Polyfunctional enterosorbent

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

 

The invention relates to the field of use of natural sorbents, in particular to a new enterosorbent on the basis of schungite comprising natural mineral raw materials with high sorption properties against a broad range of toxic substances, and also shows the properties of an antibacterial agent, and can be used in veterinary medicine to increase in weight, livability and resistance of farm animals and poultry, as well as for the prevention and treatment of gastrointestinal diseases of agricultural animals and birds.

Known chelators on the basis of lignin, for example (EN 2234931, 2002.07.29), (EN 2084236, 1996.02.20). They have a number of disadvantages, which in the presence of side effects of lignin and lignin products.

Describes a method for enterosorbent-based activated carbon fibers treated with a binder, which is used as a starch gel (EN 2027437, 1995.01.27). The disadvantage of this design is the use of particles of carbon fibers, having a length of at the level of millimeters, such particles have sharply reduced the sorption capacity compared to particles having a nano. The effectiveness of carbon as sorbent almost completely disappears if these carbon particles to cover Brahma the om. When released into the gastrointestinal tract starch dissolves, while simultaneously desorbed from starch, most components.

Known carbonaceous mineral sorbent SUMS-1 (EN 2143946, 2001.01.10) for various purposes, including for medicine as hemosorbent, enterosorbent, and others, one of the drawbacks is the content of toxic and carcinogenic substances in the carbon after burning.

Are important sorbents based on silica-containing mineral raw materials. Known enterosorbent-RIVERTWIN, made of silica-containing raw material containing a total of 60-80% of clinoptilolite and montmorillonite, as well as admixtures of quartz and biotite (EN 2122868, running from 10.12.1998). Enterosorbent shows sorption activity against microorganisms, reduces the diffusion of toxins, stimulates the migration of leukocytes in the wound. Identified significant for surgery absorption of vitamin D, in addition, RIVERTWIN promotes the growth of fungi of the genus Candida in the lumen of the digestive tract, which is offset by the addition of appropriate drugs (nystatin or levorin).

There is a description of the sorbent on the basis of silica-containing mineral raw materials containing sedimentary opal-chalcedony and cristobalite breed, containing not more than 15 wt.% clay minerals of the montmorillonite group and not Bo is her 10 wt.% minerals of the group of micas (EN 2319488, 20.03.2008). By the patent applicant found that the content of the clays of the montmorillonite group in the enterosorbent should not exceed 15 wt.%, because the minerals of this group have a high swelling properties of the fluids, and when the content is higher than 15% having difficulty enterosorption through the probe. The mineral content of the group of micas should be limited to 10 wt.%, as standard the degree of grinding of raw materials (0,1-1 mm) of the edge of the mica plates are sharp enough, which can cause various problems in the body.

Closest to the claimed invention and on this basis, selected as a prototype is a polyfunctional sorbent Polisorb EAP, used in veterinary medicine, which represents a high purity (not less than 92% of the basic substance) highly disperse silica, which is produced in the Chelyabinsk region [Handbook. Veterinary medicines in Russia. T.1 /edited maple IVAN and others // OOO Selhozizdat", Moscow 2004. - S-527]. A particle size of not more than 0,090 mm, a specific surface area of not less than 150 m2/year In the lumen of the gastrointestinal tract Polisorb EAP binds and removes from the body of endogenous and exogenous toxic substances of different nature, including microorganisms and microbial toxins, antigens, food allergens, heavy metals, radionucleotide.

The purpose of this invention which is a new high-performance multifunctional enterosorbent, extends the range of known sorbents with high sorption properties against a broad range of toxic substances, as well as exhibiting antibacterial and antioxidant properties.

The technical result of the invention is a high absorptive capabilities in vivo against a wide range of toxic substances proposed enterosorbent, which also exhibits antibacterial and antioxidant properties.

