RU2786910C1 - Method for the production of a functional feed product for farm animals - Google Patents

Abstract

FIELD: agricultural industry.

SUBSTANCE: invention relates to the agricultural industry, namely to fodder production. A method for the production of a feed product for farm animals, characterized in that it provides for the introduction of whey into the fermenter, the addition of nutrients to it and the introduction of prepared sawdust from birch or spruce in a ratio of sawdust:whey - 1:10, mixing, pasteurization, and after pasteurization, whey with added sawdust and nutrients is cooled to a temperature of 33±1°C, a Lactobacillus culture and a BK-Uglich-PRO preparation containing Propionibacterium freudenreichii are added, fermentation is carried out to reduce pH 4.0-4.6, then ultrasonic cavitation of the mixture is carried out when setting the output power of the generator corresponding to the intensity of ultrasound equal to 2.4 W/cm², then the mixture is cooled to 30±1°C and the top-fermenting fodder yeast Saccharomyces cerevisiae is introduced and cultivation is continued for 36-48 hours with aeration, cultivation is carried out to a concentration cells not less than 107 CFU in 1 ml, then sedimentation followed by permeate contamination, filtration and cooling of the biomass to 10°C.

EFFECT: invention makes it possible to obtain, in one technological cycle, an eco-friendly effective multifunctional feed product for farm animals using an ultrasonic extract of sugary substances from sawdust for growing protein fodder yeast on a nutrient medium with whey.

2 cl, 2 tbl, 2 ex

Description

The invention relates to biotechnology and can be used in the production of feed protein products using microbiological synthesis, which have a wide range of functional effects for farm animals associated with the composition of the product. The use of milk whey or dry whey as a component of the nutrient medium makes it possible to attribute the invention to the dairy industry. The use of ultrasonic sawdust hydrolyzate as the main source of carbohydrates makes it possible to attribute the invention to wood processing, specifically to the hydrolysis industry, namely, to an ecological method for obtaining hydrolyzates from wood for the subsequent biosynthesis of fodder protein.

It has been established by science and practice that the digestibility of vegetable proteins in farm animals does not exceed 80%, and the coefficient of its use in obtaining milk is not more than 30%. At the same time, in ruminants, the need for protein is satisfied by 20-30% due to the protein of rumen microorganisms, and 70-80% of the protein must be supplied with feed. The protein of the main plant foods is poor in essential amino acids: lysine, methionine, and tryptophan [1]. Feed accounts for more than half of all livestock production costs. The effectiveness of the functioning of the livestock industry, the level of competitiveness of finished products of domestic producers in the domestic and world markets and, ultimately, the quality of life of Russians will largely depend on the scientific justification and implementation of new developments in feed production [2].

In the diets of farm animals, fodder yeast is widely used as an additional source of complete protein. Feed yeast contains 3.0-3.5 g of lysine; methionine - up to 1.7%, tryptophan up to 1.3%. In most experiments, adding them (up to 25% of the protein requirement) to diets containing only vegetable proteins significantly increases the productivity of animals: the increase in meat, milk, wool increases, and in birds, egg production.

Feed yeast is a kind of natural premix consisting of a complex of essential amino acids, vitamins and the most valuable bioelements. Numerous practical studies prove that, in terms of zootechnical efficiency, fodder yeast is superior to meat and bone meal and slightly inferior to fish meal, even with its high quality [1]. At the same time, fodder yeast enzymes in terms of protease activity of pepsin are 2 times higher than meat and bone meal, 2.8 times higher than fish meal, 5-10 times higher than sunflower cake, barley and peas. The trypsin protease activity distinguishes yeast even more favorably from other feeds of animal and vegetable origin. At the same time, yeast activates trypsin in the gastrointestinal tract of animals. Thanks to these biological features, yeasts have a positive effect on the growth, development and productivity of animals. The shortage of feed protein in the world is estimated at about 30 million tons/year, and in Russia - at 2.3 million tons/year. At the same time, there is a close relationship between the food industry, biomedicine, and feed production, since the effective conversion of feed proteins in the body of productive farm animals ultimately provides the necessary level of human metabolism as the main consumer of the final product obtained from these animals [3].

