RU2477614C1 - Method for production of complex biologically active fodder additive for farm animals, birds and fish with probiotics and medicinal herbs - Google Patents

Abstract

FIELD: food industry.

SUBSTANCE: invention relates to fodder production, in particular, to a method for preparation of a biologically active fodder additive for farm industry animals, birds and fish. One performs separate in-depth cultivation of Bacillus subtilis VKPM V-8130, Bacillus subtilis VKPM V-2984, Bacillus subtilis VKPM V-4099 and Bacillus licheniformis VKPM V-4162 strains to produce liquid cultures. One performs solid phase fermentation of Bacillus subtilis VKPM V-8130, Bacillus subtilis VKPM V-2984 and Bacillus subtilis VKPM V-4099 liquid cultures. One mixes the cultures at a ratio of 6:6:1, accordingly, to produce 65 l of the mixture which is applied on sterile beet pulp (in an amount of 200 kg) treated with a cellulolytic enzyme and enriched with Saccharomyces cerevisiae fodder yeasts fermentolysate, such pulp representing a preliminarily prepared carrier for solid phase fermentation implementation. To produce the Saccharomyces cerevisiae fodder yeasts fermentolysate one prepares the following components mixture: molasses- 255-265 g, disubstituted potassium phosphate -98-102 g, magnesium sulphate - 25-26 g, Saccharomyces cerevisiae feed yeast - 3 kg and water -up to 30 l. The mixture is sterilised at a temperature of 120°C during 30 minutes and added to the carrier wherein one introduces 4 l of a solution of Celloviridin or Cellulox F containing cellulase in an amount of no less than 2000 units/g of the carrier; pH is conditioned to 6.0-6.5; the mixture is thoroughly stirred, maintained during 2 hours at a temperature of 45-50°C. Solid phase fermentation is performed under restricted oxygen access conditions at a temperature of 45-50°C, pH being 7.5-8.0 and batch moisture content equal to 43-48%, during 48-50 hours. 65 l of Bacillus Hcheniformis VKPM V -4162 liquid culture containing no less than 5.6x10⁸ CFU/g are added into the manufactured product. The mixture is thoroughly stirred and dried till moisture content is equal to 8-10%; then one adds dry powders of purple echinacea herb and Saint-Mary-thistle fruits in an amount of 20-50 g of the echinacea powder and 20-50 g of holy thistle powder per 1 kg of the end product. The produced mixture is stirred during 0.5 hour and crushed till a homogeneous mass production.

EFFECT: invention ensures animals, birds and fish productivity enhancement, livestock liveability enhancement, intensification of metabolic processes in the organism and animal mycotoxication prevention.

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Description

The invention relates to feed production, in particular to methods for the preparation of biologically active feed additives for animals, poultry, fish, which increase the efficiency of digestion, have a versatile effect on the animal organism, and allow their use both for the treatment and prevention of gastrointestinal diseases, and for stimulation growth and productivity.

Probiotics - live microbial feed additives that have a beneficial effect on the animal's body by improving the intestinal microbial balance - are an important product in the world market. They positively affect the host’s body, contribute to the restoration of digestion, biological status, immune response, and increase the effectiveness of vaccinations. The use of probiotics significantly reduces the cost of treating diseases in animals, increases the productivity of the latter and improves product quality, which ultimately increases the profitability of livestock production.

To obtain new biologically active drugs, complexes of probiotics with prebiotic substances are combined. The progressive forms of new generation drugs include sorbed forms of probiotics. Sorbed probiotics contain bacteria immobilized on solid sorbent particles. Due to chemical and electrostatic forces, the interaction of such forms with the intestinal wall is higher. Sorbent accelerates detoxification and reparative process. As a sorbent, a prebiotic can be used.