The technical result is achieved by using the claimed enterosorbent of natural schungite comprising mineral raw materials with carbon content from 3,998% to 55.1% of the mass. and silicon oxide from 15.1% to 70 wt%, with an average median particle size of enterosorbent 15,0*10-6m, as an additive in the diet of feeding of agricultural animals and poultry (broiler chickens, laying hens, pigs and cattle) 0,1-1,0% of the mass. on the weight of the feed.

Schungite comprising mineral raw materials is a Precambrian rocks with silicate mineral Foundation, saturated carbon (shungite) substance in the noncrystalline state, subdivided into low-carbon schungite comprising up to 5% carbon), medium songfile (5-25% carbon and high-carbon shungite (25-80% carbon) (Sidorenko S., Sidorenko A.V. Organizes the e matter in sedimentary-metamorphic rocks of the Precambrian. M., 1975).

Declare enterosorbent obtained as follows.

The prepared ore longitudedegrees mineral raw materials of different origin are crushed, the fraction of no more than 45,0*10-6m is separated, is heated at a temperature of from 100 to 200°C for 10-60 minutes and cooled to ambient temperature.

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

The grinding of mineral raw materials to smaller state leads to agglomeration of the particles and, consequently, changes the structure and properties of enterosorbent. Larger particles enterosorbent, although showing sorption properties, but inferior to the claimed efficiency.

Table 1
Physico-chemical properties of the claimed enterosorbent
Name of indicatorValue
AppearancePowder from dark gray to black color
Mass fraction of carbon in a 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/m3500-800
True density kg/m32500-2550
Loss on ignition, %0.1-5
pH of aqueous extract6-9
Adsorption of dibutyl phthalate*, cm3/100 g according to GOST 7885-8630-35
Adsorption of methylene blue*, g/100 g according to GOST 4453-7415
Mass fraction of residue on the sieve after wet prosea No 0045,%,0,08

Belongs to the 4th class of danger according to GOST 12.1.001-76.

The introduction of enterosorbent in the feeding of farm animals and poultry (broiler chickens, laying hens, pigs and cattle) 0,1-1,0% of the mass. on the weight of feed has a positive effect on physiological parameters of animals.

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

p> Example 1

Schungite comprising mineral raw materials of the zazhoginsky deposits in the 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*10-3m after first removing moisture and debris is crushed in a mill for one hour. Product grinding quantitatively from the mill is served in an air classifier jet of dry air separation, particles more 45,0*10-6m in an amount of 5 kg is returned to the mill for regrinding. Formed in the amount of 245 kg per hour of fine fraction not larger than 45,0*10-6m served in the furnace, providing uniform heating of mineral raw materials at 200°C for 10 minutes, after which the product is cooled in the heat exchange device to the ambient temperature.

Example 1A

Example 1A is carried out in the conditions of example 1, but after the classifier are selected and served in a furnace product of grinding, the particle size of which is not more than 100*10-6m

Example 1B

Example 1B is carried out in the conditions of example 1, but after the classifier are selected and served in a furnace product of grinding, the particle size is less than 10*10-6m

When the heat treatment is appreciable agglomeration of the particles with the formation of large pieces of irregular shape increased the size of the A.

The sizes of the agglomerated particles less than 300*10-6m

Example 2

Example 2 is carried out in the conditions of example 1, but using schungite comprising mineral raw materials from the field Tekeli-koksuiski zone with a carbon content 3,998 wt. -%, silicon oxide 69,93% of the mass. and heat treatment of the separated fractions is conducted for 60 minutes at a temperature of 100°C.

Example 2A

Example 2A is carried out in the conditions of example 2, but after the classifier are selected and served in a furnace product of grinding, the particle size of which is not more than 100*10-6m

Example 3

Example 3 is carried out in the conditions of example 1, but using schungite comprising mineral raw materials of zazhoginsky deposits with a carbon content of 55.1% of the mass, silicon oxide 15.1% of the mass.

