RU2289402C2 - Method for immunocorrection in cows - Google Patents

Abstract

FIELD: veterinary.

SUBSTANCE: claimed method includes administration of mixture of succinic and ascorbic acid in ratio of 1:1 in dose of 0.5-1.5 g/head/day per os for 20-25 days before calving and 20-25 days after calving.

EFFECT: method and agent for effective immunocorrection in cows.

2 cl, 4 tbl, 2 ex

Description

The invention relates to veterinary medicine, relates to a method of immunocorrection in cows, aimed at preventing (prophylaxis) various violations of the reproductive function of cows in the post-hotel period.

The weakening of the protective and adaptive mechanisms of the body is the main predisposing factor in the etiopathogenesis of postpartum diseases in cows.

The constancy of the cellular and humoral composition (homeostasis) of the body is controlled by the immune system. The work of the immune system is determined by immunocompetent cells, of which the main ones are the white blood cell in the entire diversity of its populations and subpopulations and tissue macrophages. All immunocompetent cells act in a complex manner (cell cooperation) in accordance with their function. So, phagocytes (monocytes, neutrophils and tissue macrophages) provide for the killing and elimination of a foreign agent, the development of auxiliary processes and the launch of many mechanisms of specific reactions that activate or suppress the immune response, T-lymphocytes - cellular, and macrophages and B-lymphocytes - humoral immunity. Under the influence of various kinds of immunosuppressants, insufficiency of immunity mechanisms (cellular or humoral or its individual links) develops [1, 2].

To compensate for immunological deficiency, various groups of drugs are used - non-specific immunostimulants (corpuscular immunostimulants: liposomes, latex, microcellulose, etc.; preparations of microbial, viral origin and their synthetic analogues: prodigiosan, zymosan, lycopid, etc.; preparations from human and animal tissues and their synthetic analogues: thymogen, splenin, pantocrine, blood and serum preparations, etc.; herbal preparations: tea tree, lemon, garlic, echinacea, etc.; synthetic immunomodulators: dibazole, methyluracil, levamisole, etc.; hormones: estradiol, synestrol, thyroxine, thyroidin, etc.; vitamins, microelements, and their salts) [3]. Many vitamins and minerals participate in immunological reactions, activating (or suppressing) the action of various enzymes, for example, selenium compounds, in particular sodium selenite [4], barium selenic acid [5], selenopyran [6].

The mechanisms of action of each immunostimulant are based on the features of its chemical structure, depending on which one or another system of the body is activated. At the same time, the pathogenesis of various diseases is not the same, so when choosing an immunostimulant it is necessary to have a clear idea of which systems are most involved in this pathology and which ones it can activate.

The combination of adverse organizational and technological factors with physiological changes increases the oxidative stress on the body of individual cows, which causes oxidative stress. In preliminary studies, we established the relationship between oxidative stress and immunobiochemical homeostasis in pregnant cows, indicating that the state of stress maladaptation lies in the etiopathogenesis of mass postpartum pathologies. In cows with obstetric pathology, a violation of immunobiochemical homeostasis was noted 15 days before calving (a decrease in hemoglobin, red blood cells, white blood cells, T and B lymphocytes, total blood serum protein (OBS), a violation of interfractional ratios, an increase in total lipids and cholesterol, a decrease the level of total immunoglobulins, vitamin E, lysozyme, bactericidal activity of blood serum and phagocytic activity of neutrophils) with a significant compared with cows with a normal course of labor and the postpartum period according to higher products of peroxidation (LP), in particular malondialdehyde. Therefore, the accumulation of highly active lipid peroxidation products in the body is one of the reasons leading to the suppression of factors of nonspecific immunological resistance of the body, i.e. immunological deficiency (table 1).

The technical result of the invention is the development of a method of immunocorrection in dry cows with the simultaneous prevention of violations of the reproductive function of cows in the postpartum period.

The technical result is achieved by the fact that in the method of immunocorrection, including the introduction of biologically active substances, antioxidants are used as biologically active substances, succinic and ascorbic acids are used, while succinic and ascorbic acids are introduced in the form of a mixture in a ratio of 1: 1, and the introduction is carried out per os within 20-25 days before calving and 20-25 days after calving. In addition, in this method, a mixture of antioxidants is administered at a dose of 1.0 g per head per day.

