RU2430731C1 - Polyfunctional enterosorbent - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention concerns veterinary science. A polyfunctional enterosorbent contains schungite-containing minerals comprising silicon dioxide 15.0-70.0 wt %, with an average median particle size 15.0*10^-6 m.

EFFECT: invention provides higher efficacy on a wide spectrum of toxic substances, including mycotoxines, nitrates, nitrites, heavy metal salts, and also as an antibacterial and antioxidant agent.

14 tbl, 14 ex

Description

The invention relates to the use of natural sorbents, in particular to a new enterosorbent based on schungite-containing natural mineral raw materials with high sorption properties in relation to a wide range of toxic substances, as well as exhibiting the properties of an antibacterial agent, and can be used in veterinary medicine to increase weight gain, preserve livestock and resistance of farm animals and poultry, as well as for the prevention and treatment of gastrointestinal diseases of agricultural Landing animals and birds.

Lignin-based enterosorbents are known, for example (RU 2234931, 2002.07.29), (RU 2084236, 1996.02.20). They have several disadvantages, which are the presence of side effects of the use of lignin and lignin-containing products.

A method for producing an enterosorbent based on activated carbon fibers, treated with a binder, which uses starch gel (RU 2027437, 1995.01.27), is described. The disadvantage of this development is the use of particles of carbon fibers having a length at the level of millimeters, such particles have a sharply reduced sorption capacity in comparison with particles having nanoscale sizes. The effectiveness of carbon as a sorbent almost completely disappears if these carbon particles are coated with starch. When it enters the gastrointestinal tract, starch dissolves, while most of the components are desorbed from starch.

The carbon-mineral sorbent SUMS-1 (RU 2143946, 2001.01.10) is known for various purposes, including for medicine as a hemosorbent, enterosorbent, etc., one of the disadvantages of which is the content of toxic and carcinogenic substances in carbon after burning.

Of great importance are sorbents based on silicon-containing natural mineral raw materials. Known enterosorbent-shivyrtuin made from silica-containing raw materials containing a total of 60-80% clinoptilolite and montmorillonite, as well as impurities of quartz and biotite (RU 2122868, 10.12.1998). Enterosorbent exhibits sorption activity against microorganisms, weakens the diffusion of toxins, and stimulates the migration of leukocytes into the wound. Of the identified deficiencies, absorption of vitamin D is essential for surgery, in addition, shivirtuin promotes the growth of Candida mushrooms in the lumen of the digestive tract, which is offset by the addition of appropriate drugs (nystatin or levorin).

There is a description of a sorbent based on silica-containing mineral raw materials containing sedimentary opal-cristobalite rock containing not more than 15 wt.% Clay minerals of the montmorillonite group and not more than 10 wt.% Minerals of the mica group (RU 2319488, 03.20.2008). The patent applicant established that the content of clays of the montmorillonite group in the enterosorbent should not exceed 15 wt.%, Since the minerals of this group have a high swelling in liquids, and when their content is above 15%, difficulties arise in enterosorption through a probe. The mineral content of the mica group should be limited to 10 wt.%, Since with a standard degree of grinding of the raw material (0.1-1 mm), the edges of the mica plates are sharp enough, which can cause various problems in the body.

Closest to the claimed invention and, therefore, selected as a prototype is a multifunctional sorbent Polysorb VP, used in veterinary medicine, which is a highly pure (at least 92% of the basic substance) highly dispersed silica, which is produced in the Chelyabinsk region [Reference. Veterinary drugs in Russia. T.1 / Ed. Klenova I.F. and others // LLC "Selkhozizdat", Moscow - 2004. - S.526-527]. Particle size not more than 0,090 mm, specific surface not less than 150 m ² / g. In the lumen of the gastrointestinal tract, Polysorb VP binds and removes endogenous and exogenous toxic substances of various nature from the body, including microorganisms and microbial toxins, antigens, feed allergens, salts of heavy metals, radionucleotides.

The aim of this invention is the creation of a new highly effective multifunctional enterosorbent, expanding the range of known enterosorbents and having high sorption properties against a wide range of toxic substances, as well as exhibiting antibacterial and antioxidant properties.

The technical result of the invention is the high sorbing capacity in vivo in relation to a wide range of toxic substances of the proposed enterosorbent, which also exhibits antibacterial and antioxidant properties.

