RU2402207C1 - Method for enhancement of adaptation capabilities of cows - Google Patents

Abstract

FIELD: agriculture. ^ SUBSTANCE: invention relates to livestock farming. Method included adding 2.7-3.3% solution of low-molecular chitosan with molecular weight of 120 kDa, degree of deacetylation is 81%. Chitosan solution is administered once in dose of 2.0-3.0 ml/kg of body weight for two 5-6-day courses with an interval of 5-6 days. ^ EFFECT: method application improves indices that characterise adaptation capabilities of cow organism. ^ 3 tbl, 1 ex

Description

The invention relates to the physiology of animals and can be used to increase the adaptive capabilities of cows.

There are a large number of ways to increase the adaptive capabilities of the animal organism, involving the use of various means: vitadaptine, extract of eleutherococcus, lucevi, chitosan and others [1, 2, 3, 4, 5, 6, 7].

A known method of increasing the natural resistance of cows of foreign selection through the use of vitadaptine, which is a sterile form of wheat germ oil. Intramuscular administration of vitadaptin at a dose of 10 ml promotes activation of indicators of nonspecific resistance [7]. The disadvantage of this method is the need for intramuscular administration of the drug.

There is a method of increasing the adaptive capacity of the body to stress factors of various nature by applying the extract of Eleutherococcus in a dose of 50 ml per 100 kg of body weight once inside with food. The adaptogenic effect of eleutherococcus is considered to be the ability to cause in the body a state of not specifically increased resistance to various adverse factors. However, according to N.V. Lazarev [4], an increase in the general adaptive reaction of the body against the background of the use of Eleutherococcus reduces the body's resistance to the infectious process.

There is a method of increasing the functional capabilities of the body of cows by introducing lucevita - a bioactive drug with the ability to maintain the enzymatic status of the body and acting as an activator of intracellular processes, providing protection for the body under the action of adverse environmental factors. A 20% aqueous solution of lucevita was administered intramuscularly at a dose of 25 mg / kg of animal body weight 1 time per day for 3 days three times with an interval of 3 days. The effectiveness of the drug is manifested by a positive effect on metabolic processes, indicators of natural resistance [1]. The disadvantage of this method is the need for intramuscular administration of the drug.

The closest analogue is a method of increasing the adaptive capabilities of a cow's body by introducing into the diet chitosan acid-soluble with a molecular weight of 250 kDa (high molecular weight) and a degree of deacetylation of 88% at a dose of 2 ml / kg of animal body weight 2 times a day for 5-6 days. Re-use was carried out after 5-6 days. This drug was used on cows under complex technogenic pollution with heavy metals [3, 5, 6]. Chitosan had an indirect effect on energy metabolism, since, first of all, it showed the properties of a sorbent of heavy metal ions, and against this background, metabolic processes in the body of cows returned to normal.

The aim of the invention was to develop an effective way to increase the adaptive capabilities of cows.

This goal is achieved by the fact that cows kept under new environmental and economic conditions are injected with a 2.7-3.3% solution of acid-soluble chitosan with a molecular weight of 120 kDa (low molecular weight) and a degree of deacetylation of 81% at a dose of 2.0- 3.0 ml / kg of animal body weight once for two 5-6-day courses with an interval of 5-6 days.

A comparative analysis of the proposed solution with the prototype shows that the method differs from the known one in that low-molecular chitosan with a molecular weight of 120 kDa (kilodaltons) and a degree of deacetylation of 81% is used as a pharmacocorrector. The equimolar ratio of acetic acid, chitosan and water is 1: 3: 100.

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 the material, low molecular weight chitosan is used, obtained under the production conditions of Bioprogress CJSC VNIITIBP, Shchelkovo, Moscow Region. A 2.7-3.3% gel solution of chitosan is prepared in a 1% solution of acetic acid. Drinking is carried out in the morning before feeding at a dose of 2.0-3.0 ml / kg body weight for 5-6 days. After 5-6 days, the course is repeated according to the above scheme.

Chitosan, being a polysaccharide biopolymer of nature, is a fully or partially deacetylated chitin obtained from the crab shell of the king crab Paralitodes camtchatica Tilesius. By its chemical structure, chitosan is a copolymer of D-glucosamine and N-acetyl-D-glucosamine, combining the properties of a polysaccharide and polycation, which determines its immunostimulating effect, in which a macromolecule contributes to an increase in the general nonspecific resistance of the body to adverse environmental factors.