A comparison of the claimed technical solutions with the well-known ones made it possible to identify distinct features, therefore, these technical solutions meet the criterion of "novelty."

Table 1. Indicators of immunobiochemical homeostasis of cows 15 days before calving Indicators Norm Groups of cows with a normal course of the postpartum period with postpartum pathology Hemoglobin, g / l 99.0-129.0 101.0 ± 1.5 89.5 ± 1.8 Red blood cells, 10 ¹² / l 5.0-8.0 6.1 ± 0.2 4.7 ± 0.1 White blood cells, 10 ⁹ L 4,5-12,0 7.7 ± 0.4 5.0 ± 0.5 T-lymphocytes,% 45-60 42.9 ± 0.5 31.2 ± 1.9 B-lymphocytes,% 8-15 12.8 ± 0.8 7.4 ± 0.6 General immunoglobulins, g / l 15-25 18.9 ± 0.3 15.5 ± 0.2 Bactericidal activity of blood serum,% 65-85 66.8 ± 0.7 60.2 ± 1.1 Lysozyme activity of blood serum,% 6-9 6.9 ± 0.7 4.9 ± 0.5 Phagocytic activity of blood neutrophils,% 50-60 51.2 ± 1.5 45.2 ± 1.4 Total serum protein, g / l 72-86 76.0 ± 1.5 72.2 ± 1.7 Albumin,% 40 (30-50) 46.0 ± 1.5 41.2 ± 1.4 Globulins,% 60 - alpha 17 (12-20) 13.7 ± 0.9 12.1 ± 0.6 - beta 13 (10-16) 14.9 ± 1.1 13.2 ± 0.8 - gamma 30 (25-40) 25.4 ± 1.3 33.5 ± 1.5 Glucose, mmol / L 2.2-3.3 2.24 ± 0.2 2.09 ± 0.1 Total lipids, g / l 3.5-4.5 4.0 ± 0.4 5.9 ± 0.6 Total cholesterol, mmol / l 2,34-4,68 4.03 ± 0.02 12.9 ± 0.02 Vitamin E, μm / L 14-33 25.2 ± 1.2 12.9 ± 1.1 Malondialdehyde, μm / L 0.59-0.95 0.89-0.03 1.79 ± 0.02

Analysis of technical solutions showed that the claimed combination of essential features is unknown from the prior art, therefore, these technical solutions meet the criterion of "inventive step".

The feasibility and effectiveness of the method of immunocorrection was determined in scientific and industrial experiments on dry cows of black and white breed with a milk productivity of 3500-3800 kg in a farm with a high (over 90%) incidence of reproductive organs after calving. The state of immunobiochemical homestasis was assessed by the level of the following indicators: hemoglobin, erythrocytes, OBS, total immunoglobulins (determined by conventional methods), glucose (determined by the ortotoluidine method), total lipids (determined by Krinitsky in the modification of Volgin, 1969), lipid peroxidation products - diene conjugates and ketodenes (determined by Placert, 1976), malondialdehyde (determined with thiobarbituric acid), serum lysozyme activity (determined by V.G. Dorofeychuk), bactericidal activity of blood serum (op edelyali by O.V.Smirnovoy and T.A.Kuzminoy), phagocytic activity of blood neutrophils (determined by B.C.Gostevu).

The research results were subjected to biometric processing and correlation analysis using a computer program.

The essence of the method is illustrated by examples.

Example 1. The effectiveness of a mixture of succinic and ascorbic acids in a 1: 1 ratio as an immunocorrector was studied in 5 groups of dry cows formed on the basis of analogues, one of which was a control one. All animals were kept in the same conditions on diets balanced in nutrients, vitamins and minerals. For 20-25 days before the expected calving and 20-25 days after calving, the cows of the experimental groups were used inside with succinic acid and a mixture of succinic and ascorbic acids in a ratio of 1: 1 according to the following scheme: the first group - succinic acid in a dose of 1.0 g per head per day; the second, third and fourth groups: a mixture of succinic and ascorbic acids in a ratio of 1: 1 at a dose of 0.5, 1.0 and 1.5 g per head per day, respectively. Before the experiment and 2-3 days after calving, blood was taken to study immunobiochemical homeostasis. The results are shown in table 2.