Prebiotics are a class of drugs for the regulation of intestinal microflora, which is becoming increasingly popular. Prebiotics - substrates that stimulate the natural microflora, are not digested and are not absorbed in the stomach and small intestine. Once in the large intestine, prebiotics are used as a nutrient medium for normal microbiota. In mammals, in the first days after birth, the main prebiotic substrate is lactulose, which is part of milk. With the transition to mixed feeding of the substrate, contributing to the growth of normal microbiota, the structural elements of plant tissues, various polysaccharides (pectins, inulin, etc.) become. Dietary fiber also has other important functions: normalize motility, prevent constipation, and absorb toxins.

Echinacea has immunomodulatory properties. Chemical composition: all plant organs contain polysaccharides, essential oil (flowers up to 0.5%, grass up to 0.35%, roots up to 0.25%). The main component of the essential oil is non-cyclic sesquiterpenes. Echinacoside glycoside, betaine (0.1%), resins (about 2%), organic acids (palmitic, linoleic, cerotinic) and phytosterols were found in the roots. The main active substances with immunostimulating activity are polysaccharides. Echinacea preparations have a therapeutic effect in various pathological conditions by increasing the body's natural defenses. The stimulating effect of echinacea on the immune system of adults, young animals with an unstable immune system, and old individuals with reduced functions of this system is proved. Echinacea preparations are used for diseases associated with a weakening of the functional state of the immune system caused by chronic inflammatory diseases, exposure to chemotherapeutic drugs, and prolonged antibiotic therapy. When taking Echinacea preparations in cases of metabolic disorders (for example, liver disease), exposure to various chemical compounds of a toxic nature contained in feed (heavy metals, pesticides, insecticides, fungicides), a stimulation of the immune system is observed. Echinacea preparations also exhibit antibacterial, antiviral and antimycotic properties. Echinacea drugs inhibit the growth and reproduction of streptococcus, staphylococcus, E. coli, influenza viruses, are effective in inflammatory diseases (including gynecological disorders), diseases of the upper respiratory tract, in various wound processes (trophic ulcers, osteomyelitis). Echinacea purpurea polysaccharides increase the resistance of immunodeficient mice to infections of Candida albicans and Listeria monocytogenes. Echinacea purpurea polysaccharides also stimulate macrophage cytotoxicity against tumor and microbial cells.

Milk thistle is used to prevent various lesions and the liver. Milk thistle is unique in that it helps cleanse and form new liver cells. Milk thistle preparations increase the protective properties of the liver to infection and poisoning, stimulate the formation and excretion of bile. The positive effect of the plant affects both the liver and the entire gastrointestinal tract. It is advisable to use milk thistle in the form of a powder, since it works at the micro level, cleansing liver cells.

The main active ingredients of milk thistle are flavonoids and flavonolignans (silibin, silikristin, silidianin). In addition, it contains alkaloids, saponins, fatty oil (up to 25%), proteins, vitamin K, resins, mucus, tyramine, histamine, as well as macro and micronutrients. The healing properties of leaves, roots, and most importantly - milk thistle seeds. The seeds contain fatty and essential oils, vitamin K, resins, mucus, tyramine, flavonoids, as well as macro and micronutrients. Milk thistle goes well with various herbs, herbs and tinctures for the treatment of certain diseases. Milk thistle - a plant containing essential oils and flavonoids (silybin, silicocristine, silyidiamine), which have a powerful detoxifying, hepatoprotective, antioxidant effect, is indispensable in restoring human microflora. 2 preparations were obtained from milk thistle: “Mariol” in the form of a powder with the properties of a flavonoid (silymarin), produced according to the Republic of Uzbekistan Ministry of Health of the Russian Federation 000554.Р.643.11.98; and "Mariol-MK" in the form of milk thistle, calendula and chamomile oil, produced in accordance with RU Ministry of Health of the Russian Federation 001625.P.643.06.2000. The main active ingredients of the oil are essential (essential) polyunsaturated acids (at least 55 g / 100 ml), biogenic amines, phospholipids, tocopherols, carotenoids, essential oils; Vitamins A, D, E, K, F. RU 22171562 C2, 11.27.2003.

