main components of the body's defense system. The mechanisms of the immunomodulatory effect of polysaccharides - glucans on the immune system are likely to be in their affinity with polysaccharides of the animal organism, which are mediators of cellular immunity.

Sulfur and its compounds are actively involved in many metabolic processes and perform very important functions in the body. Upon entering the body, part of the sulfur is absorbed by the bacteria of the digestive canal, and is converted into organic form, and is used to synthesize certain vitamins, hormones and sulfur-containing amino acids - cystine, cysteine, methionine.

The exchange of sulfur is closely related to the exchange and synthesis of a number of vitamins. At an optimal level of sulfur in the diet, the synthesis of sulfur-containing vitamins — thiamine, biotin, pantothenic acid, riboflavin, cyancobalomin, vitamin A — increases with a scar microflora. The optimal sulfur content in cattle diets should be 0.22% or 2.5 g per 1 kg of dry feed substances.

All of the above served as the basis for testing chitosan in combination with elemental sulfur as preparations that increase the natural resistance of the body of cows in technologically polluted areas.

Execution example. To implement the proposed method, experiments were conducted on the basis of the agricultural field "Niva" in the Stavropol district of the Samara region. The experiment was conducted on 32 cows of black-motley breed, selected according to the principle of analogues, with a live weight of 480-500 kg, aged 4-5 years, with a gestation period of 3.0-3.5 months. According to the principle of analogues, cows were divided into 4 groups. Feeding and caring for animals were similar. SHP "Niva" is located in an ecologically unfavorable zone of the Middle Volga region, where an abnormal content of heavy metals in the environment occurs.

The first group of cows served as a control; it was used to judge the state of natural resistance of animals kept in technologically polluted territories.

Cows of the second group were injected with chitosan in the form of a 2.0% helium solution in the optimal dose of 2.5 ml / kg body weight, established by preliminary studies, 2 times a day for 6-7 days with a course after 7 days of rest.

Cows of the third group were injected with chitosan in the form of a 2.0% gel solution at a dose of 2.5 ml / kg body weight 2 times a day for 6-7 days with a course after a week break and elemental sulfur at a dose of 3.2 g per 100 kg of live weight once a day mixed with food daily for 30 days.

Blood was taken for research in the morning before feeding before the experiment, and then after 20, 30, 60 days.

The results are presented in tables 1-4. The appointment of chitosan in combination with elemental sulfur reduced the number of stab neutrophils in the blood of cows by 42.50% (in the prototype by 35.58%). The level of segmented neutrophils is reduced by 25.66% (in the prototype by 15.90%). The content of eosinophils and monocytes increased to the values of the physiological norm and amounted to 6.02 ± 0.17 and 5.78 ± 0.13%, respectively (in the prototype 5.32 ± 0.10 4.73%).

1. Characterization of the quantitative composition of white blood cells in cows with the use of chitosan and chitosan in combination with elemental sulfur (X ± Sx, n = 10)

Indicator Group II 2nd III to experience after experience to experience after experience to experience after experience The total number of leukocytes, 10 ⁹ / l 7.05 ± 0.12 7.27 ± 0.14 7.38 ± 0.19 7.19 ± 0.10 7.13 ± 0.22 7.31 ± 0.17 Leukogram,% Basophils 1.5 ± 0.02 1,50 ± 0,02 1.00 ± 0.01 1.00 ± 0.03 1.20 ± 0.02 1.30 ± 0.01 Eosinophils 3.10 ± 0.02 3.04 ± 0.04 3.25 ± 0.04 5.32 ± 0.09 ** 3.30 ± 0.05 6.02 ± 0.17 ** Neutrophils: Young 1.00 ± 0.01 1.00 ± 0.02 1.00 ± 0.03 0.90 ± 0.04 1.00 ± 0.01 1.00 ± 0.03 stab 13.08 ± 0.24 13.24 ± 0.21 12.17 ± 0.46 7.84 ± 0.18 * 11.93 ± 0.29 6.86 ± 0.20 * segmented 49.03 ± 1.12 51.02 ± 2.18 49.98 ± 1.34 41.19 ± 1.16 * 49.34 ± 2.17 36.68 ± 1.96 * Lymphocytes 30.54 ± 0.92 28.60 ± 0.69 31.97 ± 1.12 39.02 ± 1.33 * 31.14 ± 1.46 42.36 ± 1.97 ** Monocytes 1.75 ± 0.03 1,60 ± 0,04 1.63 ± 0.04 4.73 ± 0.09 ** 1.79 ± 0.06 5.78 ± 0.13 ** Note: * - p <0.05; ** - p <0.001

