RU2614790C2 - Production method for sterile lyophilised additive from drone larvae and pupae - Google Patents

Abstract

FIELD: food industry.

SUBSTANCE: invention relates to fodder production and veterinary science, in particular, to the method for production of sterile lyophilsed additive from drone larvae and pupae. Additive production method involves collection of drone larvae and pupae, their freezing, storage, defrosting with further homogenization, autoclaving, packaging and freeze drying. Drone larvae and pupae are frozen at the temperature of -(18-20)°C. Homogenization is performed during 1-3 minutes at 3000-4000 rpm. Prior to freeze drying, the vialed homogenate is frozen in a refrigerator with at least 5-row stack which is precooled down to -(44-45)°C. Drone pupae are collected until 20 days of age. Prior to packaging, distilled water is added to the homogenate at the ratio 1:1, and sucrose-gelatin medium (10:2) is added to the obtained suspension at the ratio of 2 parts of the obtained suspension - 1 part of the said medium with further repeated homogenization at a colloid mill and autoclaving follows at +(118-122)°C during 43-45 minutes. Freeze drying takes place in vacuum with the total drying time being 98-100 hours.

EFFECT: invention ensures production of fodder additive with its feeding providing for the increase of animal mating activity, increase of body resistance and growth of live weight.

1 dwg, 1 tbl, 5 ex

Description

FIELD OF THE INVENTION

The invention relates to veterinary medicine, in particular to methods for sterile lyophilized additives.

State of the art

A known method of preparation of a preparation for stimulating the organism of animals from drones larvae, including collecting larvae, their homogenization with the addition of a stabilizing medium, while before homogenizing drones larvae are frozen at a temperature of (-4; -5 ° C) for 12-14 hours, then thawing to a temperature of + 18 ° C and homogenized for 3-5 minutes, while a stabilizing medium using a 5% glucose solution in a ratio of 1:10 with the addition of a 5% solution of chloroform or 2% salicylic acid, followed by sublimation, and blimate is carried out in a vacuum chamber at a temperature of shelf heating to 30 ° -45 ° C, while reaching a temperature of 20-22 ° C, the temperature of the shelves is reduced to 25 ° C and drying is carried out for 22-24 hours (see. "Preparation of drones larvae ", VI Lebedev, MA Legovich, Research Institute of beekeeping, J. Beekeeping, No. 3, 2003, S. 52-54).

The disadvantage of this method is the short storage time of the drug.

There is a method and means of improving the health of athletes, including the appointment of a complex drug Apimikroelfit containing dry extracts of Eleutherococcus, licorice root, rosehip, tincture of propolis, estifan, silymar and honey, with the following ratios of components, wt. %: dry Eleutherococcus extract 0.6; dry licorice root extract 0.3; dry rosehip extract 0.3; estifan 0.3; silymar 0.3; propolis tincture 0.3; natural honey the rest. The tool is administered in the pre-competition and competition period for 7-10 days, 1 teaspoon diluted in 0.5 tbsp. water, 2-3 times a day, only 150-200 g per course, and carry out 2-3 courses with an interval of 7-10 days (see Pat. RF No. 2104013, CL A61K 35/64, A61K 35/78, published: 02/10/1998).

The disadvantage of this method is the complicated method of preparation, the high cost of the components and the long course of use.

The closest in technical essence and the achieved positive effect and adopted by the authors for the prototype is a method of preparing a lyophilized biologically active additive for animals, which includes collecting 9-10 day old drones larvae, freezing them, storing them before use, followed by homogenization and packaging, in addition use pumpkin and distilled water. The pumpkin is cleaned, cut into small pieces, frozen at a temperature of - (18-20) ° C for 22-24 hours, then thawed to a temperature of + (17-18) ° C for 9-11 minutes, homogenized, followed by connection with homogenate of drones larvae, and drones larvae are frozen at a temperature of - (18-20) ° С, and drones and pumpkin larvae are homogenized to particle sizes of 0.5-1.0 mm. Next, before packing, homogenate is mixed in a ratio of 1 part of drones larvae 4 parts of pumpkin and 5 parts of distilled water, followed by freeze drying and sublimation in compliance with certain parameters of these operations (see US Pat. RF No. 2435433, class A23K 1 / 00, published on December 10, 2011).

The disadvantage of this method is that this drug is unsterile, does not have sufficiently high stimulating properties.

Disclosure of invention

The objective of the invention is to develop a method for producing a sterile feed additive from larvae and pupae of drones of bees to increase the sexual activity of sheep undergoing protozoal diseases (anaplasmosis).

The technical result that can be achieved using the present invention is to increase the sexual activity of animals and increase the body's resistance, increase body weight.