The technical result is achieved by using the claimed enterosorbent from natural schungite-containing mineral raw materials with a carbon content of from 3.998% to 55.1% of the mass. and silicon oxide from 15.1% to 70% of the mass, with an average median particle size of enterosorbent of 15.0 * 10 ^-6 m, as an additive to the diet of farm animals and birds (broiler chickens, laying hens, pigs and large cattle) at the rate of 0.1-1.0% of the mass. by weight of feed.

Shungite-containing mineral raw materials are Precambrian rocks with a silicate mineral base, saturated with carbon (schungite) material in a non-crystalline state, subdivided into low-carbon shungite-containing (up to 5% carbon), medium-carbon shungite (5-25% carbon) and high-carbon (25–80 carbon) % carbon) (Sidorenko S.A., Sidorenko A.V. Organic substance in sedimentary-metamorphic rocks of Precambrian. M., 1975).

The inventive enterosorbent is prepared as follows.

The prepared ore of schungite-containing mineral raw materials of various origin is crushed, a fraction of no more than 45.0 * 10 ^-6 m is separated, heated at a temperature of 100 to 200 ° C for 10-60 minutes and cooled to ambient temperature.

The finished product is a fine powder with an average median size of 15.0 * 10 ^-6 m, the physico-chemical properties of which are shown in table 1.

Grinding of mineral raw materials to a finer state leads to agglomeration of particles and, as a consequence, a change in the structure and properties of the enterosorbent. Larger particles of enterobenthos, although they exhibit sorption properties, are inferior to those claimed in terms of degree of effectiveness.

Table 1 Physico-chemical properties of the claimed enterosorbent Name of indicator Value Appearance Dark gray to black powder Mass fraction of carbon in the free state,% mass. 4.0-80.0 Mass fraction of silicon dioxide,% of the mass. 15.0-70.0 The content of water-soluble substances,% 0.1-5 Bulk density, kg / m ³ 500-800 True density kg / m ³ 2500-2550 Loss on ignition,% 0.1-5 pH of an aqueous extract 6-9 DBP Adsorption * cm ^3/100 g according to GOST 7885-86 30-35 Adsorption of methylene blue *, g / 100 g according to GOST 4453-74 fifteen Mass fraction of sieve residue after wet sieving No. 0045,%, less 0.08

Belongs to hazard class 4 according to GOST 12.1.001-76.

The introduction of enterosorbent in the diet of farm animals and poultry (broiler chickens, laying hens, pigs and cattle) at the rate of 0.1-1.0% of the mass. the weight of the feed has a positive effect on the physiological parameters of animals.

The following examples are presented in order to more fully illustrate the claimed invention.

Example 1

Shungite-containing mineral raw materials from the Zazhoginsky deposit in an amount of 250 kg with a carbon content of 25.3% of the mass. and silicon oxide 37.2% of the mass. less than 5 * 10 ^-3 m in size after preliminary removal of moisture and debris is crushed in a mill for one hour. The product of grinding quantitatively from the mill is fed into the air classifier by a stream of dried air for separation, particles of more than 45.0 * 10 ^-6 m in the amount of 5 kg are returned to the mill for regrinding. The finely dispersed fraction formed in an amount of 245 kg per hour is not larger than 45.0 * 10 ^-6 m and is fed into a thermal furnace, which ensures uniform heating of mineral raw materials at a temperature of 200 ° C for 10 minutes, after which the product is cooled in an heat exchanger to ambient temperature Wednesday.

Example 1A

Example 1A is carried out under the conditions of example 1, but after the classifier, the grinding product is selected and fed into the thermal furnace, the particle size of which is not more than 100 * 10 ^-6 m.

Example 1B

Example 1B is carried out under the conditions of example 1, but after the classifier, the grinding product is selected and fed into the thermal furnace, the particle size of which is no more than 10 * 10 ^-6 m.

During heat treatment, a noticeable agglomeration of particles occurs with the formation of large pieces of irregular shape of an increased size.

The sizes of agglomerated particles are less than 300 * 10 ^-6 m.

Example 2

Example 2 is carried out under the conditions of example 1, but using schungite-containing mineral raw materials from the Tekeli-Koksu zone with a carbon content of 3.998% wt., Silicon oxide 69.93% wt. and heat treatment of the separated fraction is carried out for 60 minutes at a temperature of 100 ° C.

Example 2A

Example 2A is carried out under the conditions of example 2, but after the classifier, the grinding product is selected and fed into the thermal furnace, the particle size of which is not more than 100 * 10 ^-6 m.

