Method for obtaining medication for stimulating non-specific resistance and realizing reproductive qualities of cows

Abstract

FIELD: biotechnology; veterinary medicine.SUBSTANCE: method for producing a drug for stimulating non-specific resistance of the body and reproductive qualities of cows. The method is characterized by thoroughly mixing 10 ml of an oil solution of vitamin E containing 2.5 g of α-tocopherol acetate, with 0.15 g of emulsifier polysorbate 80 to obtain an oil solution, then 3.0 ml of purified concentrate of yeast cell polysaccharide complex is added to 44 ml of phosphate buffer solution and constantly stirred until completely dissolved to obtain a buffer solution of purified concentrate of yeast cell polysaccharide complex. A buffer solution of selenomethionine is then obtained by dissolving 125 mg of the organic form of selenium in the form of selenomethionine in 44 ml of phosphate buffer solution. The resulting buffer solutions are combined and mixed with the oil solution. The resulting emulsion is filtered through a sterilization filter with a pore size of 0.45 µm and poured into a 100 ml bottle.EFFECT: use of the invention will optimize metabolic processes and improve the reproductive performance of cows.1 cl, 3 tbl, 3 ex

Description

[0001] The invention relates to the field of biotechnology and veterinary medicine, namely to methods for producing drugs for stimulating non-specific resistance and reproductive qualities of cows.

[0002] A method for obtaining a biologically active preparation from yeast (SU 1808331 A1 A61K 35 / 72, 15.04.93) in the form of a purified polysaccharide complex with immunostimulating activity is known. Immobilization of yeast cell polysaccharides in agar gel (TU 10.07.236-91 for the production of the drug "Dostim") or in a 10-14% aqueous-salt solution of polyvinylpyrrolidone (RU 2137480 C1 A61K 31 / 79, 35 / 72, 20.09.99) increases the immunostimulating activity of the drugs.

[0003] The closest analogue (prototype) for the problem being solved is a method for obtaining a drug for increasing non-specific resistance and immunogenesis of the organism of farm animals and poultry, including mixing 0.2-0.3% agar suspension and a concentrate of purified polysaccharide complex of yeast cells. (-)2,3,5,6-tetrahydro-6-phenylimidazo-[2,1-b]-thiazole hydrochloride and formalin are added to the resulting mixture with constant stirring, the volume is brought to 100 ml with distilled water. The drug has immunostimulating activity (RU 2332214 C1 A61K 31 / 429, 31 / 115, A61K 36 / 06, A61K 36 / 02, A61P 37 / 04, 27.08.2008).

[0004] The vitamin and mineral complex E-selenium is widely used in practice. It is also aimed at preventing non-infectious immunodeficiencies caused by stress, infections, invasions, vaccination, deworming, and poisoning. 1 ml of the preparation contains the following active ingredients: selenium (as sodium selenite) - 0.5 mg and vitamin E - 50 mg, as well as excipients: polyethylene-35-ricinol, benzyl alcohol, and water for injection - up to 1 ml (TU 9353-016-34214729-01 with amendments No. 1, No. 2; PVR-3-4.1 / 00814).

[0005] The disadvantages of PS-2 (RU 2332214) include its monodirectional action: the yeast cell polysaccharide complex has limited immunostimulatory action, lacks vitamin E and selenium, and has poor bioavailability due to the excipient (agar). Disadvantages of the E-selenium complex include a less bioavailable form of selenium, sodium selenite, and the lack of an immunostimulatory component.

[0006] The invention aims to stimulate non-specific resistance and reproductive performance in cows. Stimulation of non-specific resistance, achieved through the synergistic effect of a polysaccharide complex, α-tocopherol acetate, and selenomethionine, promotes the activation of cellular and humoral immune factors, ensuring the effective prevention of diseases caused by secondary immunodeficiencies. The expansion of the arsenal of immunomodulatory agents is achieved through the use of natural components with proven biological activity. Increasing the bioavailability of lipophilic vitamin E through emulsification and the use of an organic form of selenium (selenomethionine) optimizes their absorption and utilization in the body. Together, these factors lead to the optimization of metabolic processes and improved reproductive performance. The technical result is as follows:

[0007] 1) Thoroughly mix 10 ml of vitamin E oil solution (2.5 g of α-tocopherol acetate) with 0.15 g of polysorbate 80 emulsifier;

[0008] 2) Add 44 ml of phosphate buffer solution to 2.5-3.0 ml of purified yeast cell polysaccharide complex concentrate, stirring constantly until completely dissolved;

