RU2377999C2 - Method for production of biostimulant from cow placenta - Google Patents

Abstract

FIELD: veterinary science.

SUBSTANCE: invention relates to veterinary medicine and is to be utilised for prophylactics and treatment of cow reproductive malfunctions. The method envisages cow placenta preparation. The prepared cow placenta is chilled. Then the placenta having been chilled is homogenised and exposed to microwave radiation at a frequency of 2450±50 MHz, with the raw material heated to a temperature of no more than +60°C. Then the resultant liquid is mixed with distilled water at a ratio of 1:4 and boiled during 10 minutes. Then the resultant liquid is chilled, centrifuged and filtered. Then the resultant liquid is mixed with Fraction 2 Dorogov antiseptic stimulant, bringing the concentration of Fraction 2 Dorogov antiseptic stimulant to 4% of the total liquid volume; after that one adds a 20% alcoholic solution of thymol in an amount of 0.1% of the total liquid volume. Then the resultant preparation is autoclaved.

EFFECT: proposed invention enables improvement of cow impregnation capacity after initial insemination.

1 tbl, 2 ex

Description

The invention relates to veterinary medicine, and in particular to a technology for producing preparations used for the prevention and treatment of impaired reproductive function of cows.

A known method of obtaining a biostimulator "POM" from the placenta of cows. The placenta is cooled, homogenized and extracted with methyl urea at a urea to formaldehyde ratio of 4: 2, taking 40-50% of the placenta mince from the weight of urea. The components are mixed, heated to 100 ° C, adjusted to pH 8.0-10.0, cooled to 60-70 ° C, distilled water is added to 1 L and sedimented. The method prevents retention of the placenta by 66.4% and the incidence of endometritis by 48.6% more than in the control (SU 1730762 A1, A61K 35/12, 07.20.1996). However, the preparation obtained using this technology contains a significant amount of products of nitrogen metabolism (urea), which have a toxic effect on the tissues of the animal both at the injection site and on the whole organism.

A known method of obtaining anti-inflammatory drugs from the placenta of the first half of pregnancy of cows. The placenta is crushed, homogenized, subjected to selective extraction, then the extract is disinfected, autolysis, aseptic filtration. The result is a drug that does not have side effects and allergic manifestations during its therapeutic use (RU 2128514 C1, A61K 35/50, 1999). However, the preparation of this drug is associated with the use of the placenta of early pregnancy, which in practice is often not feasible.

A known method of producing a biostimulator from the placenta of cows, according to which the liquid component from the placenta is separated by heating in a microwave oven at a radiation frequency of 2450 ± 50 MHz in a special dish with a perforated bottom, while the liquid component is separated by gravity, and the heating is performed no higher than 60 ° C. moreover, as a dilution medium for the liquid fraction using a 0.9% solution of sodium chloride. The method allows to reduce the number of technological operations and the total time to obtain the drug while maintaining its quality (RU 2138273 C1, A61K 35/50, 1999).

The closest in technical essence and the achieved result is the technology for the preparation of Biostimulgin - microwave, designed to prevent phytoplacental insufficiency in cows (Kolchina AF Diseases of pregnant women and perinatal pathology in animals / Monograph. - Yekaterinburg, Ural State Agricultural Agricultural Academy. - Yekaterinburg, 1999. - S.144).

The last cows of 2-4 calving were kept for four days at 2-4 ° C, after which they were subjected to microwave irradiation with a frequency of 2450 ± 50 MHz in a microwave oven when the raw material was heated no higher than 60 ° C. As a result, 30 to 60% of the liquid fraction of the mass of the afterbirth was obtained. Warming up was done in cycles of 15-20 minutes. The combined fractions were diluted with 0.9% sodium chloride in a ratio of 1: 4 and mixed thoroughly. The pH of the resulting solution was adjusted to 7.75-8.40 and boiled for 10 minutes. The cooled suspension was centrifuged, filtered, packaged in 100-200 ml vials, sterilized by autoclaving at 1.5 atm for one hour.