Against the background of the use of chitosan in combination with elemental sulfur, a significant increase in the number of T and B lymphocytes was noted on the 30th day of the experiment, and by the 90th day the difference was 35.54 and 18.15% (in the prototype 16.21 and 9 , 08% respectively).

2. Quantification of peripheral blood lymphocyte populations in cows with chitosan combined with elemental sulfur (X ± Sx, n = 10)

Indicator Groups Background Research Days 1 20 days 30 days 60 days T lymphocytes I 31.37 ± 0.64 32.43 ± 1.34 31.29 ± 1.27 31.74 ± 2.08 II 33.24 ± 0.93 35.87 ± 1.31 37.58 ± 1.68 * 38.63 ± 1.32 ** III 31.68 ± 1.24 41.16 ± 2.14 ** 42.87 ± 1.73 ** 42.94 ± 2.89 ** B lymphocytes I 25.21 ± 1.03 27.83 ± 1.57 24.45 ± 1.29 24.93 ± 1.43 II 25.34 ± 0.68 26.72 ± 1.33 27.25 ± 1.24 27.64 ± 6.82 * III 24.63 ± 1.18 28.18 ± 1.02 * 28.92 ± 1.15 * 29.10 ± 1.42 * O-lymphocytes I 43.42 ± 1.98 39.74 ± 1.72 44.26 ± 1.69 43.90 ± 1.87 II 41.42 ± 1.12 37.40 ± 1.26 * 35.17 ± 1.67 ** 33.73 ± 1.49 ** III 43.68 ± 2.17 30.67 ± 2.16 28.21 ± 2.38 ** 27.96 ± 2.14 ** Note: * - p <0.05; ** - p <0.001

An increase in the number of phagocytic cells and their functional activity was established (Table 3). In the blood of animals of the experimental groups on the 60th day, the phagocytic activity of neutrophils increases by 12.34% (in the prototype by 5.20%).

The increase in the functional activity of phagocytes is indicated by the phagocytic number and phagocytic index, which increased during the use of chitosan in combination with elemental sulfur by 25.10-37.04% (in the prototype by 13.60-22.01%).

3. Evaluation of the phagocytic activity of cow blood neutrophils with the use of chitosan and chitosan in combination with elemental sulfur (X ± Sx, n = 10)

Indicators Group Background Study days The 20th 30th 60th Phagocytic activity,% I 52.27 ± 1.89 52.43 ± 1.43 52.14 ± 1.98 52.39 ± 1.82 II 52.72 ± 0.96 53.29 ± 1.52 56.24 ± 1.19 * 55.46 ± 0.84 * III 51.12 ± 1.27 53.84 ± 1.68 58.07 ± 1.64 * 57.43 ± 1.87 * Phagocytic index, cu I 3.60 ± 0.15 3.64 ± 0.17 3.60 ± 0.15 3.52 ± 0.16 II 3.68 ± 0.09 3.87 ± 0.12 4.32 ± 0.16 * 4.49 ± 0.16 * III 3.51 ± 0.15 4.09 ± 0.21 4.68 ± 0.19 * 4.81 ± 0.18 * Phagocytic number, cu I 2.64 ± 0.11 2.60 ± 0.10 2.67 ± 0.13 2.58 ± 0.12 II 2.72 ± 0.12 2.75 ± 0.13 2.86 ± 0.14 3.09 ± 0.12 III 2.59 ± 0.14 2.79 ± 0.15 3.07 ± 0.15 * 3.24 ± 0.16 * Note: * - p <0.05

Chitosan in combination with elemental sulfur have a significant effect on the indicators of the humoral link in the immune system of cows (Table 4).