Example 3

Example 3 is carried out under the conditions of example 1, but using schungite-containing mineral raw materials of the Zazhoginsky field with a carbon content of 55.1% by mass, silicon oxide 15.1% by mass.

Example 3A

Example 3A is carried out under the conditions of example 3, but after the classifier, the grinding product is selected and fed into the thermal furnace, the particle size of which is not more than 100 * 10 ^-6 m.

Example 4

The use of enterosorbent for broiler chickens

The experiment is carried out on broilers of cross "ISA" with the cellular method of keeping birds. In the daily age form the control and experimental groups.

The enterosorbent obtained in example 1 is introduced into the feed mixture of the experimental group by gradual mixing in silos in the amount of 0.1% of the diet or 1 g per 1 kg of feed. The feeding and rearing period of broiler chickens is 42 days.

The control group was fed food in the same volume and the same composition as the experimental one, but without the addition of a composite enterosorbent to the diet.

To assess the livestock status of the experimental and control groups, the following indicators are taken into account: weight - by weighing the young and removing the average live weight; daily safety control; immunobiochemical blood counts of chickens during slaughter.

The results of the analysis of weighing indicators are shown in table 2.

table 2 The dynamics of clinical and physiological parameters Indicators Groups of chickens experienced control The number of goals in the group 42000 42000 Live weight of chickens, g - at the moment of feeding (i.e. daily age) 37.3 ± 0.12 37.3 ± 0.13 - in 7 days 140 ± 1.0 123 ± 1.0 - after 14 days 396 ± 1.6 376 ± 1.6 - after 28 days 1150 ± 2.4 998.5 ± 2.2 - after 42 days 2000 1700 Preservation,% 99,2 97.3 The average daily gain, g 46.7 39.5 Feed costs for the growing period per 1 kg of growth, kg (conversion) 1,58 1.70

As can be seen from the data presented, the indicator of live weight of chickens in the experimental group is greater compared to the control within a week from the start of feeding and continues to be large throughout the entire period of feeding. The safety index of broiler chickens was increased (99.2% compared with the control 97.3%), as well as the average daily gain (46.7 g for the experimental group, 39.5 g for the control). At the same time, feed costs for the growing period were reduced by 1 kg of growth (conversion) - 1.58 kg for the experimental group versus 1.70 kg for the control.

The results of the analysis of the biochemical composition of the blood of broiler chickens during slaughter are shown in table 3.

Table 3 The biochemical composition of the blood of broiler chickens before slaughter Indicators Groups of chickens With the use of enterosorbent Without the use of enterosorbent Biochemical parameters Total protein, g% 4.8 ± 0.3 4.3 ± 0.25 Reserve alkalinity, vol.%, WITH 45.6 ± 0.06 39.6 ± 0.06 Calcium, mg% 17.9 ± 0.8 16.6 ± 0.9 Phosphorus, mg% 4.9 ± 0.5 4.2 ± 0.4 Indicators of natural resistance Bactericidal activity of blood serum,% 52.63 ± 2.13 47.80 ± 2.14 Lysozyme activity of blood serum,% 18.71 ± 0.56 16.90 ± 1.15 Phagocytic activity,% 49.33 ± 2.25 46.14 ± 2.27 Immunoglobulins, units 5.02 ± 0.56 4.07 ± 0.56

The data in table 3 show that the difference in the biochemical composition of blood and the natural resistance of the control and experimental groups proves that the claimed feed additive stimulates the natural resistance of the body of chickens.

Thus, it was shown that the introduction of enterosorbent at the rate of 0.5% per kg of feed as an additive to the diet of broiler chickens has a tangible effect to stimulate growth energy, increase the safety and resistance of birds.

Example 5

The use of enterosorbent for laying hens.

The experiment is carried out with the cellular method of keeping poultry cross-country Hi-line. Form the control and experimental groups.

The enterosorbent obtained in example 2, is introduced into the feed mixture of the experimental group by gradual mixing in silos in the amount of 0.5% of the diet. The observation period for laying hens is 150 days.

The control group is fed with food in the same volume and the same composition as the experimental one, but without the addition of enterosorbent to the diet.

To assess the livestock of the experimental and control groups, the following indicators are taken into account: egg production, safety and weight gain of laying hens, which are shown in table 4.

