RU2431491C1 - Method for producing preparation of boar natural sex pheromones - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to veterinary science, particularly to production of a preparation of boar natural sex pheromones. A method for producing the preparation of boar natural sex pheromones, involving mixing pericarps, root of celeriac, root of parsnip in certain proportions, grinding, homogenising, adding the enzyme protosubtiline, incubating, centrifuging, mixing with urine of senior boars in certain proportions, pressing out and sublimating the produced fluid.

EFFECT: method allows providing higher yield of the preparation with better biological activity.

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Description

The invention relates to the field of veterinary medicine, in particular to the production of biological preparations that are used in pig farming as sexual function stimulants in sows.

A known method of producing a preparation of sex pheromones from bull urine (Auth. St. No. 1666102, 1991 "A method of producing sex pheromones", auth. Kononov VP and others) and a method of producing a preparation of natural sex pheromones of a boar (Patent No. 2034521, 1992 “A method for producing a preparation of natural sex boar pheromones”, author Sein O. B. et al.). However, these methods, complex in execution, require special equipment and qualified support. At the same time, the resulting preparations of sex pheromones have low biological activity.

The closest technical solution, selected as a prototype, is a method for producing a preparation of natural sex pheromones of a boar (Patent No. 2159597, 1999 “A method for producing a preparation of natural sex pheromones of males”, author Sein OB and others), which consists in mechanical grinding of testes obtained from sexually mature males to a minced state, followed by homogenization to a homogeneous mass. In order to increase the destruction of cell tissue and the release of pheromones into the solution, the protosubtilin enzyme (GZH GOST 23636-79) is added to the homogenizate in an amount of 1-1.5% of the total weight of the feedstock and the resulting mixture is incubated at a temperature of 37-39 ° С for 50-60 minutes, and then centrifuged until separation into two fractions. The supernatant is mixed with male urine in a ratio of 1: 2-2.5 and distilled with water vapor. The condensate obtained is a preparation of natural male sex pheromones. Shelf life 12 months.

The disadvantage of this method is the low yield of the final product, relatively low biological activity, short storage period.

The aim of the invention is to increase the yield of the drug with high biological activity and a long shelf life.

This goal is achieved by the fact that the testes and urine of sexually mature boars, as well as root celery root and parsnip root, containing steroids 5α-androst-16-en-3-one and 5-α-androst-16-en 3α-ol, which are the basis of sexual boar pheromones. The seeds and roots of celery and parsnip in a ratio of 1.0: 0.25: 0.25 are ground to a minced state and homogenized in a homogenizer to a homogeneous mass. Then, the protosubtilin enzyme (GZH GOST 23636-79) is added to the homogenizate in an amount of 1.0-1.5% of the total weight of the feedstock. The resulting mixture is incubated in a thermostat at a temperature of 37-39 ° C for 50-60 minutes, after which it is mixed with urine of mature boars in a ratio of 1: 1.0-1.5 and squeezed using a press with 0.7 × 0 cells. 7 mm. The resulting liquid is poured into glass vials of 15.0 ml and subjected to sublimation. After sublimation, the vials are closed with rubber stoppers and rolled in aluminum caps.

The optimal ratio of these components was selected using mathematical design of the experiment using the steep climb method. With an increase in the amount of urine, a decrease in the concentration of the drug by 10-15% occurs, and with a decrease in the urine of the drug, they receive 25-30% less than with the optimal ratio of components.

The optimal incubation regimen was selected empirically. At temperatures below 37 ° C and above 39 ° C, the activity of the protosubtilin enzyme decreases, as a result of which the hydrolysis process is not active enough, which leads to a decrease in the yield of the drug. When the incubation time is less than 50 min, the yield of the final product decreases by 10-15%, which is associated with incomplete hydrolysis. With an increase in incubation time of more than 60 min, the saturation of the drug with pheromones does not increase, since the hydrolysis process is completed by this time and the pheromone enters the solution.

After carrying out all stages of the technological process, the preparation is a dry porous mass, brownish in color with a specific smell. Shelf life 24 months at a temperature not exceeding 20 ° C.