A large number of free amino groups in the chitosan molecule determines its properties to bind hydrogen ions and acquire an excess positive charge, so chitosan is an excellent cation exchanger.

The presence of a positive charge on the primary amino group of chitosan in slightly acidic (pH <6.2) aqueous solutions determines its main biological properties: biocompatibility, biodegradability, non-toxicity, high adsorption capacity, ability to absorb toxic substances and form complexes with other biopolymers having a total negative charge.

The mechanism of action of chitosan as an adaptogen is as follows: when ingested under the influence of digestive enzymes, part of chitosan is broken down, absorbed into the blood and absorbed by the body in the form of low molecular weight compounds - aminoglucans, which can be effectively used as a source of the corresponding aminomonose for the continuous synthesis of mucoglycans that make up the composition proteoglycan complex, which, in turn, forms a complex with hyaluronic acid - a regulator of tissue permeability and, playing azhnuyu architectural role in the formation of the walls of blood vessels. Thus, the presence of amino groups in the macromolecule, the positive charge of the macroion, as well as the antiradical activity of the polymer are crucial in the implementation of the adaptogenic action of the polymer.

In Russia, the first works on chitin date back to 1933-1934. (P.P. Shorygin), however, research in this area was begun about 30 years ago. To date, interest in chitin-containing drugs has increased unprecedentedly. The more scientists learn about the properties of chitin and chitosan, the wider the scope of their practical use.

An analysis of the literature indicates the high therapeutic and prophylactic effectiveness of chitosan, which gives reason for a wider test and its introduction into veterinary practice, in particular, with the aim of increasing the adaptive capabilities of the cows organism to new environmental and economic conditions.

It is known that chitosan activates the enzymes synthesized in connective tissue cells, which are responsible for the transport of nutrients and metabolic products of cells in the matrix, thus activating lymphatic drainage and increasing the effectiveness of therapy in the endoecological rehabilitation system [2]. But so far it is not known which chitosan performs activation in the best way and what is the role of oligomeric chitosan with different molecular weight and degree of deacetylation.

From the literature available to us, the use of acid-soluble chitosan with a molecular mass of 120 kDa and a degree of substitution of 81% is not known to increase the adaptive capacity of the body of cows.

All of the above served as the basis for testing chitosan with a molecular weight of 120 kDa (low molecular weight) and a degree of deacetylation of 81% as a drug that increases the adaptive capacity of the body of cows.

Execution example. To implement the proposed method was put scientific and production experience on the basis of LLC "Yasnaya Polyany" Troitsky district of Chelyabinsk region. Imported cows of the Simmental breed of Austrian breeding (imported to Yasny Polyany LLC in April 2007), selected on the basis of analog pairs with a body weight of 500-550 kg, aged 4.5 years, were used as the studied objects. Cows were divided into 3 groups. Feeding and caring for animals were similar.

The first group was a control group, by biochemical indicators of which they judged the state of adaptation processes in the body of cows contained in new environmental and economic conditions.

The cows of the second experimental group received an oral 3% gel solution of low molecular weight chitosan with a degree of deacetylation of 81% at a dose of 2 ml / kg body weight for 5 days with a repeat of the course after 5 days off.

The cows of the third group were given high molecular weight chitosan with a molecular weight of 250 kDa and a degree of deacetylation of 88% at a dose of 2 ml / kg of animal body weight 2 times a day for 5 days. Repeated use was carried out after 5 days.

Blood was taken for research in the morning before feeding before the experiment, and then on 10, 30, 60 days from the moment the drug was given.

Based on the fact that the adaptation of animals to new conditions is carried out by changing the functions of organs and systems of the body that require the mobilization of metabolic processes, indicators of carbohydrate and fat metabolism and indicators characterizing the antioxidant system of the body, which play an important role in the development of compensatory and adaptive reactions of the body, were selected.

The results are presented in tables 1-3. The appointment of chitosan increased the level of glucose in the blood serum of cows of the experimental group by 10.18% on the 10th day of the experiment, in comparison with the control group, and by 13.58%, compared with the initial level (in the prototype - by 5.39 % and 10.0%, respectively). On the 30th day, there was a significant increase in glucose levels by 1.9 times, compared with the background value (in the prototype - 1.8 times). Due to its adaptogenic properties, chitosan has a positive effect on energy metabolism.