The technical result is achieved using the method of obtaining a sterile lyophilized additive from larvae and pupae of drones, including collecting larvae and pupae of drones up to 20 days of age, their freezing, storage (not more than 10 months), thawing, followed by homogenization, autoclaving, packing and freeze drying moreover, the larvae and drones pupae are frozen at a temperature of - (18-20) ° С, and homogenization is carried out for 1-3 minutes at 3-4 thousand / revolutions per minute. In this case, before packing, distilled water is added to the homogenate in a ratio of 1: 1 and a sucrose-gelatin medium in the ratio of 2 parts of the suspension obtained is 1 part of the medium, followed by repeated homogenization in a colloid mill and placed in an autoclave at + (118-122) ° С for 43-45 minutes. In this case, before freeze drying, the homogenate packaged in bottles is frozen in the refrigerator with at least a five-shelf shelf pre-cooled to - (44-45) ° С, and after loading, the freezing temperature is set to (19-20) ° С and maintain this temperature for 5 hours, and then set the temperature to (44-45) ° C, which is maintained for 9 hours, with a total freezing time of 13-14 hours, followed by reinstallation of the shelf with bottles in a sublimation chamber with a set heating temperature + ( 20-21) ° C, s a subsequent increase in the heating temperature by 5 ° C from + (20) ° C to + (30) ° C, maintaining this temperature until the end of drying, and drying is carried out for 44-45 hours after reaching a temperature of + 22 ° C in the product, and freeze drying is carried out in vacuum with a depth of (≥8 mV) with a total drying time of 98-100 hours.

The result of increased sexual activity is achieved by introducing into the additive larvae and pupae of drones, which, due to their constituent components, stimulate the central mechanisms of regulation of androgen formation, contribute to the restoration of impaired sexual function and increase sexual desire. These components contribute to the accelerated restoration of blood biochemical characteristics, improve the general condition and appetite (Nemtsov S.V., Zueva O.Yu., Khismatullin M.R., Larikov V.V., Varlamov V.P., 2001; Mashenkov O. N., 2005).

The dried larvae and pupae of bees contain 28 amino acids, including 9 essential amino acids (see “Biological and nutritional value of Bilar powders”, A.I. Cherkasov, I.A. Prokhod, J. “Beekeeping”, No. 2, 2005, p. 50-51). Drones larvae contain a high level of carbohydrates, minerals, as well as biologically active substances: a large number of functional groups of enzymes, as well as testosteroid hormones (see “Using drone brood for cosmetic purposes”, A. G. Bachinsky, A. N. Decina, J. "Beekeeping", No. 5, 1998, p. 54) of progesterone extradiol.

The essence of the preparation of sterile lyophilized additives from the young phases of the drones

The drug is made from larvae and pupae of drones.

In the apiary, larvae and pupae of drones are collected up to 20 days of age together with drone milk in a clean plastic dish, then placed in a freezer at a temperature of - (18-20) ° С (storage period should not exceed 10 months). Before use, the biological material is thawed at room temperature and homogenized in a blender for 1-3 minutes at 3-4 thousand revolutions / minute. An equal volume of distilled water (1: 1 ratio) is added to the homogenate. A sucrose-gelatin medium (10: 2) is added to the resulting suspension in a ratio of 2 parts of the suspension to 1 part of the medium. The resulting mixture is homogenized in a colloid mill, placed in an autoclave and autoclaved at + (118-122) ° C for 43-45 minutes. The homogenate is packaged in 100 cm ³ vials of 40 cm ³ and loaded into the HC 700/50 refrigerator with a five-shelf shelf pre-cooled to - (44-45) ° С. After loading set the freezing temperature - (19-20) ° C and maintain this temperature for 5 hours. Then set the temperature - (44-45) ° C, which is maintained for 9 hours with a total freezing time of 14 hours. The shelf with bottles is reinstalled in a sublimation chamber of the LZ-45 type, 1 hour after creating the necessary depth (≥8 mV) in the sublimation chamber, the heating temperature is set to + (20-21) ° С, after 1 hour the heating temperature is increased by 5 ° C to + 25 ° C; after reaching the product temperature of + 25 ° C, the temperature is increased to + 30 ° C, which is maintained until the end of freeze drying. Draining is carried out within 44-45 hours after reaching a temperature of + 22 ° C in the product, with a total drying time of 98-100 hours.

After freeze drying is complete, the vials with the drug are unloaded, corked with stoppers and wrapped in caps.

The implementation of the invention

Examples of specific preparation of a sterile lyophilized supplement from young phases of drones

Example No. 1. In the apiary, larvae and pupae of drones are collected until 20 days of age in clean plastic dishes, then placed in a freezer at -18 ° C and stored until production (but not more than 10 months). Subsequently, they are thawed at room temperature for 6-8 hours and homogenized in a blender at 3 thousand / revolutions per minute for 5 minutes. Distilled water is added to the homogenate in a ratio of 1: 1, mixed, poured into vials with a capacity of 100 cm ³ by 40 cm ³ , and the homogenate in the vials is loaded into a HC 700/50 refrigerator with a five-shelf shelf pre-cooled to -45 ° C. After loading, set the freezing temperature to -20 ° C and maintain this temperature for 5 hours. Then set the temperature to -45 ° C, which is maintained for 9 hours with a total freezing time of 14 hours. The rack with bottles is reinstalled in a sublimation chamber of the LZ-45 type, 1 hour after creating the necessary depth (≥8 mV) in the sublimation chamber, the heating temperature is set to + 20 ° C, after 1 hour the heating temperature is increased by 5 ° C to +25 ° C; after reaching the product temperature of + 25 ° C, the temperature is increased to + 30 ° C, which is maintained until the end of freeze drying. Draining is carried out within 45 hours after reaching a temperature of + 22 ° C in the product, with a total drying time of 100 hours.