Table 2. Immunocorrective efficacy of succinic and ascorbic acids (1: 1) INDICATORS Groups of cows 1st 2nd 3rd 4th 5th Succinic acid 1.0 g per head per day A mixture of succinic and ascorbic acids (1: 1), g per head per day Control (without drugs) 0.5 1,0 1,5 Hemoglobin, g / l 93.7 ± 2.0 94.4 ± 1.9 95.0 ± 2.5 93.9 ± 2.5 95.7 ± 3.0 112.0 ± 2.5 114.0 ± 1.1 119.0 ± 2.0 119.0 ± 2.0 98.0 ± 2.0 Red blood cells, 10 ¹² / l 5.4 ± 0.25 5.4 ± 0.1 5.2 ± 0.25 5.2 ± 0.25 5.2 ± 0.24 5.9 ± 0.3 6.1 ± 0.2 6.0 ± 0.2 6.0 ± 0.1 5.3 ± 0.15 OBS, g / l 72.0 + 1.3 72.3 + 1.5 70.7 ± 1.1 72.0 ± 1.4 72.2 ± 1.5 76.7 ± 1.4 77.3 ± 1.7 79.0 ± 1.2 79.0 ± 1.2 73.0 ± 1.1 Gamma globulins,% 23.1 ± 1.6 23.5 ± 1.2 23.0 ± 2.0 22.9 ± 1.5 23.9 ± 1.7 28.0 ± 1.3 28.5 ± 1.2 29.9 ± 1.3 29.8 ± 2.0 21.0 ± 1.4 Glucose, mol mol / l 2.03 ± 0.3 2.01 ± 0.2 2.09 ± 0.2 2.05 ± 0.2 2.15 ± 0.3 2.45 ± 0.2 2.46 ± 0.1 2.64 ± 0.2 2.58 ± 0.2 1.13 ± 0.4 Total lipids, g / l 5.69 ± 0.5 5.70 ± 0.5 5.61 + 0.3 5.65 ± 0.5 5.72 ± 0.5 4.98 ± 0.5 4.99 ± 0.4 4.87 ± 0.5 4.93 ± 0.4 5.96 ± 0.6 Lysozyme 3.3 ± 0.1 3.5 + 0.2 3.6 + 0.1 3.4 ± 0.3 3.5 ± 0.3 activity,% 4.2 ± 0.2 4.4 ± 0.2 4.8 ± 0.2 4.4 ± 0.2 2.0 ± 0.2 Bactericidal 64.4 ± 1.7 64.0 ± 1.6 63.0 ± 1.5 64.5 ± 1.8 65.5 ± 1.7 activity,% 73.0 ± 1.7 74.2 ± 1.7 79.7 ± 1.8 79.0 ± 1.9 60.7 ± 1.6 Phagocytic activity 45.7 ± 1.8 45.0 ± 1.4 43.5 ± 1.3 44.0 ± 1.5 45.8 ± 1.5 neutrophils,% 49.0 ± 0.2 51.0 ± 1.5 52.2 ± 1.5 51.5 ± 1.6 38.9 ± 1.4 Note: in the numerator - indicators for 20-25 days before calving; in the denominator - indicators for 2-3 days after calving

The obtained data confirmed the stimulating effect of succinic acid and a mixture of succinic and ascorbic acids in a ratio of 1: 1 on indicators of immunobiochemical homeostasis, which was most pronounced when using a mixture of succinic and ascorbic acids in a ratio of 1: 1, and it turned out to be dose-dependent. So, the average dose (1.0 g per head per day) provided the maximum indicators of immune activity. The maximum dose (1.5 g per head per day) provided indicators higher than the minimum (0.5 g per head per day), but lower or at the average dose level, which made increasing the dose inappropriate. The results obtained by testing the dose limits allowed us to recommend a dose of 1.0 g per head per day as optimal for practical use.