Example 3A

Example 3A is carried out in the conditions of example 3, but after the classifier are selected and served in a furnace product of grinding, the particle size of which is not more than 100*10-6m

Example 4

The use of enterosorbent for broiler chickens

The experience carried out on chickens-broilers cross "ISA" in cell method, poultry keeping. At day old form of control and experimental group.

Enterosorbent obtained in example 1, is introduced into the feed mixture from the experimental group by gradually mixing in bunkers in the amount of 0.1% of the ration or 1 g per 1 kg to the mA. The period of feeding and growth of broiler chickens is 42 days.

The control group fed food in the same amount and of the same composition, and that experienced, but without adding to the diet composition of enterosorbent.

To assess the health of the population the test and control groups consider the following indicators: weight - weighing calves and breeding average live weight, daily safety control; immunobiochemical the blood of chickens at slaughter.

The results of weighing indicators are shown in table 2.

Table 2
The dynamics of clinical and physiological indices
IndicatorsGroup Chicks
experiencedcontrol
Number of goals in the group4200042000
Live weight of chickens, g
- at the start of feeding (i.e. daily age)37,3±0,1237,3±0,13
after 7 days140±1,0123±1,0
- 14 days396±1,6376±1,6
- after 28 days1150±2,4998,5±2,2
- 42 days20001700
Safety, %99,297,3
Average daily gain, g46,739,5
The cost of feed for the rearing period on 1 kg gain, kg (conversion)1,581,70

As can be seen from the data presented, the live weight of the chickens in the experimental group compared to control after a week from the start of feeding, and continues to be great throughout the feeding period. Increased measure of security of broiler chickens (99,2% compared with the control 97,3%), and average daily increase in them (46.7 g for the experimental group, 39.5 g for control). This reduced the cost of feed for the rearing period on 1 kg gain (conversion) - 1,58 kg for the experimental group prot is at 1.70 kg for the control.

The results of biochemical analyses of the blood of broiler chickens at slaughter are shown in table 3.

Table 3
The biochemical composition of the blood of chickens-broilers-before slaughter
IndicatorsGroup Chicks
With the use of enterosorbentWithout the use of enterosorbent
Biochemical parameters
Total protein, g %4,8±0,34,3±0,25
Alkalinity, vol.%, CO45,6±0,0639,6±0,06
Calcium, mg %17,9±0,816,6±0,9
Phosphorus, mg %4,9±0,54,2±0,4
Indicators of natural resistance
Bactericidal activity of blood serum, %52,63±2,1347,80±2,14
The authors estimated the effect activity of blood serum, %18,71±0,5616,90±1,15
Phagocytic activity, %49,33±2,2546,14±2,27
Immunoglobulins%5,02±0,564,07±0,56

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

Thus, it is shown that the introduction of enterosorbent at the rate of 0.5 % per kg of feed as a Supplement to the diet of broiler chickens has had a tangible effect to stimulate growth energy, increased safety and a bird.

Example 5

The use of enterosorbent for laying hens.

The experience carried out at the cellular method, poultry cross "Hi-line". Form of control and experimental group.

Enterosorbent obtained in example 2 is introduced into the feed mixture from the experimental group by gradually mixing in bunkers in the amount of 0.5% of the diet. The observation period of laying hens is 150 days.

The control group fed food in the same amount and of the same composition, and experienced, names added to the diet of enterosorbent.

To assess the health of the population the test and control groups include the following criteria: egg production, preservation, and weight gain of laying hens, which are shown in table 4.

Table 4
Indices of egg production, preservation, and weight gain of laying hens when observed within 150 days
IndicatorsThe group of laying hens
experiencedcontrol
The number of goals2100021000
Egg production, %, eggs per day by the number of days
after 10 days0,79±0,070,78±0,07
- 25 days0,85±0,090,80±0,09

50 days0,92±0,050,82±0,05
- after 75 days0,95±0,040,85±0,04
The average weight gain of laying hens, g
before feeding12021185
after feeding (within 150 days)13051250
Safety for 150 days %98,3for 95.2

As can be seen from the presented data, the rate of egg production in the experimental group compared to control increased after 10 days added to the diet declared enterosorbent (0,79 versus 0.78), continued to exceed the control throughout the experiment and after 75 days reached a difference of 10%.