Table 4 Indicators of egg production, preservation and weight gain of laying hens when observed for 150 days Indicators Group of laying hens experienced control Number of goals 21000 21000 Egg production,%, pcs. eggs per day for the number of days - after 10 days - 0.79 ± 0.07 0.78 ± 0.07 - after 25 days 0.85 ± 0.09 0.80 ± 0.09

- after 50 days 0.92 ± 0.05 0.82 ± 0.05 - after 75 days 0.95 ± 0.04 0.85 ± 0.04 The average gain of laying hens, g, before feeding 1202 1185 after feeding (after 150 days) 1305 1250 Preservation for 150 days,% 98.3 95.2

As can be seen from the data presented, the egg production rate in the experimental group compared to the control increased after 10 days of adding the declared enterosorbent to the diet (0.79 versus 0.78), continued to exceed the control throughout the experiment and after 75 days reached a difference of 10 %

The safety index of laying hens was increased (98.3% compared with the control 95.2%), as well as the egg production rate.

Example 6

The use of enterosorbent for pigs

The experiment is conducted on large white pigs of 100-110 days old. The enterosorbent obtained in example 3 is introduced into the feed mixture of the experimental group by gradual mixing in silos in an amount of 0.2% of the diet. The control group is fed with food in the same volume and the same composition as the experimental one, but without the addition of enterosorbent to the diet.

Indicators of weight gain are shown in table 6.

Table 6 Pig Weight Gain Indicators Control group Experienced group The number of animals in the group, goals thirty thirty Average weight before entry, kg 31.3 ± 0.6 32.5 ± 0.2 Multiplicity of input per day 3 times 3 times Average weight 30 days after application, kg 44.2 ± 0.3 47.6 ± 0.28 The average daily gain, kg 0.432 0.503

As can be seen from the data presented, at the time of the start of the tests, the average weight of the experimental and control groups were almost the same, and after 30 days of use of enterosorbent, the average weight in the experimental group was 3.4 kg more than the control (47.6 ± 0, 28 kg for the experimental group, 44.2 ± 0.3 kg for the control), the average daily gain for the experimental group is 71 g more than in the control (0.503 and 0.432 kg, respectively).

Example 7

Study of the effectiveness of enterosorbent for the treatment of calf diarrhea

The experiment is carried out on calves aged 3-12 days.

Sick animals on the basis of analogues are divided into 2 groups of 20 calves in each. The calves of the experimental group for the treatment of diarrhea use enterosorbent obtained in example 2, at the rate of 1.0% of the mass. by the weight of the feed, the drug was not given to the control group, only the feed.

Animals of both the experimental and control groups are monitored daily, taking into account the clinical condition, appetite, and timing of recovery. Animals are considered to have recovered, in which the clinical signs of the disease disappeared within 10 days from the start of treatment and no relapses were observed. The results are shown in table 7.

Table 7 Enterosorbent efficacy for calf diarrhea Indicators Without enterosorbent With enterosorbent The number of animals in the group twenty twenty The frequency of administration per day (times) 2 2 Application period (days) - 7 Recovery period (days) - 3 Recover (goal) 17 19 Therapeutic efficacy,% 85% 95%

It was found that the use of enterosorbent in the experimental group caused an improvement in the general condition of the animals with subsequent recovery on the 2-3 day of treatment. The therapeutic efficacy of enterosorbent was 95% compared with the control, where the drug was not asked.

Example 8

Determining the effectiveness of the claimed enterosorbent as a sorbent of mycotoxins from the bird

The experiment is carried out in a poultry farm, in which an outbreak of mycotoxicosis occurred - the feed at the time of the experiment already contained mycotoxins formed under conditions of improper storage.

Researches are carried out on broilers of cross "ISA". At the age of 24 days, the control and experimental groups of 42,000 animals each are formed.

In the diet of the broilers of the experimental group, the enterosorbent obtained in Example 1 is introduced from the daily age until slaughter in the amount of 0.1% per kg of feed as an additive to the diet. Enterosorbent is introduced into the feed mixture in the feed workshops of poultry farms by gradual mixing in silos.