In cows treated with Biostimulgin - microwave subcutaneously at a dose of 20 ml twice a day 10 days before calving and in the first hours after birth, the fertility from the first insemination in the experimental group was 38.2%, in the control group - 52.9%. The insemination index in the cows of the experimental group was 1.58 and was lower than in the control by 0.18, that is, it was preferable, the service period was shorter by 15.42 days. However, according to the prototype, the fertility of cows from the first insemination in the experimental group was lower than in the control by 14%. With a first insemination index of 1.58, it follows that the success rate of subsequent inseminations, amounting to 58.82%, was more effective than immediately after using the drug. Practice shows that the first insemination is more effective than subsequent ones. The method is not effective enough when applied to highly productive animals. In addition, inflammatory processes occur in cows at the injection site.

The technical result of the claimed invention is to increase the effectiveness of the first insemination after calving. The technical result of the invention is achieved by the fact that a method for producing a biostimulator from the placenta of cows is provided, which provides for cooling the placenta of cows, treating the placenta in a microwave oven at a radiation frequency of 2450 ± 50 MHz, heating the feed at a temperature of not more than 60 ° C, mixing the liquid released from the feed (product ) with a diluent in a ratio of 1: 4, boiling for 10 minutes, cooling, subsequent centrifugation of the obtained liquid, its filtration and autoclaving, characterized in that before processing the afterbirth it is homogenized in a microwave oven, distilled water is used as a diluent, after filtration, the product is mixed with ASD fraction 2, bringing the concentration of ASD fraction 2 to 4% of the total liquid volume, add a 20% alcohol solution of thymol in an amount of 0.1% of total volume of fluid.

The resulting preparation is called "Placentin-A".

Example 1. "Placentin-A" was prepared as follows:

the raw placenta separated for 4-6 hours after calving was used as a raw material for the preparation of the preparation without visible pathological changes in tissues, taken from healthy cows of 2-4 calving;

the raw materials were placed in the refrigerator for 48-72 hours at 4-5 ° C;

after that, the feed was homogenized and subjected to microwave exposure with a radiation frequency of 2450 ± 50 MHz when the feed was heated not higher than 60 ° C to separate the liquid component;

the resulting liquid was mixed with distilled water in

proportions 1: 4 and boiled for 10 minutes to denature the proteins;

after cooling, the resulting suspension was centrifuged, filtered, mixed with ASD fraction 2, bringing the concentration of ASD fraction 2 to 4% of the total liquid volume;

to the resulting mixture was added a 20% alcohol solution of thymol in an amount of 0.1% of the total liquid volume. The introduction of thymol provides an increase in the duration of storage of the drug in connection with its bacteriostatic effect and prevents the occurrence of an inflammatory process at the injection site;

the resulting preparation was poured into vials and autoclaved at a pressure of 2 atmospheres for 15 minutes.

The action of the drug is based on the complex effect of low molecular weight peptides and free nucleotides that can stimulate metabolic processes and the general resistance of the body. In addition, the placenta contains a number of biologically active substances, including progesterone, estradiol and its derivatives - estrol and estriol, which actively affect the proliferative processes in the endometrium and the tone of the smooth muscles of the uterus.

The preparation ASD of fraction 2, which is a part of Placentin-A, is a product of dry distillation of raw materials of animal origin. It contains compounds with an active sulfhydryl group, derivatives of aliphatic amines, carboxylic acids, aliphatic and cyclic carbohydrates, derivatives of amides and water. ASD fraction 2 is a powerful immunomodulator that allows you to normalize the immune response of the mother. In addition, this drug has an antiseptic effect, which increases the shelf life of Placentin-A.

Example 2. In the GPZ them. Frunze of the Belgorod region at the Bessonovsky dairy complex studied the effectiveness of using Placentin-A to increase the reproductive function of cows of black-motley breed of various productivity in the postpartum period. For this purpose, four experimental groups of 20 goals each were created by the principle of analogues. Group No. 1 was formed from cows with a productivity of 3500-4000 kg of milk per lactation, group No. 2 - 4000-4500 kg, group No. 3 - 4500-5000 kg, group No. 4 - 5000-5500 kg. Cows of all experimental groups 20-25 days before calving and immediately after calving were once administered subcutaneously "Placentin-A" in a dose of 20 ml. The use of "Placentin-A" 20-25 days before calving is due to the fact that this allows you to enhance chemotaxis in placentomas and thereby create conditions for faster maturation and subsequent excretion of the placenta. And the estrogens contained in the extract enhance the contractility of the smooth muscles of the uterus. The use of "Placentin-A" immediately after calving is also due to the specific action of estrogens, which reduces the time of uterine involution and prevents the development of postpartum endometritis. For each experimental group, a control was selected from animals with identical productivity, not subjected to processing, similar in age and conditions of detention. The condition of the uterus and ovaries was determined by rectal examination. In addition to the terms of uterine involution, indicators such as the time of the first hunting after calving, the level of pregnancy after the first insemination, the length of the service period and the insemination index were taken into account. The results are presented in the table.