It is known to use plant extracts of spotted milk thistle (Silybum marianum (L.) Gaertn) and the extract of the roots and grass of Echinacea purpurea (Echinacea purpurea L.) as part of a biologically active agent encapsulated in phospholipids intended for the prevention of colibacteriosis in birds. The agent in the form of a water-soluble form of application is added to the feed, which improves the resistance of the poultry to infectious diseases. RU 2355410 C2, 05.20.2008.

A known method of producing a biologically active feed additive, comprising two stages of its production: deep aerobic cultivation of a probiotic cellulolytic bacillus - strain Bacillus subtilis VKPM B-8130, which is part of the drug Probitsel - additives to animal feed, obtaining a liquid culture (first stage), and subsequent solid phase fermentation of plant materials with difficult access of oxygen (second stage). The process is carried out in the presence of sodium bicarbonate (NaHCO ₃ ) to bind acetyl-CoA and neutralize acidic fermentation products, thereby using the Neuberg metabolic trap method for yeast fermentation. As plant raw materials, sea buckthorn, dogrose, sunflower shrimp or cake, waste from flour milling, cereals production are used, RU 2346463 C2, 02.20.2009.

The closest analogue of the claimed method is a method for producing a probiotic feed additive of Farms KM, which is obtained by separate deep cultivation of strains of Bacillus subtilis VKPM B-8130, Bacillus subtilis VKPM B-2984, Bacillus subtilis VKPM B-4099 and Bacillus licheniformis VKPM B-4162 and subsequent solid phase fermentation. Why the obtained liquid cultures of Bacillus subtilis VKPM B-8130, Bacillus subtilis VKPM B-2984 and Bacillus VKPM B-4099 are mixed in a ratio of 6: 6: 1 to a volume of 65 l, applied to 200 kg of sterile beet pulp, which is pretreated with cellulolytic enzyme dissolved in a nutrient medium containing 30 l of water: molasses - 255-265 g, potassium disubstituted phosphate - 98-102 g, magnesium sulfate - 25-26 g, yeast - 1.8-2.2 g, the pH is adjusted to 6.0-6.5 and incubated for 2 hours at a temperature of 45-50 ° C. After pretreatment of beet pulp, a mixture of culture liquids Bacillus subtilis VKPM B-8130, Bacillus subtilis VKPM B-2984 of 30 l per load and Bacillus subtilis VKPM B-4099 - 5 l per load are added to it, the pH is adjusted to 7.5-8.0 and humidity kneading up to 43-48%. The mixture is thoroughly mixed, packaged in bags and sent to the fermentation chamber for solid phase fermentation under conditions of limited oxygen access at a temperature of 45-50 ° C for 48-50 hours. After completion of the solid phase fermentation process, 65 L of Bacillus licheniformis liquid culture are added to the resulting mass. VKPM B-4162, containing at least 5.6 × 10 ⁸ CFU / g, the mixture is thoroughly mixed and dried to a moisture content of 8-10%. The resulting product is transferred to a two-stage crushing to obtain a homogeneous mass with a particle size corresponding to the requirements of the feed industry products. RU 2412612 C1, 02.27.2011.