Both plant mass [4] and oil and gas hydrocarbons [5] serve as sources of raw materials for obtaining biosynthetic feed protein. Plant sources of raw materials are currently of the greatest practical interest, because, firstly, unlike oil and gas hydrocarbons, they are constantly renewed through photosynthesis, and secondly, the mentality of the country's citizens at present in most cases does not allow the solution of all feed base issues with the help of petrochemical products.

Despite the fact that the processing of only 2% of the volume of oil produced annually makes it possible to produce up to 25 million tons of protein, which is enough to feed 2 billion people during the year, the development of the so-called environmentally friendly or organic animal husbandry still includes fodder products obtained using microbial fermentation of vegetable raw materials, not refined petroleum products. The volume of production of organic agricultural products in the world is growing every year. Organic agriculture has become a global trend, it is practiced in more than 160 countries. The total chemicalization of agricultural production and other anthropogenic factors have led to the fact that the system of ecological animal husbandry cannot be provided without the use of functional food additives in the diet of animals [4].

The first attempts to organize the industrial production of yeast for feed purposes were made at the beginning of the 19th century. A significant contribution to the development of the science of animal feeding was made by Russian scientists N. P. Chirvinskii, E. A. Bogdanov, M. F. Ivanov, E. F. Liskun, I. S. Popov, and others. I.S. Popov pointed out that dry fodder yeast is a good source of protein, vitamins, enzymes (proteases, nucleases, etc.).

In 1877, wood sawdust was hydrolyzed for the first time in Russia using a 1% solution of sulfuric acid, and the hydrolysis products (sugary substances) were used for yeast fermentation and the production of alcohol. Hydrolysis plants producing fodder yeast were launched in 1943-1944. at the Saratov and Khorsky plants. 1950-1980s In the mid 1980s. all hydrolysis plants became part of the Ministry of Medical and Microbiological Industry of the USSR. Many of them cooperated with sawmills, woodworking, pulp and paper and agricultural enterprises, which ensured an uninterrupted supply of raw materials. In those years, there were about 40 hydrolysis plants in the USSR located in various regions of the country, including Belarus, Kazakhstan, Moldova, Georgia, and Uzbekistan. On the territory of the Russian Federation, plants operated in the central and northwestern regions, in the Volga region, in the Urals, in Siberia and in the Far East. They produced more than 1 million tons of fodder yeast [6].

One of the methods for deep processing of wood raw materials is hydrolytic technologies, which result in the formation of wood hydrolysates containing soluble fractions of hexosans, pentosans, lignin, essential oils and mineral impurities. Sulfuric acid hydrolysis of both wood raw materials has gained an advantage, hydrolysis is known with solutions of hydrochloric, acetic and nitric acids, and alkaline hydrolysis using solutions of sodium hydrosulfite, sodium hydroxide and hydrolysis in a slightly acidic medium after alkaline treatment, as well as pre-treatment using an ionic liquid, peroxide hydrogen The process can be catalyzed by enzymes [7, 8, 9].

Lactic acid microorganisms and propionic acid bacteria are also important for the health of the normal microbiota of an animal. The modern development of fodder production is based on the introduction of resource-saving efficient biotechnologies of protein fodder yeast based on secondary production resources using selected consortiums of lactic acid and propionic acid bacteria [2, 10, 11].

Feed additives can contain not only 45-47% protein, but also a complex of biologically active substances, vitamins, amino acids, and sometimes live cultures of bacteria [2]. It is shown that propiovit, created on the basis of propionic acid bacteria, enhances the growth and productivity of poultry, piglets, calves by 2-12%, facilitates and accelerates the adaptation of animals to various stress factors. It is effective for the prevention of gastrointestinal diseases and for violations of mineral metabolism. Preparations based on acidophilus bacillus have a positive effect on egg production, egg quality and the microflora of the bird's digestive tract. A lyophilized culture of acidophilus bacillus is a preparation of biobacton. The use of biobacton increases the safety of hypotrophic piglets by 3-11%, growth rate by 5-7%, normalizes the main indicators of peripheral blood, helps to reduce the cost of purchasing feed. The use of a culture of lactic acid streptococcus increases the resistance of piglets to intestinal diseases, etc. There are studies on the effective use of other types of probiotics in animal husbandry. Many authors consider it promising to enrich the intestinal microflora of animals not with one culture, but with a number of specially selected strains, in other words, the use of complex probiotics [12].