To monitor the dynamics of changes in the content of mycotoxins in poultry tissues (liver, stomach and its contents, as well as feces), during colonization and slaughter, samples are taken from the control and experimental groups by irretrievable removal of 20 birds from each group. The determination of the content of mycotoxins corresponds to the normative and technical documentation (for aflotoxin B1 - GOST 30711-2001, for T2-toxin - Guidelines for determining the content of T-2 toxin in biological tissues and extracts (Approved December 27, 1989) (Antonov B.I. ., Yakovleva TF, Deryabina VI Laboratory research in veterinary medicine: biochemical and mycological. - M .: Agropromizdat, 1991, p.146), for zearalenone F-2 - Method for determination of mycotoxin F-2 (zearalenone) in the organs and tissues of animals (Approved May 31, 1987) (Antonov B.I., Yakovl va T.F., Deryabina V.I. Laboratory tests in veterinary medicine: biochemical and mycological. - M .: Agropromizdat, 1991, p. 151), for ochratoxin - Method for determination of ochratoxin A in animal organs and tissues (Approved April 10, 1984 g.) (Antonov B.I., Yakovleva T.F., Deryabina V.I. Laboratory studies in veterinary medicine: biochemical and mycological. - M.: Agropromizdat, 1991, p. 153).

The obtained averaged (from 20 goals) results of changes in the content of mycotoxins are presented in table 5.

The content of various mycotoxins in the samples taken from the control and experimental groups during settlement was approximately the same, this can be explained by the fact that the bird of both groups was taken from the same brood; feed at the beginning of the experiments were also identical.

The data in table 8 show that the content of mycotoxins in the internal organs of the control group of chickens is significantly higher compared to the experimental group that received enterosorbent as a feed additive. An increase in mycotoxins in the contents of the stomach and feces of the experimental group clearly shows that mineral raw materials bind toxins in the stomach and freely remove them from the body of birds. The above results clearly indicate that the introduction of enterosorbent in the feed of broiler chickens has a significant effect in removing mycotoxins from the body.

Example 9

Determining the effectiveness of the claimed enterosorbent as a sorbent of nitrates and nitrites from the bird

The experiment is carried out on broilers of cross "ISA". At the daily age, the control and experimental groups (n = 40) are formed on the principle of analogue pairs.

Both groups are seeded with sodium nitrate (NaNO ₃ ) at the rate of 50 mg NaNO ₃ per kg of feed three times during the life of the bird.

Starting from the settlement, the experimental group is fed the declared enterosorbent obtained according to Example 2, which is added to the feed mixture by the method of stepwise mixing at the rate of 0.5% per kg of feed.

At the slaughter (after 42 days), samples of the liver and muscle tissue are taken. Sampling methods and their preparation for testing comply with the normative technical documentation (GOST 9792-73), they are carried out in accordance with the methods that have passed state certification GOST 29300-92 (ISO 3091-75) for nitrate, GOST 29299-92 (ISO 2918- 75) for nitrite.

The results averaged over 40 goals for the control and experimental groups are presented in table 9.

Table 9 The content of nitrates and nitrites in the organs of broiler chickens No. p / p Name of organ sample Nitrite Nitrates Control group (average results obtained from 40 goals), mg / kg one Muscle tissue 7 ± 0.34 Not found 2 Liver 22.04 ± 0.47 Not found Experimental group (average results obtained from 40 goals) mg / kg one Muscle tissue 3.56 ± 0.56 *** Not found 2 Liver 5.32 ± 0.17 *** Not found Note: Data are reliable in comparison with the control group: * - P≤0.05, ** - P≤0.01, *** - P≤0.001

The data in table 9 show that the content of nitrates and nitrites in the experimental group is significantly lower than in the control, the introduction of enterosorbent in the diet of broiler chickens allows to reduce the content: in muscle tissue: nitrite by almost 2 times, in the liver - by 4 times, nitrate content in both groups was not found. This is due to the fact that nitrates are unstable in birds and quickly turn into nitrites.

Example 10

Determining the effectiveness of the claimed enterosorbent as a sorbent of heavy metals from the bird

The experiment is carried out on broilers of cross "ISA". In the daily age, a control and experimental group of 20 goals each is formed.

Both groups are subjected to simultaneous seeding with lead acetate at a dose of 30 mg per 1 kg of feed and cadmium acetate at a dose of 3 mg per 1 kg of feed. The feed is pretreated with 10% acetate solutions of these heavy metals prepared in distilled water. Thus, the seed is produced each time the feed is set.

The experimental group, starting from the settlement, receives the enterosorbent obtained according to example 2, at the rate of 0.2% per kg of feed as an additive to the diet.