Increased rate of preservation of laying hens (98,3% compared with the control 95.2 percent)and the rate of egg production.

Example 6

The use of enterosorbent for pigs

The experience carried out on pigs of large white breed 100-110 days old. Enterosorbent obtained according to example 3 is introduced into the feed mixture from the experimental group by gradually mixing in bunkers in the amount of 0.2% of the diet. The control group fed food in the same amount and of the same composition, and that experienced, but without adding to the ration of enterosorbent.

Indicators of weight gain are shown in t the blitz 6.

Table 6
The growth in weight of pigs
IndicatorsThe control groupExperienced group
The number of animals in the group, goals3030
The average weight before the beginning of the input, kg31,3±0,632,5±0,2
The multiplicity of input per day3 times3 times
The average weight in 30 days after application, kg44,2±0,347,6±0,28
Average daily gain, kg0,4320,503

As can be seen from the data presented in the beginning of the trials the average weight of the experimental and control groups were virtually identical, and after 30 days of application of the enteric average weight in the experimental group compared with the control 3.4 kg (47,6±0.28 kg for the experimental group, 44,2±0.3 kg for the control), average daily gain for itoi group 71 g more than in control (0,503 and 0,432 kg, respectively).

Example 7

Research on the efficiency of enterosorbent for the treatment of diarrhoea of calves

The experience carried out on calves at the age of 3-12 days.

Sick animals on the principle analogues are divided into 2 groups of 20 calves each. Calves from the experimental group for the treatment of diarrhea apply enterosorbent obtained according to example 2, the rate of 1.0% of the mass. on the weight of the feed, the control group, the drug was not given, only the feed.

Animals as experimental and control groups are daily observation, taking into account the clinical condition, appetite, recovery time. Cured believe animals have within 10 days from the start of treatment disappeared clinical signs of disease, and there was no relapse. The results are shown in table 7.

Table 7
The efficiency of enterosorbent for the treatment of diarrhoea of calves
IndicatorsWithout enterosorbentWith enterosorbent
The number of animals in group2020
Multiplicity of day (time) 22
Time (days)-7
Recovery period (days)-3
Recovered (goal.)1719
Therapeutic efficacy, %85%95%

It is established that the application of enterosorbent in the experimental group led to improvement of the General condition of the animals with subsequent recovery at 2-3 days of treatment. Therapeutic efficacy of enterosorbent was 95% in comparison with the control, where the drug did not ask.

Example 8

Determine the effectiveness of the claimed enterosorbent as adsorbent of mycotoxins from the body of the bird

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

Studies conducted on chickens-broilers cross "ISA". In days age form the control and experimental groups 42000 goals each.

In broilers ' diet of the experimental group injected with daily age-related disease is enterosorbent, obtained in example 1, in an amount of 0.1% per kg of feed as a Supplement to the diet. Enterosorbent is introduced into the feed mixture in the feed of poultry farms by gradually mixing in bunkers.

To monitor the dynamics of changes in the content of mycotoxins in the tissues of birds (liver, stomach and its contents, and feces), check-in and at the bottom of the control and experimental groups selected samples by irrevocable removal of 20 birds from each group. Determination of mycotoxins comply with the normative-technical documentation (for aflatoxin B1 - GOST 30711-2001, T2-toxin - guidelines for the determination of T-2 toxin in biological tissues and extracts (Approved December 27, 1989) (B. I. Antonov, Yakovlev TF, Deryabina VI Laboratory tests in veterinary medicine: biochemical and mycological. - M.: Agropromizdat, 1991, str)for zearalenone f-2 Method of determining mycotoxin f-2 (zearalenone) in organs and tissues of animals (Approved may 31, 1987) (B. I. Antonov, Yakovlev TF, Deryabina VI Laboratory tests in veterinary medicine: biochemical and mycological. - M.: Agropromizdat, 1991, p.151), ochratoxins - Methods of determination of ochratoxin a in organs and tissues of animals (Approved April 10, 1984) (B. I. Antonov, Yakovlev TF, Deryabina VI Laboratory studies in ve is urinaria: biochemical and mycological. - M.: Agropromizdat, 1991, p.153).