The fodder base of agricultural enterprises should be created on the internal resources of farms, have the necessary reserves and at the same time include the cheapest sources of nutrients. Only under these conditions can the growing productivity of animals, the high quality of products and the consistent reduction in their cost be ensured. Wood raw material is characterized by rapid and continuous renewability, which is one of its most important advantages over most other raw material sources. Its average natural increase is estimated at 1.4 m ^{3 of} wood per 1 hectare of forest area [13].

Another highly valuable raw material for the biotechnology of obtaining a feed product for farm animals is whey, including dry whey [14]. It is known that the polluting capacity of 1 ton of whey exceeds that of domestic wastewater by more than 500 times, which is extremely dangerous for water bodies. The indicator of underutilization of whey varies in different regions, amounting to approximately 25%, while about 250 valuable compounds remain underused, including nitrogenous compounds, milk fat, minerals, lactose, vitamins, enzymes, and organic acids [17].

The presence in whey of easily digestible sources of carbon nutrition by many types of microorganisms, as well as various growth factors, makes it one of the most valuable nutrient media for obtaining microbial synthesis products. Approximately 20% of all known yeast species have the ability to assimilate lactose [15].

If it is impossible to quickly process whey, which is a perishable product, condensed or dry whey can be used. In condensed whey with a high concentration of solids, as a rule, there are no conditions for the development of most groups of microorganisms. In addition, condensed whey is more transportable and does not require large storage containers. Another product of this type is hydrolyzed condensed whey. Even less favorable are the conditions for the development of microflora in dry whey, which can be stored at positive temperatures for quite a long time, and the use of vacuum packaging increases this period by 4-5 times. Powdered milk whey is a biologically valuable product that, after reconstitution, can be successfully used as a nutrient medium in the process of microbial synthesis [16].

With additional enrichment of the environment with the missing bioelements for agricultural animals, it is possible to purposefully increase their content in a functional feed product.

In total, 60 genera and about 450 species of yeast are known. Some have found industrial use as a source of protein and other biologically active substances. The main representatives of the species Saccharomyces cerevisiae are used in the production of ethanol. The active form of Saccharomyces yeast is used in the baking industry, while the inactivated form is used as a source of protein, vitamins, etc. [17]. Good results in carbohydrate digestion and protein accumulation were shown by Saccharomyces cerevisiae of various races, strains of Rhodosporidium diobovatum, Torulopsis, Rhodosporidium diobovatum, Hansenula species, Debaryomyces hansenii, Guehomyces pullulans, Candida albicans, etc. [16, 17, 18].

Active catabolism of lactose is especially characteristic of yeasts from the genus Kluyveromyces, Debaryomyces hansenii, etc. The processing of whey by yeast cultures in order to obtain feed protein is a promising way to solve the problem of its utilization. It is also known that for the maximum accumulation of biomass on whey, it is necessary to enrich it with at least nitrogen, phosphorus, potassium and sulfur, for example, ammonium sulfate and potassium dihydrogen phosphate in the amount of 3.5 and 1 g/l, respectively [19]. Yeast accumulate mono and disaccharides (oligosaccharides) in its composition. Oligosaccharides are an integral part of the yeast wall. Therefore, the presence of such saccharides is beneficial for the digestion of animals and birds in terms of mycotoxin sorption. This means that yeast oligosaccharides should not be considered as their negative property. Organic acids are also stimulants of gastric digestion, contributing to the growth of protein digestibility in the diet of animals and poultry. These acids stimulate appetite and the growth of beneficial microflora in the intestines.