Before using the drug, the vial is freed from the aluminum cap and cork, the vial is diluted in 100 ml of water with a temperature of 35-40 ° C, filtered through 2-3 layers of a gauze filter and sprayed with spray at the sows at the level of the nasal mirror of animals.

Example. To 1000 g of testes obtained from mature boars and purified of connective and adipose tissue, 250 g of peeled root celery root and 250 g of seed parsnip were added, ground to a minced state and homogenized in a homogenizer to a homogeneous mass. 15 g of protosubtilin enzyme were added to the resulting mass and incubated in a thermostat at a temperature of 37-39 ° С for 60 min. Then 1515 ml of boar urine was added to the homogenizer, thoroughly mixed and squeezed using a press with 0.7 × 0.7 mm cells. The resulting liquid was poured into 15 ml glass vials and sublimated. After the sublimation process, the preparation was a dry porous mass (0.5 g) of a brownish tint with a specific odor. Vials with the drug were closed with rubber stoppers and rolled in aluminum caps. The drug was stored at 20 ° C, a shelf life of 24 months.

Before using the drug, it was dissolved in 100 ml of water with a temperature of 35-40 ° and filtered through a gauze filter.

The drug was tested for sterility using MPA and Endo media, followed by incubation in a thermostat. The toxicity of the drug was checked on 10 repair mumps of large white breed of 5 months of age, which were placed in a special chamber-machine and sprayed using an aerosol generator. During and after treatment of repair pigs with a preparation of natural sex pheromones of the boar, behavioral reactions, their main clinical and hematological parameters were within the physiological norms. Guinea pigs (10 animals) after the intraperitoneal administration of the drug (0.5 ml / animal) did not show toxicosis, all experimental animals were alive.

The biological activity of the prepared preparation was determined using a biological test and using a special olfactometer. The manufactured preparation was compared with the prototype preparation (taking into account the shelf life), urine of a mature boar, extract from the root of the celery root, extract from the root of the sowing parsnip, decoction of compound feed, distilled water. During the biotest, an ethogram was filled out, which included behavioral responses in sows during contact with the test object. The results of the biotest showed (Table 1) that in sows, the percentage of key responses to the manufactured drug was higher (89.5%) than to the prototype drug (78.8%).

Production tests of the manufactured preparation were carried out under the conditions of the Znamenskoye experimental-experimental farm of the Kursk State Agricultural Academy named after I.I. Ivanov, and the pig breeding complexes Magnitny and Timsky of the Kursk Region.

The first scientific and production experiment was carried out on immature repair pigs, analogues of large white breed of 5 months of age. Three groups of 35 goals were formed. Group 1 pigs were stimulated with an experimental preparation. Group 2 pigs were treated with the prototype drug. Group 3 pigs were control, they were treated with distilled water.

Used preparations of sex pheromones in a dose of 0.5 ml per animal, by spraying with a spray at the level of the nasal mirror, every other day for 20 days. Experimental and control animals were kept in the same room and received the same diet. Guinea pig sex was determined using a probe boar.

The results of the first scientific and production experiment showed that puberty in pigs of the 1st experimental group occurred at 181.5 ± 2.03 days old, in pigs of the 2nd experimental group - from 187.0 ± 1.85 days, and in pigs 3 the control group - from 205.5 ± 2.11 days. In most guinea pigs subjected to pheromone stimulation, puberty occurred in the first 10 days after the start of stimulation, and in guinea pigs puberty was delayed and only 16 (46%) animals showed a 30-day observation period. Moreover, in pigs, which were stimulated with the manufactured preparation, the number of sexually mature animals was 31 (89%) compared with animals stimulated by the prototype preparation - 28 (80%) (Table 2).

The second scientific and production experiment was carried out on repair mumps, analogues of 8 months of age, which had a good development, but they did not puberty. From these pigs, three groups of 15 animals were selected. Pigs of the 1st experimental group were treated with the prepared preparation of natural sex boar pheromones. Pigs of the 2nd experimental group were stimulated with the prototype drug. Group 3 mumps were control and were not subjected to pheromone stimulation. Pheromone stimulation was carried out daily for 20 days, the dose of the drug was 0.5 ml / goal. Experimental animals were observed for 30 days, sexual hunting was determined using a probe boar.