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

The contents of the various mycotoxins in samples taken from the control and experimental groups at the check in, it was about the same, this can be explained by the fact that birds of both groups were taken from the same brood; feed at the beginning of the experiments were identical.

The data of table 8 show that the content of mycotoxins in the internal organs of the control group of chickens was significantly higher compared with the experimental group receiving the enterosorbent as a feed additive. The growth of mycotoxins in the contents of the stomach and faeces from the experimental group shows that the mineral raw material binds toxins in the stomach and easily removes them from the body of birds. The results clearly indicate that the introduction of enterosorbent in the feed of broiler chickens has had a tangible effect in the removal of mycotoxins from the body.

Example 9

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

The experience carried out on chickens-broilers cross "ISA". In days age on the principle of steam-analogues form of control and experimental group (n=40).

Both groups experience the t of the seed sodium nitrate (NaNO 3at the rate of 50 mg NaNO3per kg of feed three times during the life of birds.

The experimental group, since the check is fed announced enterosorbent obtained according to example 2, which is added to the feed mixture by the method step of mixing at the rate of 0.5% per kg of feed.

At the bottom (after 42 days) take samples of liver and muscle tissue. Methods of sampling and preparing them for the test comply with the normative and technical documentation (GOST 9792-73), they are performed in accordance with techniques passed state certification GOST 29300-92 (ISO 3091-75) for nitrate, GOST 29299-92 (ISO 2918-75) for nitrite.

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

Table 9
The content of nitrates and nitrites in organs of broiler chickens
№ p/pSample description bodyNitritesNitrates
The control group (average results obtained with 40 goals), mg/kg
1Muscle tissue7±0,34 Not found
2Liver22,04±0,47Not found
Experimental group (average results obtained with 40 goals) mg/kg
1Muscle tissue3,56±0,56***Not found
2Liver5,32±0,17***Not found
Note: Data is reliable compared with the control group: * P≤0,05, ** - P≤0,01, *** P≤0,001

The data of table 9 show that the content of nitrates and nitrites in the experimental group was significantly less than in the control, the introduction of enterosorbent in the diet of broiler chickens by reducing the concentration in muscle tissue: nitrites almost 2-fold in the liver by 4 times, the content of nitrates in both groups were not found. This is because in the body of birds nitrates unstable and quickly moving into nitrites.

Example 10

Determine the effectiveness of the claimed enterosorbent as the sorbent is of heavy metals from the body of the bird

The experience carried out on chickens-broilers cross "ISA". In days age form the control and experimental groups of 20 goals each.

Both groups are subjected to the simultaneous priming 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. Feed pre-treated with 10%-governmental solutions of acetates data of heavy metals, prepared with distilled water. Thus, the seed is produced every time a set of feed.

Experimental group, starting with the settlement receives enterosorbent obtained according to example 2, the rate of 0.2% per kg of feed as a Supplement to the diet.

After slaughtering (after 42 days) take samples of muscle and bone tissue and determine the content of lead and cadmium. Methods of sampling and preparing them for the test corresponds to the normative-technical documentation (GOST 26929-94); they are conducted according to guidance by the atomic absorption method for determination of toxic elements in food 01-19 (47-11-92). The analysis of the objects of study on the content of heavy metals conduct methods, certified metrological service of Gosstandart of the Russian Federation: for lead (GOST 26932-86), cadmium (GOST 26933-86). Studies of organs and tissues of birds on the content of heavy metals is performed on an atomic absorption spectrometer (GOST 30178-96). Received financial p the tats are presented in table 10.