After slaughtering the birds (after 42 days), samples of muscle and bone tissue are taken and the contents of lead and cadmium are determined. Sampling methods and their preparation for testing are in accordance with the regulatory and technical documentation (GOST 26929-94); they are carried out according to the Guidelines for atomic absorption methods for the determination of toxic elements in food products 01-19 (47-11-92). Analysis of objects of research for the content of heavy metals is carried out by methods certified by the metrological service of Gosstandart of the Russian Federation: for lead (GOST 26932-86), for cadmium (GOST 26933-86). Studies of poultry organs and tissues for the content of heavy metals are carried out on an atomic absorption spectrometer (GOST 30178-96). The results are presented in table 10.

Table 10 The content of heavy metals in the muscle and bone tissue of broiler chickens of the control and experimental groups Group Indicator White muscles Red muscles Bone Cd (μg / g) Pb (μg / g) Cd (μg / g) Pb (μg / g) Cd (μg / g) Pb (μg / g) control 0.252 ± 0.055 0.481 ± 0.121 0.264 ± 0.054 0.712 ± 0.141 1.247 ± 0.411 5.523 ±, 954 experienced 0.004 ± 0.06 ** 0.125 ± 0.081 ** 0.006 ± 0.040 ** 0.179 ± 0.156 ** 0.087 ± 0.043 ** 0.265 ± 0.139 Note: Data are reliable in comparison with the control group: * - P≤0.05 ** - P≤0.01 *** - P≤0.001

The data in table 10 show that the content of cadmium and lead in the experimental group is significantly lower than in the control, the introduction of the proposed enterosorbent into the diet of broiler chickens allows to reduce the content of heavy metals:

in white muscles: cadmium is 82.6 times, lead is 3.7 times;

in red muscles: cadmium 4 times, lead 4.9 times;

in bone tissue: cadmium 16.6 times, lead - 20.4 times.

Example 11

Determination of antioxidant properties of the claimed enterosorbent

The antioxidant properties of the claimed enterosorbent are investigated by its effect on lipid peroxidation processes. The degree of intensity of the process of lipid peroxidation (LP) is judged by the accumulation of secondary products of lipid peroxidation - malondialdehyde (MDA) in the reaction with 2-thiobarbituric acid [Goncharenko, M.S. Method for assessing lipid peroxidation / M.S. Goncharenko, A.M. Latinova // Laboratory. - 1985. - No. 1. - S.60-61].

The experiments are carried out on white rats. Before setting up experiments, animals are kept in quarantine for two weeks. In the morning, experimental subjects are given one-time complete feed from the nursery of the Federal State Institution of the Federal Center for Toxicological and Radiation Safety of Animals (Kazan). Water is set in decreasing order. The experimental and control groups of animals are formed according to the principle of analogues. During the entire experiment, the animals were in the same conditions of keeping and feeding.

For experimental studies, T-2 toxin is used as one of the most widespread and most dangerous mycotoxins. To assess the effectiveness of the claimed enterosorbent, the Polysorb enterosorbent manufactured by POLISORB, Russia, is used as a comparison drug.

The experiment was conducted on 50 males of nonlinear white rats, animals were divided into 5 groups of 10 each:

Group 1 - biological control, during the entire period of the experiment received "clean" feed, not containing mycotoxins;

Group 2 - animals throughout the entire period of the experiment received "clean" feed, not containing mycotoxins, mixed with the claimed enterosorbent in an amount of 5 g / kg of feed;

Group 3 - the animals were given “toxic” food contaminated with mycotoxin T-2 (1.0 mg / kg feed);

Group 4 - animals received “toxic” food contaminated with T-2 mycotoxin (1.0 mg / kg feed) mixed with Polysorb enterosorbent in an amount of 5 g / kg feed;

5 group - the animals received "toxic" food contaminated with mycotoxin T-2 (1.0 mg / kg feed), mixed with the inventive enterosorbent obtained in example 2, in the amount of 5 g / kg feed.

The inventive sorbent was taken in a maximum dose (5 kg / t of feed, according to the instructions for use). In order that the dose of administration be the same, the compared sorbent Polysorb was taken in the same amount.

The duration of the experiment is 10 days.

The rats' intake of feed and water did not differ in all groups. Survival in all groups was 100%.

On the tenth day, blood was taken for research by decapitation in ten rats from each group. The degree of intensity of the lipid peroxidation (LP) process is judged by the accumulation of secondary lipid peroxidation products - malondialdehyde (MDA) in the reaction with 2-thiobarbituric acid (TBA).