After freeze drying is complete, the vials with the drug are unloaded, corked with stoppers and wrapped in caps.

With this method, an incompletely formed tablet was obtained, which quickly crumbled. When diluted with water, a suspension was obtained containing large inclusions. When sowing on liquid and solid nutrient media, active growth of microorganisms was observed.

Example No. 2. The collection and storage of larvae and pupae of drones is carried out similarly to the first example. In contrast to the first example, after homogenization, distilled water in a ratio of 1: 1 is added to the homogenate, and a sucrose-gelatinized medium in a ratio of 10: 2 is added to the resulting suspension. Then it is homogenized twice in a PUCCOLOIDTECHNIK colloid mill of type JV14, poured into 100 cm ³ vials of 40 cm ³ and lyophilized, as in Example 1. After drying, the drug vials are also unloaded, corked and sealed with caps.

In this method, a formed yellow-colored tablet was obtained that easily dissolved in water without large particles. However, when sowing on liquid and dense nutrient media, in the same way as in the first example, active growth of microorganisms was observed.

Example No. 3. The collection, storage and preparation of larvae and pupae is carried out analogously to example 2, but after double homogenization in a colloid mill and filling the bottles, the homogenate is autoclaved at 120 ° C for 45 minutes and subjected to freeze drying in the same mode as in example 2.

In this case, a light yellow tablet was formed, which easily dissolved in water. When inoculated on bacteriological media, the growth of microorganisms was not observed, the drug is sterile.

Example No. 4. The collection, storage and preparation of larvae and pupae is carried out analogously to example 3, but after double homogenization in a colloid mill and filling the bottles, the homogenate is autoclaved at 118 ° C for 50 minutes and subjected to freeze drying in the same mode as in example 3.

In this case, a light yellow tablet was formed, which easily dissolved in water. When inoculated on bacteriological media, the growth of microorganisms was not observed, the drug is sterile.

Example No. 5. The collection, storage and preparation of larvae and pupae is carried out analogously to example 3, but after double homogenization in a colloid mill and filling the bottles, the homogenate is autoclaved at 120 ° C for 120 minutes and subjected to freeze drying in the same mode as in example 3.

In this case, a brown tablet formed, which dissolved in water, but an insignificant amount of a brown precipitate formed. When inoculated on bacteriological growth media of microorganisms, it was observed, the drug is sterile.

Thus, the most optimal are examples No. 3 and No. 4 of the preparation of a sterile lyophilized additive from larvae and pupae of drones, which produce a readily soluble in water and sterile biologically active additive, which was subsequently tested.

The invention in comparison with the prototype and other known technical solutions has the following advantages:

- lyophilized supplement increases the sexual activity of males and females;

- lyophilized supplement helps to increase body resistance and increase body weight;

- does not have an allergic effect;

- has sterility;

- has a long shelf life (2 years).

With a specific application of the developed supplement to rams with reduced sexual activity due to anaplasmosis, an increase in sperm production was noted (table 1).

Analysis of the sperm production data (ejaculate volume, activity, density) showed that at the end of the experiment, ram-producers of the experimental group had an ejaculate volume of 0.8-1.3 ml more than in animals of the control group, sperm activity was higher than in the control group, by 2 points, the concentration of sperm in one milliliter of the rams of the experimental group exceeded the animals of the control group by 2.4 million. At the peak of activity, the rams of the experimental group inseminated more than 20 sheep, and the control group - only 6-10 sheep, come into the hunt (picture 1).

Thus, a positive effect of the lyophilized preparation from bee darvae and pupae of larvae of drones of bees on the qualitative and quantitative characteristics of sperm production in ram-producers during anaplasmosis was established. The obtained results suggest that the drug has a positive effect on spermiogenesis due to the additionally obtained amino acids, vitamins, and trace elements.

Claims (1)

A method of obtaining a sterile lyophilized additive from darling larvae and pupae, including collecting drones larvae and pupae, freezing, storing, thawing them with subsequent homogenization, autoclaving, packing and freeze-drying, while larvae and pupae of drones are frozen at a temperature of - (18-20 ) ° C, and homogenization is carried out for 1-3 minutes at 3-4 thousand / rpm, before freeze drying, freeze the homogenate packaged in bottles in the refrigerator, at least with a 5-shelf shelf, chilled to - (44-45) ° С, drona pupae are collected up to 20 days of age, before packing, distilled water in the ratio 1: 1 is added to the homogenate, and sucrose-gelatin medium (10: 2) is added to the resulting suspension in the ratio 2 parts of the resulting suspension - 1 part of the medium, followed by repeated homogenization in a colloid mill and autoclaved at + (118-122) ° C for 43-45 minutes, freeze drying is carried out in vacuum at a total drying time of 98-100 hours.

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