Table 10
The content of heavy metals in muscle and bone tissues of broiler chickens of the control and experimental groups
GroupIndex
White muscleRed musclesBone
Cd (µg/g)Pb (µg/g)Cd (µg/g)Pb (µg/g)Cd (µg/g)Pb(µg/g)
control0,252±0,0550,481±0,1210,264±0,0540,712±0,1411,247±0,4115,523±,954
experienced0,004±0,06**0,125±of 0.081**0,006±0,040**0,179±0,156**0,087±0,043**0,265±0,139
Note: Data is reliable compared to the control group: * - P≤0,05 ** P≤0,01 *** P≤0,001

The data of table 10 show that the content of cadmium and lead in the experimental group was significantly less than in the control, the introduction in the diet of broiler chickens offered enterosorbent reduces the content of heavy metals:

in white muscle: cadmium in 82.6 times, lead - 3.7 times;

in red muscle: cadmium 4 times, lead - 4.9 times;

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

Example 11

Determination of antioxidant properties of the claimed enterosorbent

Antioxidant properties of the claimed enterosorbent investigate its influence on the processes of lipid peroxidation. The degree of intensity of the process of lipid peroxidation (LPO) is judged by the accumulation of secondary LPO products - malondialdehyde (MDA) in the reaction with 2-thiobarbituric acid [Goncharenko, M.S., Method of assessing lipid peroxidation / Mashonkina, Amolatina // Laboratory work. - 1985. No. 1. - p.60-61].

Experiments carried out on white rats. Before conducting experiments in animals kept on a two-week quarantine. Experimental in the morning once give complete feed from the nursery of the Federal state institution Federal center for Toxicological and radiation safety of animals (Kazan). Water set in descending order. Experimental and control gr is PI animals formed on the principle of analogues. Throughout the experience the animals were housed under identical conditions and feeding.

For experimental studies using T-2 toxin as one of the most common and the most dangerous mycotoxins. To evaluate the effectiveness of the claimed enterosorbent used as the comparison drug enterosorbent "Polisorb" production "POLISORB, Russia.

The experience of Provat 50 males nonlinear white rats, animals are divided into 5 groups of 10 pieces:

group 1 - biological control, during the whole period of experiment received a "clean" feed that does not contain mycotoxins;

group 2 - animals during the entire period of experiment received a "clean" feed that does not contain mycotoxins, mixed with the claimed enterosorbent in an amount of 5 g/kg feed;

group 3 - animals asked "toxic" food contaminated with the mycotoxin T-2 (1.0 mg/kg feed);

group 4 animals received "toxic" food contaminated with the mycotoxin T-2 (1.0 mg/kg feed), mixed with enterosorbent "Polisorb" in the amount of 5 g/kg feed;

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

The inventive sorbent was taken at the maximum dose (5 kg/t of feed, according to instrukciya use). In order to dose the introduction of the same in the same amount was taken to compare the sorbent Polisorb".

The duration of the experiment 10 days.

The consumption of rat feed and water all of the groups did not differ. Survival in all groups was 100%.

On the ninth day take blood for research by decapitate ten rats from each group. The degree of intensity of the process of lipid peroxidation (LPO) is judged by the accumulation of secondary LPO products - malondialdehyde (MDA) in the reaction with 2-thiobarbituric acid (TBA).