For the study, 1.5 ml of blood with heparin is taken. Centrifugation (5 min, 3000 rpm) separates the plasma from the formed elements of the blood. Next, an analysis is carried out for the content of TBA-active products separately in plasma and in red blood cells. For this, the erythrocyte mass is washed twice with 10 ml of isotonic solution. After washing, the red blood cells are precipitated by centrifugation for 10 min at 3000 rpm. Washed red blood cells hemolize in distilled water in a ratio of 1: 5. For research, 0.3 ml of plasma and 0.3 ml of hemolysate are taken, to which 2.4 ml of H ₂ SO ₄ and 0.3 ml of 10% tungsten-phosphoric acid are added, mixed thoroughly. After 10 minutes, the precipitate formed is separated by centrifugation at 3000 rpm. The supernatant is discarded, and the resulting precipitate is washed twice with 1 ml of water, dissolved in 3 ml of water, 1 ml of a freshly prepared aqueous solution of thiobarbituric acid in acetic acid is added (80 mg of thiobarbituric acid is dissolved in 5 ml of H ₂ O and 5 ml of glacial acetic acid when heated) . The color reaction is carried out in closed chemical tubes at 96 ° C. The reaction is stopped after 60 minutes by cooling the samples in cold water. To remove turbidity, before spectrophotometry, the samples are centrifuged for 15 minutes at 3000 rpm. The optical density is measured on an SF-26 spectrometer at a wavelength of 532 nm in cuvettes with an optical path length of 10 mm. Distilled water is used as a control. The calculation of the amount of malondialdehyde (MDA) is carried out taking into account the dilution according to the formula C = E / έ, where E is the optical density of the sample, έ is the molar instinct coefficient equal to 1.56 × 10 ^-5 mol ^-1 / cm ^-1 , С - the concentration of MDA, expressed in micromoles per 1 ml of plasma and 1 ml of erythrocyte mass. The values of MDA in plasma and red blood cells are summarized.

The research results are presented in table 11.

Table 11 The content of malondialdehyde (× 10 ^-5 mol ^-1 cm ^-1 ) in the blood of rats of groups 1-5 after 10 days Groups Indicator one 0.784 2 0.724 3 0.971 four 0.892 5 0.823

A regular increase in the content of malondialdehyde (MDA) in the blood of rats of 3-5 groups receiving a “toxic” food is recorded in comparison with groups 1 and 2 of the control.

So, in group 3 on the 10th day the increase in the content of MDA compared to the control (1) group was 23.8%, in group 4 - by 13.8%, in group 5 - by 5.0%. At the same time, in the second group, a decrease in the content of MDA by 7.65% was observed. This indicates a more pronounced nonspecific antioxidant effect of enterosorbent in comparison with Polysorb.

The content of malondialdehyde in the blood of rats of groups 4 and 5 is quite close, although in group 5 this value is slightly lower (0.823 × 10 ^-5 mol ^-1 cm ^-1 versus 0.892 × 10 ^-5 mol ^-1 cm ^-1 for the group receiving " Polysorb "), this means that the inventive enterosorbent more actively bound peroxidation products of metabolism than Polysorb.

When studying the adsorption properties of the claimed enterosorbent in relation to various types of microorganisms (Escherichia coli, Salmonella typhimurium, Staphylococcus aureus, Clostridium perfringes), it was found that their significant adsorption was 63.2-91.8%, which is not inferior, and sometimes significantly superior to the adsorption properties of the prototype.

Example 12

The adsorption effect of the claimed enterosorbent in relation to microorganisms is shown on the example of a culture of Escherichia coli.

1 day before the start of the experiment, the culture of Escherichia coli is sown on beveled meat-peptone agar (MPA). Crops are placed in a thermostat for 24 hours at a temperature of 37 ° C. After 24 hours, the cultures are washed off with sterile distilled water, filtered through a sterile filter and a working suspension of microbes is prepared, standardized to an optical density standard of 2 units. At the same time, a working suspension is prepared with the inventive enterosorbent obtained in Example 1. For this, a sterile isotonic solution of 4.5 ml is poured into a sterile tube and 0.5 ml of a working suspension of culture and 25 mg of sterile enterosorbent are added (the amount of enterosorbent used for the experiment is determined by the maximum recommended dose enterosorbent - 5 g / kg feed; to maintain this ratio, the calculation was - 5 g / l sterile isotonic solution). A working suspension with the inventive enterosorbent is placed in the apparatus for shaking samples for 1.5 hours. Next, serial dilutions of this mixture are carried out 1:10, 1: 100, 1: 1000, 1: 10000, of which they are plated on MPA in Petri dishes (three crops for each dilution) from a test tube using the 0.1 ml lawn method the resulting dilutions. The cups were placed in a thermostat for 24 hours at a temperature of 37 ° C.