In the present invention, the method of producing a complex biologically active feed additive for farm animals, poultry and fish with probiotics and medicinal herbs is characterized by the fact that separate deep cultivation of strains of Bacillus subtilis VKPM B-8130, Bacillus subtilis VKPM B-2984, Bacillus subtilis VKPM B- 4099 and Bacillus licheniformis VKPM B-4162, obtaining their liquid cultures and subsequent solid-phase fermentation, for which the obtained liquid cultures of Bacillus subtilis VKPM B-8130, Bacillus subtilis VKPM B-2984 and Bacillus subtilis VKPM B-4099 are mixed in a ratio of 6: 6: one, respectively, to obtain 65 l and applied to a pre-prepared carrier for carrying out solid-phase fermentation - sterile beet pulp in an amount of 200 kg, treated with a cellulolytic enzyme and enriched with fermentolizate of Saccharomyces cerevisiae fodder yeast, for which preparation: molasses - 255-265 g, potassium phosphate disubstituted - 98-102 g, magnesium sulfate - 25-26 g, fodder yeast Saccharomyces cerevisiae - 3 kg and water up to 30 l, the mixture is sterilized at a temperature of 120 ° C for 30 min and added to a carrier in which 4 l is introduced a solution of celloviridine or cellulox F with a cellulase content of at least 2000 u / g of the carrier, the pH is adjusted to 6.0-6.5, the mixture is thoroughly mixed, incubated for 2 hours at a temperature of 45-50 ° C. Solid phase fermentation is carried out under conditions of limited oxygen access at a temperature of 45-50 ° C, pH 7.5-8.0 and a kneading humidity of 43-48% for 48-50 hours, 65 L of Bacillus licheniformis VKPM B liquid culture are added to the resulting product -4162, containing not less than 5.6 × 10 ⁸ CFU / g, the mixture is thoroughly mixed and dried to a moisture content of 8-10%, after which dry powders of Echinacea purpurea and Milk Thistle are added at the rate of 20-50 g of Echinacea powder and 20 -50 g of milk thistle powder per 1 kg of the final product, the resulting mixture is stirred for 0.5 h and exposed crushing until a homogeneous mass.

The technical result of the invention is to increase the manifestation of prophylactic properties and increase the spectrum of biological action due to the content in the feed additive of crushed powder of medicinal plants - Echinacea purpurea grass and spotted milk thistle. Along with this, an increase in the productivity of farm animals, poultry, and fish, an increase in the safety of the livestock, an increase in metabolic processes in the body, and prevention of mycotoxication of animals are provided.

A significant distinctive feature of the claimed method is not the fact of using probiotic cultures of strains of Bacillus subtilis VKPM B-8130, Bacillus subtilis VKPM B-2984, Bacillus subtilis VKPM B-4099 and Bacillus licheniformis VKPM B-4162, which are part of the probiotic feed additive Farm KM, a enrichment of the solid-phase fermentation carrier with Saccharomyces cerevisiae feed yeast fermentolizate, prepared in the presence of cellulolytic enzymes and mineral salts, which provides an additional deeper cleavage of the cell wall of the nucleus ments. At the final stage of mixing probiotic components, dry powders of Echinacea purpurea and Milk Thistle are added. The resulting biologically active feed additive is named ProStor.

The ProStor preparation contains a set of the most important enzymes: cellulase, endoglucanase, amylase, protease, lipase, organic acids, biologically active substances, vitamins and amino acids, immunoactive peptides - products of probiotic metabolism. The preparation includes specially treated beet pulp containing bactericidal phytocomponents of beet pulp, terpenes and terpenoids, phytoparticles, microsorbents, and prebiotics - beet pectins. The composition is enriched with Saccharomyces cerevisiae fodder yeast obtained by special technology. The combination of mannan oligosaccharides and beta-glucans entering the cell walls of yeast serves as an additional prebiotic. The phytocomponents of echinacea and milk thistle expand the spectrum of the biological effect of the drug.

The invention is illustrated by the following example.

Example 1. A separate deep cultivation of strains of Bacillus subtilis VKPM B-8130 and Bacillus subtilis VKPM B -2984 is carried out on a nutrient medium containing, g / l:

molasses 24-26 corn gluten 14-16 corn extract 19-21 disubstituted potassium phosphate 1-3 sodium chloride 9-11 magnesium sulfate 0.4-0.6 water rest.

The cultivation of the strain of Bacillus subtilis VKPM B-4099 is carried out on a nutrient medium containing, g / l:

molasses 14-16 corn gluten 9-11 corn extract 9-11 disubstituted ammonium phosphate 1-3 sodium chloride 9-12 magnesium sulfate 0.4-0.6 water rest,

and strain Bacillus licheniformis VKPM B-4162 - on a nutrient medium containing, g / l:

molasses 24-26 corn extract 18-22 disubstituted potassium phosphate 1-3 sodium chloride 9-11 magnesium sulfate 0.4-0.6 water rest,

within 18-24 hours

The obtained liquid cultures of Bacillus subtilis VKPM B-8130, Bacillus subtilis VKPM B-2984 and Bacillus subtilis VKPM B-4099 are mixed in a ratio of 6: 6: 1, respectively, to obtain a final volume of 65 l.