To study selected 1.5 ml of blood with heparin. By centrifugation (5 min, 3000 rpm) to separate the plasma from the formed elements of blood. Next, analyze the content of TBA-active products separately in plasma and in erythrocytes. For this RBC mass is washed twice with 10 ml isotonic. After washing the red blood cells are precipitated by centrifugation for 10 min at 3000 rpm, Washed erythrocytes hemolyticus in distilled water at a ratio of 1:5. For research selected 0.3 ml plasma and 0.3 ml of hemolysate, to which was added 2.4 ml H2SO4and 0.3 ml of 10% phosphate-tungsten acid, mix thoroughly. After 10 minutes the resulting precipitate was separated by centrifugation about 3000 rpm, the Supernatant discarded, and obtained the precipitate is washed twice with 1 ml of water, dissolved in 3 ml of water, add 1 ml of freshly prepared aqueous solution thiobarbiturate acid in acetic acid (80 mg thiobarbiturate acid are dissolved by heating in 5 ml of H2O and 5 ml glacial acetic acid). The color reaction is performed in a closed chemical test tubes at 96°C. the Reaction is stopped after 60 min, cooling the sample in cold water. For removal of turbidity before spyektrofotomyetrirovaniya samples centrifuged for 15 minutes at 3000 Rev/min the Optical density measured on the spectrometer SF-26 at a wavelength of 532 nm in cuvettes with optical path length of 10 mm as a control using distilled water. Calculate the amount of malondialdehyde (MDA) is carried out with regard to cultivation by the formula C=E/έ where E is the optical density of the sample, έ - molar ratio instinctive equal to 1.56×10-5mol-1/cm-1With the concentration of MDA was expressed as mcmash 1 ml of plasma and 1 ml of erythrocyte mass. The values of MDA in plasma and erythrocytes summarize.

The results are shown in table 11.

Table 11
The content of malondialdehyde (×10-5mol-1cm-1in the blood of rats of groups 1-5 in 10 days
the group Index
10,784
20,724
30,971
40,892
50,823

Register a natural increase in the content of malondialdehyde (MDA) in the blood of rats 3-5 groups treated with "toxic" food, compared with groups 1 and 2 of the control.

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

The content of malondialdehyde in blood of rats 4 and 5 groups are pretty close, although in 5 group this value is slightly lower (0,823×10-5mol-1cm-1against 0,892×10-5mol-1cm-1for the treated group "Polisorb"), this means that the claimed enterosorbent more actively linked peroxide products of metabolism than Polisorb".

In the study of adsorption properties of the claimed enterosorbent on from the oseney to various kinds of microorganisms (Escherichia coli, Salmonella typhimurium, Staphylococcus aureus, Clostridium perfringes) it was found that they were significant adsorption 63,2-91,8%, which is comparable to, and sometimes far exceeds the adsorption properties of the prototype.

Example 12

The adsorption action of the claimed enterosorbent against microorganisms shown in the example of the culture of Escherichia coli.

1 day before the start of the experiment on the beveled mastopathy agar (MPA) conduct culture of Escherichia coli. The crops are placed in the incubator for 24 hours at 37°C. after 24 hours, spend aspirates cultures with sterile distilled water, filtered through a sterile filter and prepare a working suspension of microbes, by standardizing on the standard optical density 2 units Simultaneously prepare a working suspension with the claimed enterosorbent obtained in example 1. To do this in a sterile tube filled with sterile isotonic solution of 4.5 ml and add 0.5 ml of the working suspension culture and 25 mg of sterile enterosorbent (the number used to experience enterosorbent due to maximum recommended dose of the enteric - 5 g/kg feed; to maintain a specified ratio calculation was 5 g/l of sterile isotonic). Working suspension with the claimed enterosorbent placed in the apparatus for shaking the samples for 1.5 hours. Next, perform a serial spy who s this mixture 1:10, 1:100, 1:1000, 1:10000, of which conduct crops on MPA in Petri dishes (three sowing at each dilution) from the tube by means of "lawn" to 0.1 ml of the resulting dilutions. Cups were placed in the incubator for 24 hours at 37°C.