Often the value of fodder yeast is determined not only by the level of protein and amino acids in their composition, but also by the concentration of vitamins, a valuable source of which these products really are. Of particular value are the B vitamins accumulated by yeast cells in the course of their synthesis. Since vitamins are active sites of yeast cell synthesis enzymes, the accumulation of their concentration occurs in proportion to the growth of yeast cells. The cuticle, the shell of yeast, reliably protects vitamins from destruction during the drying of the feed product, and it also protects these substances from oxidation during yeast storage [20].

However, the development of yeast and the efficiency of protein accumulation are affected not only by the selection of the species, strain, race of yeast, but also by the composition of the substrate. When an additional carbohydrate source from wood hydrolysates containing hexose sugars is added to lactose or its hydrolyzate in whey, the intensity of protein biosynthesis increases [16].

Wood carbohydrates are mainly represented by polysaccharides, which make up about 70% of the wood substance (cellulose, hemicelluloses, starch, water-soluble polysaccharides), as well as low molecular weight sugars of juices and developing tissues. Cellulose and hemicelluloses dissolve in concentrated acids, and lignins in concentrated alkalis.

Coniferous species differ from hardwood in a significantly lower content of pentosans, their amount in hardwood wood is almost 2 times higher, but coniferous species contain a greater amount of lignin in their composition than in hardwood. Resin acids are also present mainly in softwood, and they are mostly extracted with hot water [27]. Until recently, there was no ideal process for obtaining microbial protein on cellulose substrates. A common disadvantage of processes of this type was the stressful ecology (substrate pretreatment, wastewater, dust and gas emissions, the difficulty of cleaning the obtained microbial protein from chemical reagents) [17].

In recent years, cavitation treatment using a low-frequency ultrasonic cell has been used to reduce the chemical intensity of the process of hydrolysis of wood waste.

Known ultrasonic technology for obtaining protein feed, which includes the feed ingredient is crushed and dispersed in water. Mineral raw materials, premixes, trace elements, vitamins and calcium-containing raw materials are added. Disintegrate by cyclic pumping of the resulting mixture in a closed circuit in cavitation mode at a temperature of 30-100°C. A dispersion medium is obtained by mechanical-hydropercussion-cavitation-dissipation processing. The mixture is pasteurized and adjusted for 20-120 cycles to a homogeneous state with a given particle size from 1 μm to 3 mm. The invention makes it possible to simplify the production of a biologically complete feed mixture and improve its absorption for cattle, pigs, poultry and fish [21]. The main disadvantage of this method is the mechanical composition of the nutrient mixture, which contains only ready-made ingredients and does not use the biotechnological potential of fodder yeast.

The following invention, related to the production of a feed probiotic preparation for farm animals [22], provides for the preparation of a phyton carrier by mixing fiber-containing plant materials, tap water, baker's yeast autolysate, dibasic potassium phosphate, magnesium sulfate and molasses in a given ratio of components. The resulting phyton carrier is sterilized and a liquid culture of Cellulomonas flavigena is added and kept at the specified parameters. Then sequentially make liquid cultures of Bacillus subtilis and Lactococcus lactis subsp. lactis in equal amounts. The resulting mixture is stirred, maintained at the specified process parameters and dried to a residual moisture content of 8-10%. At the same time, crushed straw, sunflower or soybean meal are used as fiber-containing plant materials, and liquid cultures of probiotic strains of Cellulomonas flavigena, Bacillus subtilis and Lactococcus lactis subsp. lactis is obtained by growing them on nutrient media designed for each strain of the mixture separately. EFFECT: invention makes it possible to increase the biological activity of the drug, to increase the digestibility of feed [22]. The disadvantages of this method are the complexity of obtaining the drug, the lack of significant nutritional value, allowing you to adjust the diet of animals in case of violation of its balance.

In other studies [23], when using enzyme-alkaline extraction with preliminary application of the method of explosive autohydrolysis of delignified cellulose samples, the increase in the reactivity of substrates after sonication was 52 and 65%, however, studies were carried out to obtain hydrolytic sugars in order to obtain biofuels.