Due to the lack of energy substrates, as evidenced by the low concentration of glucose in the blood of the cows of the control group (background values), the animal organism cannot cover its needs due to aerobic glycolysis. This leads to a compensatory increase in anaerobic glycolysis, as evidenced by the tendency to increase the concentration of lactic acid in the blood of the control group of animals, in comparison with the experimental groups. As the data in table 1 show, the lactate content in the blood of the cows of the experimental group increased by 10.06% by the 10th day of the experiment, in comparison with the initial values (in the prototype - by 22.10%). From the 30th day, against the background of the use of low molecular weight chitosan, a decrease in this indicator was observed, amounting to 1.40 ± 0.03 μmol / L. By the 60th day of the experiment, the concentration of lactic acid was 1.32 ± 0.02 μmol / L, which is lower than the background level by 33.67% (in the prototype - by 24.73%).

Thus, the use of a drug with a low molecular weight accelerates the utilization of intermediate oxidation products that inhibit the production of ATP in mitochondria and gluconeogenesis reactions, which contributes to a more efficient involvement of lactic acid in energy supply processes, as a result of which its level in the blood decreases.

Table 1
Indicators of carbohydrate metabolism in the body of cows on the background of the use of low molecular weight and high molecular weight chitosan

Group Background Research Dates (days) 10 thirty 60 Glucose, mmol / L I 1.64 ± 0.04 1.67 ± 0.06 1.62 ± 0.03 1.69 ± 0.06 II 1.62 ± 0.03 1.84 ± 0.05 ^** 3.10 ± 0.08 *** 2.70 ± 0.05 ** III 1,60 ± 0,03 1.76 ± 0.04 2.85 ± 0.06 *** 2.51 ± 0.04 * PVC, μmol / L I 219.9 ± 3.56 223.6 ± 3.64 223.3 ± 2.79 221.3 ± 3.16 II 213.6 ± 3.24 220.9 ± 3.60 207.7 ± 2.59 ** 192.3 ± 2.36 ** III 210.5 ± 2.59 213.5 ± 2.86 ** 211.35 ± 2.23 197.7 ± 2.28 * Lactic acid, mmol / l I 2.05 ± 0.04 2.14 ± 0.03 2.35 ± 0.05 2,30 ± 0,05 II 1.99 ± 0.03 2.23 ± 0.05 1.40 ± 0.03 * 1.32 ± 0.02 ** III 1.90 ± 0.03 2.32 ± 0.04 1.56 ± 0.03 * 1.43 ± 0.04 * Note: * p <0.05 ** p <0.01 *** p <0.001

In the group of cows treated with chitosan with a molecular weight of 120 kDa, the content of pyruvic acid did not undergo significant changes, compared with the background values. It only slightly increased by the 10th day of the experiment to 220.9 ± 3.60 mmol / L, then it decreased by the 30th day and amounted to 207.7 ± 2.59 mmol / L. On the 60th day of the experiment, the concentration of PVC against the background of the use of low molecular weight chitosan decreased to 192.3 ± 2.36 μmol / L, which is lower than the background level by 9.97% (in the prototype - by 6.08%).

Against the background of the use of the crab polymer, there were also changes in lipid metabolism, namely, the administration of the drug (chitosan with a molecular weight of 120 kDa and a degree of deacetylation of 81%) contributed to an increase in the level of total lipids already on the 10th day of the experiment, which is 1.8 times above the initial level and 1.6 times relative to the control group (in the prototype - 1.7 and 1.5 times, respectively). On the 30th day of the experiment, the changes were as follows: the level of total lipids was 64.38% higher than the background value and 53.84% higher in comparison with the group of cows that received the main ration of the farm (in the prototype - 54.14% and 43.08%, respectively). By the end of the experiment (60th day), this value was slightly reduced. Against this background, the concentration of low density lipoproteins (β-lipoproteins) in the blood of cows receiving low molecular weight chitosan was unreliable.

table 2
Indicators of lipid metabolism in the body of cows on the background of the use of chitosan low molecular weight and high molecular weight