Example 2. The prophylactic effectiveness of a mixture of succinic and ascorbic acids in a ratio of 1: 1 in comparison with known methods was studied on 4 groups of dry cows formed on the basis of analogues, one of which was control. All animals were kept in the same conditions on diets balanced in nutrients, vitamins and minerals. In the 1st group, the cows were injected intramuscularly once with a 0.5% sodium selenite solution intramuscularly 20-25 days before calving (the analogous method), in the 2nd group, the cows were injected intramuscularly 20-25 days before and in the first 20 days after the calving tetravit once every 10 days (basic method), in the 3rd group, the cows were given a mixture of succinic and ascorbic acids in a ratio of 1: 1 at a dose of 1.0 g per day for 20-25 days before and 20-25 days after calving head per day. The animals were observed, noted the nature of the course of the prenatal, childbirth and the postpartum periods, performed clinical and gynecological examinations of cows, took into account the number of diseased animals, the timing of involution of reproductive organs and insemination. The state of immunobiochemical homeostasis was determined as in example 1. The results are presented in tables 3, 4.

Table 3. The state of immunobiochemical homeostasis Indicators Groups of cows 1st 2nd 3rd 4th Applied method Analog: sodium selenite intramuscularly Basic: intramuscular tetravitis Claimed: a mixture of succinic and ascorbic acids (1: 1) inside No drugs Hemoglobin, g / l 93.7 ± 2.0 94.0 ± 2.2 95.0 ± 2.5 95.7 ± 3.0 112.0 ± 2.5 101.0 ± 2.0 119.0 ± 2.0 98.0 ± 2.0 Red blood cells 5.4 ± 0.25 5.2 ± 0.3 5.2 ± 0.25 5.2 ± 0.24 10 ¹² / l 5.9 ± 0.3 5.7 ± 0.2 6.0 ± 0.2 5.3 ± 0.15 Total protein 72.0 ± 1.3 71.0 ± 1.2 70.7 ± 1.1 72.2 ± 1.5 serum, g / l 76.7 ± 1.4 75.2 ± 1.5 79.0 ± 1.2 73.0 ± 1.1 Gamma globulins, 23.1 ± 1.6 22.8 ± 1.3 23.0 ± 2.0 23.9 ± 1.7 % 28.0 ± 1.3 26.2 ± 1.5 29.9 ± 1.3 21.0 ± 1.4 Glucose, mmol / L 2.03 ± 0.3 2.20 ± 0.4 2.09 ± 0.2 2.15 ± 0.3 2.45 ± 0.2 2.87 ± 0.3 2.64 ± 0.2 1.13 ± 0.4 Lysozyme 3.3 ± 0.1 3.4 ± 0.3 3.6 ± 0.1 3.5 ± 0.3 activity,% 4.2 ± 0.2 4.0 ± 0.1 4.8 ± 0.2 2.0 ± 0.2 Bactericidal 64.4 ± 1.7 66.3 ± 1.6 63.0 ± 1.5 65.5 ± 1.7 activity,% 73.0 ± 1.7 75.5 ± 1.7 79.7 ± 1.8 60.7 ± 1.6 Phagocytic 45.7 ± 1.8 44.8 ± 1.4 43.5 ± 1.3 45.8 ± 1.5 neutrophil activity,% 49.0 ± 0.2 49.2 ± 1.5 52.2 ± 1.5 38.9 ± 1.4 Diene conjugates, 0.24 ± 0.02 0.24 ± 0.02 0.23 ± 0.03 0.22 ± 0.03 units A / ml 0.28 ± 0.02 0.32 ± 0.03 0.24 ± 0.01 0.36 ± 0.04 Ketodienes, units A / ml 0.055 ± 0.001 0.055 ± 0.001 0.056 ± 0.001 0.056 ± 0.001 0.065 ± 0.002 0.065 ± 0.002 0.061 ± 0.002 0.098 ± 0.001 Malonic 2.72 ± 0.03 2.73 ± 0.002 2.74 ± 0.02 2.72 ± 0.03 dialdehyde, µmol / l 2.76 ± 0.02 3.00 ± 0.03 2.76 ± 0.02 3.60 ± 0.03