In the method, which includes quotation treatment for obtaining a feed mixture, containing the preparation of a suspension in a rotary grinder-disperser from meal or cake and stillage at a temperature of 70-90 ° C, from which centrate enriched with waste grain production in a mixer is separated, characterized in that in crushed legumes are additionally added to the stillage and, together with grains, they are dispersed in a rotary grinder-disperser equipped with a cavitator for 7-10 minutes, and after centrifugation, the resulting suspension is kept for a day in the presence of cellulase to break down fiber to glucose or disaccharide in a container equipped with a bubbler , after which crushed bran and / or beer grains are introduced into the mixer and then excess moisture is removed from the prepared suspension, and drying is carried out until the moisture content in the finished product is not higher than 9 wt. % [24]. The method does not involve the use of whey and microbial biotechnology.

The level of technology and technology using whey is not characterized by the industry technological instruction RD-01.000.23-K.03-89 "Feeds and mixtures based on distillery stillage with improving additives", as in the above [24] method, according to the technical essence and the number of matching features, the Method for the production of a functional feed product for farm animals was chosen as the closest analogue [25].

The prototype [25] provides for a method for processing whey, including the additional introduction of nutrients into it, pasteurization of whey, fermentation of whey with bacterial cultures, one of which belongs to propionic bacteria, while during the cultivation process whey is aerated, whey is treated with cultures Lactobacillus casei and Propionibacterium freudenreichii, by a consortium of mesophilic fodder yeast, the ratio of yeast cultures to Lactobacillus casei is maintained from 1.0 to 5.0, cultivation is carried out to a cell concentration of at least 10 ⁷ CFU in 1 ml of Lactobacillus casei, Propionibacterium freudenreichii and yeast, then separated protein concentrate, which is a functional feed product for farm animals, by settling, followed by permeate contamination, filtration or separation on a cottage cheese separator.

Ready PCP is obtained in the form of a concentrate with a mass fraction of 47-50% or a dry powder with a mass fraction of solids of 9-10%, obtained by freeze-drying, which preserves the living cells of probiotic cultures of yeast races and strains of Propionibacterium and Lactobacillus.

Significant disadvantages of the prototype are: multi-stage, the need to use biopolymers of chitosan and pectin hydrocolloids, the complexity of the technological process, the parametric indicators of which (pH, concentration of cultures) under conditions of natural fluctuations in the chemical composition of the feedstock are difficult to implement, namely, that whey is sequentially treated with cultures of Lactobacillus casei and Propionibacterium freudenreichii, and the treatment with the second culture is carried out with a decrease in the pH value of whey from the initial value by at least 0.8 units, at the time of inoculation with the second culture, the ratio of cultures is maintained from 0.4 to 5.0, and the total concentration of cultures is chosen from 1 to 5 g/l of medium; a significant disadvantage is also that the content of carbohydrates in whey, insufficient for the development of yeast (approximately 5% lactose, including partially hydrolyzed into glucose and galactose), inhibits the development of the accumulation of protein mass.

The development of yeast and the course of fermentation are primarily influenced by the following factors: sugar concentration, temperature, acidity of the environment and the presence of trace elements, oxygen access. Cultural yeasts are acidophiles, that is, they develop in an acidic environment, the optimal pH value for yeast is 4.5–5.0. Under aerobic conditions, they actively grow and multiply, and in anaerobic conditions they carry out alcoholic fermentation, for the accumulation of protein it is necessary to avoid anaerobic conditions. To avoid growth retardation to increase the yield of the product, a concentration of sugars of 10-15% is provided in the nutrient medium [26].

The aim of the invention is to obtain in one technological cycle an environmentally friendly effective multifunctional feed product for farm animals using an ultrasonic extract of sugary substances from sawdust for growing protein fodder yeast on a nutrient medium with whey.

To achieve the technical result provided by the invention, milk whey, or reconstituted condensed whey or reconstituted whey powder, is additionally supplemented with nutrients, including carbohydrates, which are sugary substances obtained by low-frequency ultrasonic cavitation from sawdust, fermentation of whey is carried out with cultures of probiotic bacteria, while in the process of cultivation, whey is aerated, then the protein concentrate, which is a functional feed product for farm animals, is separated by settling, followed by permeate contamination, filtration, separation, or the protein concentrate is not separated from a part of unfermented carbohydrates, which contain microcellulose and a functional feed product for farm animals.