Group Background Research Dates (days) 10 thirty 60 Total lipids, g / l I 3.63 ± 0.10 4.11 ± 0.13 3.90 ± 0.39 4.56 ± 0.32 II 3.65 ± 0.08 6.60 ± 0.10 6.00 ± 0.35 ** 5.35 ± 0.14 * III 3.62 ± 0.06 6.07 ± 0.09 5.58 ± 0.26 * 4.97 ± 0.23 β-lipoproteins, g / l I 7.85 ± 0.10 8.07 ± 0.09 8.01 ± 0.23 8.16 ± 0.11 II 8.03 ± 0.12 7.95 ± 0.08 8.01 ± 0.11 8.08 ± 0.13 III 7.82 ± 0.10 7.98 ± 0.09 8.03 ± 0.10 8.20 ± 0.15 Cholesterol, mmol / L I 4.16 ± 0.13 3.74 ± 0.07 4.24 ± 0.12 3.62 ± 0.10 II 4.24 ± 0.11 3.63 ± 0.10 ** 2.40 ± 0.10 *** 2.18 ± 0.06 ** III 4.18 ± 0.14 3.69 ± 0.11 4.20 ± 0.13 2.33 ± 0.09 ** Note: * p <0.05 ** p <0.01 *** p <0.001

The opposite trend was observed in the level of free cholesterol in the blood serum of cows of the experimental group, which manifested itself in a decrease in the concentration of this indicator on the 30th day of the experiment by 1.8 times relative to the control group.

At the end of the experiment (the 60th day of the study), the cholesterol level in the blood serum of the cows of the experimental group was 2.18 ± 0.06 mmol / L, however, it remained above the standard values (in the prototype - 2.33 ± 0.09 mmol / L )

A low molecular weight chitosan had a significant effect on the indices of the antioxidant system of the body of cows (table 3).

Table 3
The dynamics of the content of ceruloplasmin and malondialdehyde in the body of cows with the use of chitosan of low molecular weight and high molecular weight

Group Background Research Dates (days) 10 thirty 60 Ceruloplasmin, g / l I 1.20 ± 0.03 1.21 ± 0.02 1.17 ± 0.02 1.19 ± 0.03 II 1.18 ± 0.02 1.12 ± 0.02 1.32 ± 0.05 ** 1.44 ± 0.05 *** III 1.15 ± 0.01 1.18 ± 0.04 1.26 ± 0.04 1.34 ± 0.05 MDA, mol / l I 3.18 ± 0.08 3.29 ± 0.09 3.27 ± 0.08 3.23 ± 0.04 II 3.15 ± 0.07 3.13 ± 0.08 2.65 ± 0.09 ** 2.59 ± 0.08 *** III 3.19 ± 0.06 3.20 ± 0.06 2.67 ± 0.12 * 2.63 ± 0.10 * Note: * p <0.05 ** p <0.01 *** p <0.001

There was an increase in the level of the main antioxidant of animal blood serum - ceruloplasmin, in comparison with background values by 11.86% and 22.03% (30th and 60th day of the experiment) (in the prototype - by 9.56% and 16.52 % respectively).

On the background of the use of low molecular weight chitosan, the content of malondialdehyde decreased to 2.59 ± 0.08 mol / L on the 60th day of observation (in the prototype, to 2.63 ± 0.10 mol / L).

Based on the results obtained, it can be concluded that in the body of cows that received low molecular weight chitosan, intensive carbohydrate oxidation occurs, as a result of which the energy necessary to ensure the constancy of the energy potential of the body of cows is released, because The main criteria for sustainable adaptive behavior are the energy aspects of resistance.

Thus, the advantage of the claimed method, in comparison with the prototype and other proposals in this area, allows us to recommend chitosan with a molecular mass of 120 kDa and a degree of deacetylation of 81% for use in veterinary practice to increase the adaptive capabilities of the body of cows.

Information sources

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Claims (1)

A method of increasing the adaptive capabilities of cows, including the introduction of a polysaccharide nature into the diet of a biopolymer, characterized in that the cows are given a 2.7-3.3% solution of low molecular weight chitosan with a molecular weight of 120 kDa and a degree of deacetylation of 81% at a dose of 2.0- 3.0 ml / kg of animal body weight once for two 5-6-day courses with an interval of 5-6 days.