4. Evaluation of some indicators of the humoral link of the immune system in cows with the use of chitosan and chitosan in combination with elemental sulfur (X ± Sx, n = 10)

Indicator Group BACKGROUND 30 days 60 days 90 days CEC,% I 70.83 ± 2.73 69.67 ± 2.82 69.81 ± 2.29 70.49 ± 3.06 II 70.08 ± 3.08 80.46 ± 3.15 * 87.53 ± 3.32 ** 92.15 ± 3.49 ** III 71.28 ± 2.14 74.18 ± 3.29 80.27 ± 2.31 84.18 ± 2.45 * Complement activity, conventional units I 6.89 ± 0.19 6.93 ± 0.27 7.04 ± 0.23 6.82 ± 0.14 II 7.21 ± 0.24 6.42 ± 0.20 5.89 ± 0.18 ** 5.32 ± 0.19 ** III 6.97 ± 0.20 6.69 ± 0.17 6.27 ± 0.14 * 6.12 ± 0.17 ** Lysozyme activity,% I 18.25 ± 0.56 17.86 ± 0.89 18.34 ± 0.45 18.22 ± 0.63 II 17.74 ± 0.68 20.37 ± 0.48 ** 22.19 ± 0.53 ** 22.87 ± 0.62 ** III 18.16 ± 0.36 19.12 ± 0.26 * 19.74 ± 0.39 * 19.26 ± 0.49 * Bactericidal activity,% I 48.97 ± 1.28 50.11 ± 1.19 48.29 ± 1.93 49.34 ± 1.34 eleven 50.12 ± 1.56 50.64 ± 1.68 50.18 ± 1.72 49.83 ± 1.49 III 50.73 ± 1.28 49.72 ± 1.42 49.58 ± 1.36 50.24 ± 1.37 Note: * - p <0.05; ** - p <0.01

There was a significant decrease in the level of the CEC in comparison with the baseline by 30.06% (in the prototype by 20.60).

The activity of the proteins included in the complement system decreases gradually and by the 60th day of the experiment, the difference with the background is 26.21% (in the prototype by 13.3%).

Against the background of the introduction of chitosan in combination with elemental sulfur already on the 30th day of observation, lysozyme activity increases, the maximum levels of lysozyme in the blood of cows reach the 60th day of the experiment, exceeding the background by 28.92% (in the prototype by 17.57 %).

Summarizing the established results, we can conclude: the introduction of chitosan in combination with elemental sulfur in the feed diet has a more pronounced positive effect on indicators characterizing the functional state of the humoral and cellular parts of the immune system of cows.

Thus, the advantage of the claimed method compared to the prototype and other proposals in this area allows us to recommend it for use in veterinary practice to increase the natural resistance of cows. In territories contaminated with xenobiotics, including heavy metals, animals develop immunosuppressive states and it is very important to use drugs that can increase the body's natural resistance.

Bibliography

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Claims (1)

A method of increasing the natural resistance of cows, including introducing into the diet of animals a natural polymer with an immunostimulating effect, characterized in that a 2.0-2.5% acid-soluble chitosan gel solution with a molecular weight of 250 kDa and a degree of deacetylation of 88% is used as an immunostimulant , at a dose of 2.0-3.0 ml / kg body weight 2 times a day for 6-7 days, after a week, the regimen of chitosan is repeated, in addition to this, elemental sulfur is introduced inside at a dose of 3.0-3.5 g / 100 kg live weight mixed with feed o twice a day for 25-30 days.