In parallel, set the controls, i.e. perform the same steps for:

- tubes with a working suspension of microbes without the claimed enterosorbent (main control, compared to which shall count adsorption);

- tubes with isotonic and claimed enterosorbent without microbes (control for sterility, to verify the validity of the results that with water or declare enterosorbent have not made other microbes);

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

After 24 hours, visually shall count the number of grown colonies in three cups, to count SOME took the results of the largest breeding with the least number of grown colonies, and take the average result - this figure was then multiplied by the degree of dilution, the resulting figure was compared with the control culture and the difference of numbers considered as adsorption and were calculated in percentages. Studies were performed 3-fold with stat. processing.

After 24 hours visually shall count the number of Viro the Shih colonies in three cups, to calculate KOE took the results of the largest breeding with the least number of grown colonies (1:10000), take the average result, which is multiplied by the degree of dilution (105), and the resulting figure is compared with the control crops.

The difference is considered as adsorption and converted to percentages. Research carried out three times using statistical processing.

The results are presented in table 12.

Table 12
The adsorption action of the claimed enterosorbent in relation to the culture of Escherichia coli, CFU/ml (×105)
CFU/mlThe rate of adsorption activity, %
Control50
Polisorb (prototype)1766
Invention1570

According to table 12 shows that the adsorption against Escherichia coli declared enterosorbent amounted to 70%, and the Polisorb - 66%, i.e. declared enterosorbent shows better adsorption properties is against pathogenic strain of Escherichia coli in comparison with the prototype.

Example 13

Testing chelators carried out with the use of products obtained according to examples 1A, 1B, 2A, 3A, 4A, similarly to the description provided in example 12.

The results are given in table 13.

Table 13
The adsorption action of the claimed enterosorbent in relation to the culture of Escherichia coli, CFU/ml (×105)
CFU/mlThe rate of adsorption activity, %
Control50
Polisorb (prototype)1766
Example 11570
Example 1A2158
Example 1B1962
Example 2A1962
Example 3A2060

According to table 1 it is evident, the adsorption against Escherichia coli by chelators obtained in examples 1A, 1B, 2A, 3A amounted to smaller values. From the presented data it follows that the particle size is a significant factor in determining the adsorption properties against pathogenic strains of Escherichia coli, and exhibits the best properties in the case when the maximum particle size is in the range of up to 45*10-6m and the average median size 15*10-6m

Example 14

Determine the effectiveness of the claimed enterosorbent as adsorbent of mycotoxins from the body of the bird

Tests of enterosorbents in this example was conducted with the application of enterosorbents, obtained according to examples 1A, 1B, similarly to the description in example 8. Analogously to example 8 in the diet of broilers experimental group injected with daily age to slaughter enterosorbent obtained in example 1, in an amount of 0.1% per kg of feed as a Supplement to the diet. The efficiency of adsorption was estimated by the content of one of the mycotoxins - Aflatoxin B1 was tested using the sorbents obtained according to the procedure described in examples 1, 1A and 1B. The results are shown in table 14.

Thus, the claimed polyfunctional enterosorbent on the basis of schungite comprising mineral raw materials, with the high sorption properties against a broad range of toxic substances, including mycotoxins, nitrates, nitrites, heavy metals, and also shows the properties of antibacterial and antioxidant action.

The inclusion of the enterosorbent in the diet of farm animals and birds have a positive effect on increasing gain, egg production, preservation, resistance. The obtained results give reason to recommend claimed enterosorbent for the prevention and treatment of poisoning and diseases of the gastrointestinal tract of various etiologies in farm animals and birds.

Polyfunctional enterosorbent on the basis of natural siliceous minerals, characterized in that as a natural siliceous mineral raw materials it contains schungite comprising mineral raw materials with a silicon dioxide content of 15.0-70,0 wt.%, and the average median particle size of enterosorbent is 15,0·10-6m



 

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1 ex

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40 cl, 4 dwg, 24 tbl, 144 ex

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20 cl, 2 dwg, 13 tbl, 22 ex

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6 tbl, 7 ex

Radioprotector // 2428192

FIELD: medicine.

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6 tbl, 7 ex

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