Table
The effect of "Placentin-A" on the timing of uterine involution and restoration of sexual cyclicity in cows at various levels of productivity Groups Number of animals
in Group The average timing of the first involution of the uterus, days The average timing of the first hunt, days Pregnant after the first insemination Service period, days Index
insemination goals %, M ± m Group No. 1 - 3500-4000 kg twenty 22.4 ± 0.90 * 22.3 ± 1.89 fifteen 75 ± 11.1 84.5 ± 1.98 2.1 Control group twenty 28.3 ± 0.87 26.4 ± 2.76 12 60 ± 14.1 81.0 ± 2.43 2.7 Group No. 2 - 4000-4500 kg twenty 25.5 ± 1.11 * 27.2 ± 1.94 12 60 ± 14.1 75.6 ± 1.87 * 1.9 Control group twenty 38.8 ± 0.76 30.0 ± 1.90 eleven 55 ± 15.0 87.9 ± 2.20 2.5 Group No. 3 - 4500-5000 kg twenty 31.2 ± 0.92 * 27.5 ± 1.00 ** fourteen 70 ± 12.2 88.5 ± 2.82 1.9 Control group twenty 46.7 ± 1.43 31.5 ± 1.87 9 45 ± 16.5 93.3 ± 2.51 2.9 Group No. 4 - 5000-5500 kg twenty 33.6 ± 0.85 * 33.7 ± 2.81 13 65 ± 13.2 89.0 ± 1.95 * 2.2 Control group twenty 51.3 ± 1.37 38.5 ± 2.97 9 45 ± 16.5 115.2 ± 2.54 3,1 * P <0.001; ** P <0.05

As can be seen from the data in the table, in all experimental groups, the periods of uterine involution were lower than in the control (P <0.001). The first hunt in the experimental groups occurred 2.8-4.8 days earlier than in the control. The pregnancy level after the introduction of "Placentin-A" was higher than in the control groups by 5-25%. The average service period in the experimental groups was 12.3-26.2 days shorter than in the control. The difference between the duration of the service period of the experimental groups No. 2, No. 4 and the control was significant (P <0.001). The exception is group No. 1, where the service period was 3.5 days longer than in the control, but the difference is not significant. The insemination index in the experimental groups was also lower than the control. It should be noted that with an increase in productivity in both experimental and control groups, the periods of uterine involution, the time of manifestation of the first hunting after calving, the service period and the insemination index increased. At the same time, there was a decrease in the level of pregnant after the first insemination of animals. But in all cases, the indicators of the experimental groups were better than the control.

At the injection site of the drug, no inflammatory processes were detected. Compared with the prototype, the use of "Placentin-A" provides higher performance of the first insemination, which, depending on the productivity of the cows, ranged from 55 to 75% in the experimental groups, which is 5-25% more than in the control. In the prototype, the fertility of cows after the first insemination in the experimental group was 38.2%, in the control - 52.9%.

The invention is applicable to enterprises for the breeding of herds of dairy and dairy-meat cows.

Claims (1)

A method of producing a biostimulator from the placenta of cows, which involves cooling the afterbirth of cows, treating the afterbirth in a microwave oven at a radiation frequency of 2450 ± 50 MHz, heating the feed at a temperature of no more than + 60 ° C, mixing the liquid (product) released from the feed with a diluent in the ratio of 1: 4, boiling for 10 min, cooling, subsequent centrifugation of the obtained liquid, its filtration and autoclaving, characterized in that before processing the subsequent cows in a microwave oven, it is homogenized as a diluent and distilled water is used, after filtration the product is mixed with the ASD of fraction 2, the concentration of the ASD of fraction 2 is adjusted to 4% of the total liquid volume and a 20% alcohol solution of thymol in the amount of 0.1% of the total liquid volume is added.