In parallel, they put controls, i.e. perform the same actions for:

- tubes with a working suspension of microbes without the inventive enterosorbent (basic control, in comparison with which adsorption is calculated);

- test tubes with an isotonic solution and the inventive enterosorbent without microbes (sterility control to ensure the reliability of the results that no other microbes were introduced with water or with the inventive enterosorbent);

- tubes with 4.5 ml of an isotonic solution, 0.5 ml of a working suspension of culture and 25 mg of a sterile enterosorbent prototype.

After 24 hours, the number of colonies grown in three cups is visually counted, the results of the highest dilution with the least number of colonies grown are taken to calculate the CFU, and the average result is output - this figure was then multiplied by the degree of dilution, the obtained number was compared with control crops and the difference in numbers was calculated as adsorption and recounted in percent. Studies were carried out 3 times with stat. processing.

After 24 hours, the number of colonies grown in three dishes is visually counted, the highest dilution results with the least number of colonies grown (1: 10,000) were taken to calculate KOE, the average result was output, which was multiplied by the dilution ratio (10 ⁵ ), and the resulting figure was compared with control crops.

The difference is considered as adsorption and converted into percentages. Studies are carried out three times using statistical processing.

The results are presented in table 12.

Table 12 The adsorption effect of the claimed enterosorbent in relation to the culture of Escherichia coli, KOE / ml (× 10 ⁵ ) KOE / ml The indicator of adsorption activity,% The control fifty Polysorb (prototype) 17 66 Invention fifteen 70

According to table 12, it can be seen that the adsorption with respect to Escherichia coli by the claimed enterosorbent was 70%, and Polysorb - 66%, i.e. the claimed enterosorbent exhibits better adsorption properties against the pathogenic strain of Escherichia coli compared to the prototype.

Example 13

Tests of enterosorbents are carried out using the products obtained in examples 1A, 1B, 2A, 3A, 4A, similarly to the description given in example 12.

The test results are shown in table 13.

Table 13 The adsorption effect of the claimed enterosorbent in relation to the culture of Escherichia coli, KOE / ml (× 10 ⁵ ) KOE / ml The indicator of adsorption activity,% The control fifty Polysorb (prototype) 17 66 Example 1 fifteen 70 Example 1A 21 58 Example 1B 19 62 Example 2A 19 62 Example 3A twenty 60

According to table 13, it is seen that the adsorption with respect to Escherichia coli by the enterosorbents obtained in Examples 1A, 1B, 2A, 3A was lower. From the above data it follows that the particle size is a significant factor determining the adsorption properties with respect to the pathogenic strain of Escherichia coli, and exhibits the best properties when the maximum particle size lies in the range up to 45 * 10 ^-6 m and the average statistical median size is 15 * 10 ^-6 m

Example 14

Determining the effectiveness of the claimed enterosorbent as a sorbent of mycotoxins from the bird

Tests of enterosorbents in this example were carried out using enterosorbents obtained according to examples 1A, 1B, similarly to the description given in example 8. Analogously to example 8, the enterosorbent obtained in example 1 was added to the diet of broilers of the experimental group from the day old to the slaughter, in the amount of 0, 1% per kg of feed as an additive to the diet. Sorption efficiency was evaluated by the content of one of the mycotoxins Aflotoxin B1 was checked using enterosorbents obtained according to the procedure described in examples 1, 1A and 1B. The results are shown in table 14.

Thus, a multifunctional enterosorbent based on shungite-containing mineral raw materials is declared, which has high sorption properties against a wide range of toxic substances, including mycotoxins, nitrates, nitrites, salts of heavy metals, as well as showing the properties of an antibacterial agent and antioxidant effect.

The inclusion of this enterosorbent in the diet of farm animals and birds has a positive effect on the increase in weight gain, egg production, preservation, and resistance. The obtained results give reason to recommend the inventive enterosorbent for the prevention and complex therapy of toxicosis and diseases of the gastrointestinal tract of various etiologies in farm animals and birds.

Claims (1)

A multifunctional enterosorbent based on natural silicon-containing mineral raw materials, characterized in that, as a natural silicon-containing mineral raw material, it contains schungite-containing mineral raw materials with a silica content of 15.0-70.0 wt.%, And the average median particle size of the enterosorbent is 15.0 10 ^-6 m.