Table 4. The comparative effectiveness of methods for the prevention of postpartum endometritis of cows Indicators Groups of cows 1st 2nd 3rd 4th Analog: sodium selenite Base: tetravit Declared: a mixture of succinic and ascorbic acids (1: 1) No drugs The number of cows subjected to preventive treatment, goal. 32 27 33 10 Sick postpartum endometritis, goal. eleven 16 3 10 Preventive efficacy,% 65.6 40.7 90.9 0 Dates of genital involution, days 26.7 ± 2.5 34.2 ± 4.5 23.3 ± 1.9 49.7 ± 5.0 The period from calving to insemination, days 64.4 ± 3.2 72.2 ± 3.7 48.2 ± 2.5 93.5 ± 8.5

The immunobiochemical status of cows 20-25 days before calving was characterized by low hemoglobin (3.3-5.6% below the lower limit of the physiological norm), erythrocytes (10-13% lower than the lower limit of the physiological norm) the level of the lower boundary of the physiological norm), gamma globulins (4.4-8.8% lower than the level of the lower boundary of the physiological norm), glucose (at the level of the lower boundary of the physiological norm), lysozyme, bactericidal activity of blood serum and phagocytic activity of blood neutrophils ( at the level of the lower gras physiological norm) against the background of a high level of lipid peroxidation products: diene conjugates (1.59-1.74 times higher than the physiological norm), ketodienes (1.08-1.1 times higher than the physiological norm), malondialdehyde (4, 59-4.64 times higher than the physiological norm). After calving, cows of the control group showed a further decrease in immunological reactivity (serum lysozyme activity decreased by 42.9%, serum bactericidal activity - by 7.3%, phagocytic activity of blood neutrophils - by 15%) with a more pronounced than experimental groups, increasing the performance of LPO products. So, the level of diene conjugates increased by 63.6% compared with 16.7% (analogue method), 33.3% (basic method) and 4.4% (the claimed method), the level of ketodienes - by 75.0% compared with 18.2% (analogue method), 54.2% (basic method), 8.9% (the claimed method), the level of malondialdehyde - 32.4% compared with 1.5% (method- analogue), 9.9% (basic method) and 0.7% (the claimed method). In all experimental groups, there was a tendency to normalize immunobiochemical parameters. More pronounced changes of a normalizing nature are established in the third experimental group (the claimed method). So, the lysozyme activity of blood serum increased by 33.3% compared with 21.9% (analogue method) and 17.7% (basic method), the bactericidal activity of blood serum increased by 26.5% compared to 14.8 % (analogue method) and 13.9% (basic method) and phagocytic activity of blood neutrophils - by 20% compared with 8% (analogue method) and 9.8% (basic method) (table 3). The effectiveness of the prevention of postpartum endometritis in cows in the third group was 90.9% compared with 65.6% in the first group (analogue method), 40.7% in the second group (basic method) and 0% in the fourth group (table 4 )

The results of the studies confirmed that when implementing the claimed combination of essential features, a technical result is achieved - immunocorrection in dry cows, manifested by the normalization of the main indicators of immunobiochemical homeostasis, including indicators of natural resistance, which ensures the normal course of labor and the postpartum period.

Information sources

1. Pritulin P.I., Kalmykova T.P. Infectious problems of animal immunodeficiency // Bulletin of agricultural science. - 1989. - No. 2. - S.95-100.

2. Ignatov P.E. Immunity and infection. Management capabilities. - M .: Time, 2002 .-- 350 s.

3. Theory and practice of the use of biologically active substances in animal husbandry, Kirov, October 6-7, 1998: Abstracts. doc. scientific conf. Kirov, 1998 .-- 98 s.

4. Sokolov V.D. Pharmacology, 2nd ed., Revised and supplemented. M .: Kolos, 2000 .-- S.360-361.

5. RU 2201237 C2, 03/27/2003.

6. RU 2212888 C2, 09/07/2003.

Claims (2)

1. The method of immunocorrection in cows, including the introduction of a mixture of succinic and ascorbic acids in a ratio of 1: 1 in a dose of 0.5 to 1.5 g per head per day per os for 20-25 days before and 20-25 days after calving . 2. The method according to claim 1, characterized in that succinic and ascorbic acids are administered in a dose of 1.0 g per head per day.