As a carrier for solid phase fermentation, 200 kg of beet pulp is used, which is moistened with 10 l of water. Sterilization of the carrier is carried out with constant stirring and a temperature of 104-110 ° C for 30 minutes, for which sharp steam is supplied to the mixer from the steam generator until the sterilization temperature is reached. The temperature of the coolant is maintained at 120 ° C; at the end of the sterilization process, the medium is cooled to 48-50 ° C. After that, beet pulp is treated with a cellulolytic enzyme and simultaneously enriched with yeast fermentolizate, for which 30 l of a suspension of thermally inactivated fodder yeast Saccharomyces cerevisiae in a molasses solution with mineral salts is prepared as follows:

molasses is mixed - 260 g (1.3 g per 1 kg of carrier), K ₂ HPO ₄ - 100 g (0.5 g per 1 kg of carrier), M _g SO ₄ - 26 g (0.13 g per 1 kg of carrier ), Saccharomyces cerevisiae fodder yeast - 3.0 kg and water up to 30 L, are sterilized in a fermenter at a temperature of 120 ° C for 30 minutes and introduced into a mixer with a cooled sterile carrier. In 4 l of water, dissolve celloviridin or cellulox F with a cellulase content of at least 2000 u / g based on 200 g per total volume of liquid, and add it to the mixer. The mixture is thoroughly mixed, the pH of the mixture is adjusted to 6.0, and maintained at a temperature of 45-50 ° C for 2 hours.

After the process of fermentolysis of the resulting mixture, culture liquids Bacillus subtilis VKPM B-8130, Bacillus subtilis VKPM B-2984 of 30 l per load and Bacillus subtilis VKPM B-4099 - 5 l per load are added, the pH is adjusted to 8.0 and batch humidity up to 45%. The mixture is thoroughly mixed, packaged in bags and sent to the fermentation chamber for solid-phase fermentation at a temperature of 45 ° C for 48 hours. After completion of the solid-phase fermentation process, 65 l of Bacillus licheniformis VKPM B-4162 liquid culture containing at least 5.6 are added to the resulting mixture. × l0 ⁸ CFU / g, the mixture is thoroughly mixed and dried to a moisture content of 9%. Dry powders of Echinacea purpurea herb and Milk Thistle fruit are added to the resulting product, based on 25.0 g of Echinacea powder and 50.0 g of Milk Thistle powder per 1 kg of the final mixture. The dried products are mixed for 0.5 h, transferred to a two-stage crushing to obtain a homogeneous product with a particle size that meets the requirements of the feed industry products.

The obtained ProStor feed additive is used to increase the digestion efficiency and prevent mycotoxification of farm animals and poultry, by introducing it into feed diets at a dose of 1 kg / t by direct input into compound feed or by introducing it into premixes, BMVD, superconcentrates or other components of compound feed.

Indicators of body weight of mice on different diets are presented in table 1.

Table 1

Indicator Experience, diet with ProStor (yeast fermentolizate, 5% milk thistle, 2.5% echinacea) Control 2, diet with the drug Farm KM Control 1, diet without feed additives Total body weight at the beginning of the experiment, g 113.2 ± 9.4 131.5 ± 12.1 122.5 ± 10.5 Total body weight at the end of the experiment, g 134.3 ± 10.8 136.6 ± 12.1 125.6 ± 10.4 Weight gain, g 21.1 5.1 3,1 Weight gain,% 680.6 164.5 100.0

The ProStor test performed on mice showed that ProStor is more effective than KM Farms in the diet of experimental animals.

The physiology and rations of farm animals are taken into account in the ProStor dietary supplement formulation.