[0009] 3) 125 mg of organic form of selenium as selenomethionine L-Selenomethionine-(methyl- 13 C) dissolve in 44 ml of phosphate buffer solution;

[0010] 4) combine the buffer solutions of purified yeast cell polysaccharide complex concentrate and selenomethionine;

[0011] 5) mix the oil solution of vitamin E and the emulsifier polysorbate 80 with a buffer solution of polysaccharides and selenomethionine;

[0012] 6) filter the finished emulsion through a sterilization filter with a pore size of 0.45 µm;

[0013] 7) pour the resulting preparation into a 100 ml bottle;

[0014] 8) After sterilization, use to stimulate non-specific resistance and reproductive qualities of cows.

[0015] Unlike known analogues, the proposed solution has the advantage of maintaining its effect on non-specific resistance factors while simultaneously administering fat-soluble vitamin E and an organic form of selenium (selenomethionine) along with a yeast cell polysaccharide complex. This is achieved because vitamin E, by regulating metabolic processes and potentiating the effects of vitamins A and D3, supports immunity, while selenium, with its antioxidant properties, promotes detoxification and enhances the immune status of animals, providing a synergistic effect.

[0016] The following substances were used to implement the method:

[0017] 1) oily α-tocopherol acetate Reg. 77-3-29.13-1666 No. PVR-3-0.2 / 01126 dated 01.11.23;

[0018] 2) Purified yeast polysaccharide complex concentrate SU 1808331;

[0019] 3) selenomethionine - CAS 1217470-45-5;

[0020] 5) emulsifier polysorbate 80 - CAS 9005-65-6;

[0021] 6) phosphate buffer solution - GOST 4919.2-2016.

[0022] The method is as follows.

[0023] Example 1. Thoroughly mix 10 ml of vitamin E oil solution (2.5 g of α-tocopherol acetate) with 0.15 g of polysorbate 80 emulsifier; add 44 ml of phosphate buffer solution to 2.5 ml of purified yeast cell polysaccharide complex concentrate, stirring constantly until completely dissolved; dissolve 125 mg of organic selenium in the form of selenomethionine in 44 ml of phosphate buffer solution; combine the buffer solutions of purified yeast cell polysaccharide complex concentrate and selenomethionine; mix the vitamin E oil solution and polysorbate 80 emulsifier with the buffer solution of polysaccharides and selenomethionine; filter the finished emulsion through a sterilization filter with a pore size of 0.45 μm; pour the resulting preparation into a 100 ml bottle; After sterilization, use to stimulate non-specific resistance and reproductive qualities of cows.

[0024] Example 2. Thoroughly mix 10 ml of vitamin E oil solution (2.5 g of α-tocopherol acetate) with 0.15 g of polysorbate 80 emulsifier; add 44 ml of phosphate buffer solution to 3.0 ml of purified yeast cell polysaccharide complex concentrate, stirring constantly until completely dissolved; dissolve 125 mg of organic selenium in the form of selenomethionine in 44 ml of phosphate buffer solution; combine the buffer solutions of purified yeast cell polysaccharide complex concentrate and selenomethionine; mix the vitamin E oil solution and polysorbate 80 emulsifier with the buffer solution of polysaccharides and selenomethionine; filter the finished emulsion through a sterilization filter with a pore size of 0.45 μm; pour the resulting preparation into a 100 ml bottle; After sterilization, use to stimulate non-specific resistance and reproductive qualities of cows.

[0025] Example 3. The subjects of the study were cows in the dry period (60 days before the expected calving), postpartum, and lactation periods. For the scientific and farm experiment, four groups of cows (control, 1st, 2nd, 3rd, and 4th experimental) were selected based on the analog principle, taking into account the clinical and physiological condition, age, and live weight of 10 animals in each group. The animals were included in the experiment from the moment of weaning until the end of lactation.

[0026] The cows of the 1st experimental group were injected intramuscularly with the PS-2 prototype at a dose of 10.0 ml per head three times 60 and 30 days before the expected calving and on the day of calving; the cows of the 2nd experimental group were injected with the E-selenium prototype at 10.0 ml intramuscularly 60 days before calving and on the day of calving; the 3rd experimental group was given the preparation according to the invention (example 1); the 4th experimental group was given the preparation according to the invention (example 2) at a dose of 10.0 ml per head three times 60 and 30 days before the expected calving and on the day of calving (experimental variants); the fifth group was a control group.