The results of the second scientific and production experiment showed that after stimulation with the manufactured drug, puberty occurred in 11 (73%) pigs. When using the prototype drug, maturity appeared in 9 (67%) pigs, and in the control group in 3 (20%) pigs (Table 3.).

The third scientific and production experiment was carried out on lactating sows of large white breed. Three groups of 30 sows each were formed. Sows 1 of the experimental group were subjected to stimulation with the manufactured drug. Sows 2 of the experimental group were stimulated with the prototype drug. Sows of group 3 were the control and pheromones were not processed.

Sows were stimulated with natural sex pheromones 5 days before weaning and the next 15 days daily. The dose of drugs was 0.5 ml / goal. When they showed sexual hunting in sows, they were artificially inseminated. Subsequently, the number of farrowing sows and the number of piglets obtained were taken into account.

The results of the third scientific and production experiment showed that within 20 days after weaning of piglets among animals subjected to stimulation with the prepared preparation of natural sex boar pheromones, sexual hunting was detected in 27 (90%) sows. After stimulation with the prototype drug, sexual hunting was observed in 25 (83%) sows, and among control animals - in 19 (63%). Moreover, more piglets (235 goals) were obtained from sows of the 1st experimental group than from sows of 2 (196 goals) and 3 (144 goals) groups (Table 4).

Thus, the inventive method of obtaining a preparation of natural sex boar pheromones allows to obtain a preparation having high biological activity and a longer shelf life.

Table 1 The results of biological testing of the manufactured preparation of natural sex boar pheromones and the prototype drug Object under investigation Biotest Results (%) Manufactured drug shelf life 12 months 89.5 shelf life 24 months 88.7 Prototype drug shelf life 12 months 78.8 shelf life 24 months 60.0 Urine of a Mature Boar 68.5 Extract from celery root root 57.8 Extract from the root of the parsnip 54.0 Compound feed broth 40,0 Distilled water 4.7

table 2 The onset of puberty in repair pigs subjected to stimulation with preparations of natural sexual boar pheromones Group n There was puberty in pigs within a day after stimulation day 1-5 6-10 11-15 16-20 21-25 26-30 Total 1 (manufactured drug) 35 eleven 10 5 5 - - 31 (89%) 2 (prototype drug) 35 8 8 6 3 3 28 (80%) 3 (control) 35 - 2 - 5 5 four 16 (46%)

Table 3 The onset of puberty in repair pigs with delayed maturity, subjected to stimulation with preparations of natural sexual boar pheromones Group n There was puberty in pigs within a day after stimulation day 1-5 6-10 11-15 16-20 21-25 26-30 Total 1 (manufactured drug) fifteen four 5 2 - - - 11 (73%) 2 (prototype drug) fifteen four four - one - - 9 (67%) 3 (control) fifteen - - 2 - one - 3 (20%)

Table 4 The influence of natural reproductive boar pheromones on the onset of sexual hunting in sows and the results of insemination Group n Came into the hunt for 20 days after weaning piglets Farrowing sows Piglets received quantity % quantity % Total multiple pregnancy 1 (manufactured drug) thirty 27 90 24 89 235 9.8 2 (prototype drug) thirty 25 83 twenty 80 196 9.8 3 (control) thirty 19 63 16 84 144 9.0

Claims (1)

A method of obtaining a preparation of natural sexual boar pheromones, including grinding testes, homogenizing them, adding the protosubtilin enzyme, incubating 1.0-1.5% of the total weight of the feedstock, incubating for 50-60 minutes at 37-39 ° C, centrifuging, mixing with urine in a ratio of 1.0: 1.0-1.5, characterized in that the root celery root and seed parsnip root in a ratio of 1.0: 0.25: 0.25 are added to the initial mass of testes, the resulting mass after adding urine is squeezed in a press with cells of 0.7 × 0.7 mm, and the liquid is sublimi digging.