Application for broilers - daily starting from the 5th day and the entire growing period with compound feed, which includes 1 kg / t ProStor;

application for highly productive dairy cows - for periods of a month before and after calving, at the rate of 1.5 -2.0 kg / t ProStor.

When fattening piglets on a diet with ProStore feed additive, it was shown that the animals of the experimental group exceeded the corresponding indicator of the control animals by 5.8% in terms of gain in live weight, which confirms the biological effectiveness of the drug.

ProStor was tested on laying hens. Feeding laying hens with compound feed with the introduction of 1 kg / t of the feed additive ProStor for 6 months allowed us to increase egg production by 21%, reduce fetal mortality by 4%. The results confirm the effectiveness of the drug when using feeds contaminated with mycotoxins due to the physical structure of the phytosorbent and the total charge of pectin substances, which are a multicharged structure and have a high sorption ability. The feed additive also has protective properties, which is especially important when the feed is infected with non-polar compounds that adversely affect the parent herd in laying hens, causing reproductive dysfunction.

The effectiveness of ProStor for fish was shown in an experiment with young carp and sturgeon. The preparation in the amount of 2 kg per ton of compound feed was introduced into the compound feed KM-2M for carp and OT-7 for sturgeon. The yearlings were kept in aquariums in groups of 15 individuals for a month. Fish in the corresponding control groups received compound feed without the drug. The fish and biological indicators of juvenile carp and sturgeon at the end of the experimental feeding period in terms of absolute growth and average daily growth rate were higher in the experimental group compared to the control for carp, by 45% and 25%, respectively, and for sturgeons - by 120% and 45 % The effect of the drug was not detected on the safety of the carp population, both in the experiment and in the control, there was 100% survival, and in the group of experimental sturgeons the juvenile survival was 15% higher than the same indicator for the control fish.

Claims (1)

A method of obtaining a complex biologically active feed additive for farm animals, poultry and fish with probiotics and medicinal herbs, characterized in that they carry out separate deep cultivation of strains of Bacillus subtilis VKPM B-8130, Bacillus subtilis VKPM B-2984, Bacillus subtilis VKPM B-4099 and Bacillus licheniformis VKPM B-4162, obtaining their liquid cultures and subsequent solid-phase fermentation, for which the obtained liquid cultures of Bacillus subtilis VKPM B-8130, Bacillus subtilis VKPM B-2984 and Bacillus subtilis VKPM B-4099 are mixed in a ratio of 6: 6: 1, respectively, to irradiation of 65 l and applied to a pre-prepared carrier for carrying out solid-phase fermentation, sterile beet pulp in an amount of 200 kg, treated with a cellulolytic enzyme and enriched with fermentolizate of Saccharomyces cerevisiae fodder yeast, for which preparation: molasses - 255-265 g, potassium disubstituted - 98 - 98 102 g, magnesium sulfate - 25-26 g, fodder yeast Saccharomyces cerevisiae - 3 kg and water up to 30 l, the mixture is sterilized at a temperature of 120 ° C for 30 min and added to a carrier into which 4 l of a solution of celloviridine is added a or cellullox F with a cellulase content of at least 2000 units / g of carrier, adjust the pH to 6.0-6.5, mix thoroughly, incubate for 2 hours at a temperature of 45-50 ° C, and solid phase fermentation is carried out under conditions of limited oxygen availability at a temperature of 45-50 ° C, pH 7.5-8.0 and a batch humidity of 43-48% for 48-50 hours, 65 l of Bacillus licheniformis VKPM B-4162 liquid culture containing at least 5 are added to the resulting product, 6 · 10 ⁸ CFU / g, the mixture is thoroughly mixed and dried to a moisture content of 8-10%, after which dry powders of Echinacea purpurea herb and fruits are added. spotted ash at the rate of 20-50 g of echinacea powder and 20-50 g of milk thistle powder per 1 kg of the final product, the resulting mixture was stirred for 0.5 h and subjected to crushing to obtain a homogeneous mass.