[0027] Indicators of nonspecific resistance in cows were determined 30-25 days before and 5-10 days after calving (Table 1). In the post-calving period, the incidence of reproductive organ diseases and metabolic disorders were studied (Table 2).

[0028]

[0029] It was established that the prototype (PS-2) and prototype (E-selenium), as well as the proposed preparations according to examples 1 and 2, had a positive effect on the indicators of non-specific resistance of the cows’ organism, as evidenced by the data presented in the table.

[0030] 30-25 days before calving, the following changes were observed: phagocytic activity in the 1st experimental group (PS-2) was 2.86% higher than the control, while in the 2nd experimental group (E-selenium) it was slightly lower. In the groups that received the drugs according to Examples 1 (p <0.05) and 2 (p <0.01), a significant increase in this indicator was noted - by 6.32% and 8.45%, respectively. Lysozyme activity of blood serum also demonstrated positive dynamics: in the 1st experimental group (PS-2) it was higher by 5.76%, in the 2nd (E-selenium) - by 7.19%, and in the groups that received the drugs according to Examples 1 and 2 - by 7.91% and 10.79%, respectively. The bactericidal activity of blood serum showed a similar trend: an increase of 2.95% in the 1st experimental group (PS-2), by 1.26% in the 2nd (E-selenium), and a more pronounced increase in the groups that received the drugs according to examples 1 (p<0.05) and 2 (p<0.01) - by 3.79% and 5.26%, respectively.

[0031] In the period 5-10 days after calving, the pattern of changes remained similar. Phagocytic activity in the 1st experimental group (PS-2) was higher than in the control group by 4.95%, in the 2nd (E-selenium) - slightly lower. The preparations according to examples 1 (p <0.01) and 2 (p <0.001) demonstrated a significant increase in this indicator - by 11.81% and 14.19%, respectively. Lysozyme activity in the 1st experimental group (PS-2) increased by 5.56%), in the 2nd (E-selenium) - by 3.97%, and in the groups receiving the preparations according to examples 1 (p <0.01) and 2 (p <0.001) - by 13.49% and 15.87%, respectively. The bactericidal activity of blood serum showed an increase of 4.13% in the 1st experimental group (PS-2), by 1.52% in the 2nd (E-selenium), and a more pronounced increase in the groups that received the drugs according to examples 1 (p<0.05) and 2 (p<0.01) - by 5.22% and 6.30%, respectively.

[0032]

[0033] An analysis of cow disease incidence in the experimental and control groups revealed that the use of the study drugs in the experimental groups resulted in a reduction in disease incidence compared to the control group. The drug in Example 2 stands out, demonstrating a significant reduction in disease incidence across a number of indicators. For example, the drug in Example 2 completely eliminated cases of clinical mastitis and aseptic claw disease, reducing the incidence by half compared to the control group. Furthermore, this group saw a 75% reduction in the incidence of ketosis, from 4 cases in the control group to 1 case in the experimental group, and a 75% reduction in the incidence of uterine subinvolution, from 4 cases in the control group to 1 case in the experimental group. The drug in Example 1 also demonstrated high efficacy in reducing clinical mastitis and abomasal displacement, completely eliminating these conditions.

[0034]

[0035] The results of the study revealed that in all experimental groups, with the exception of the first (prototype PS-2), there is a tendency towards an earlier manifestation of estrous activity, characterized by a decrease in the period before the onset of the first estrus, compared to the control group (38.1 ± 2.15 days). The most significant reduction in this indicator was recorded in the 4th experimental group (31 ± 2.10 days), which is confirmed by statistically significant differences relative to the control group (p < 0.05), which is associated with a low incidence of cows in the postpartum period compared to other groups of animals. The maximum value of the fertilization index was recorded in the control group - 2.6, which is higher than in the 3rd and 4th experimental groups, by 1.2 and 1.3 times, respectively. A reduction in the service period was noted in all experimental groups compared to the control group (111.5±5.33 days), with the minimum value established in the 4th experimental group (100.1±3.84 days).Higher conception rates were observed in experimental groups 3 and 4 (86.6%) compared to the control group (73.3%). Analysis of the number of inseminations required to achieve pregnancy indicates that most cows are fertilized within the first three estrous cycles, with a higher number of successfully fertilized cows in experimental groups 3 and 4 (13 heads) compared to the control group (11 heads).

[0036] Thus, the results of the study indicate a positive effect of the studied drugs, developed on the basis of examples 1 and 2, on the indicators of non-specific resistance and reproductive parameters of cows.

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