At the same time, the technology for obtaining sugary substances from sawdust is combined with the technology for fermenting whey and consists in the claimed method for the production of a functional feed product for farm animals as follows:

Bioelements nitrogen, potassium, phosphorus and sulfur are added to milk whey with a content of (5.5 + 0.5)% dry matter (or restored from dry or concentrated whey to this dry matter content), if necessary, enrichment of the finished product with additional bioelements selected from copper, iron, manganese, magnesium, zinc, cobalt, iodine, selenium and prepared wood-shaving raw materials, crushed to a size of not more than 3 mm, in a ratio of whey with components: sawdust - 10:1 by weight. Next, whey is pasteurized with components, including sawdust at (75 ± 1) ° C for 2 minutes and cooled to (33 ± 1) ° C, the introduction of cultures of probiotic strains of lactic acid cultures, at least of the genus Lactobacillus and Propionibacterium and, cultivation at (33 ± 1) ° C until a pH of 4.0-4.6 is reached, during this fermentation sawdust swelling occurs at the same time, after which a low-frequency receipt processing of the mixture is carried out (hereinafter, the mixture after cavitation treatment is the substrate), subsequent introduction into fodder yeast culture substrate and fermentation at (33±1)°C, cultivation at the specified temperature for 24-48 hours with aeration, until culture biomass is at least 10 ⁷ CFU/ml; cooling of the protein-yeast biomass to 10°C, settling followed by contamination of the permeate, filtration or separation on a separator-curd separator to a mass fraction of solids of 47-50%, or without separating the permeate, followed by drying of the fermented substrate on a lyophilic, spray or roller dryer to obtain a dry product.

Ready PCP is obtained in the form of a concentrate with a mass fraction of 47-50% or a dry powder with a mass fraction of solids of 8-10%, obtained by freeze-drying, which preserves the living cells of probiotic cultures of yeast races and strains of Propionibacterium and Lactobacillus, or by drying, selected from spray and roller, in which living cells of probiotics are not preserved, but when using probiotic cultures with metabiotic properties, metabolites are preserved that have a beneficial effect on animal health, in particular, have an antibacterial property, which is a safe alternative to feed antibiotics in ecological animal husbandry [27] .

In continuous industrial cultivation or a batch process with feeding of the substrate, providing a stationary phase of growth of probiotic cultures.

Comparison of the proposed invention with other technical solutions known from the prior art made it possible to establish the following differences related to the technical solution:

- the proposed method allows to reduce wood waste and underused whey, from which a multifunctional feed product for farm animals is obtained in one technological cycle: an ultrasonic extract of sugary substances for growing protein fodder yeast, a protein feed product, bioelements and a complex of other biologically active substances passing from raw materials and formed in the process of fermentation during the cultivation of probiotic cells in vitro, live probiotic cultures and / or metabolic products of these cultures;

- the products of wood hydrolysis are extremely diverse, among them more than 200 individual compounds have been isolated that are formed as a result of the destruction of individual components of wood; during the hydrolysis of some types of wood, substances are formed that can inhibit the development of yeast and other probiotic crops, so we additionally conducted studies on the content of sugar substances in ultrasonic extracts of various types of wood and assessment of the toxicity of impurities in them by a biotechnological method; birch and spruce wood showed the best indicators of non-toxicity for ciliates and microorganisms;

- in the composition of spruce, unlike birch, there are up to 30.1 extractive substances [27], which can give a taste of bitterness, but most of them are extractable with hot water, so the preparation of spruce wood requires not only its grinding, but also the extraction of bitter compounds , which may be toxic to some probiotic cultures;

- whey has a high nutritional value and contains, in addition to whey proteins, milk fat and a number of bioelements, on average, mg: calcium - 103, magnesium - 10, phosphorus - 78, potassium - 143, sodium - 48, zinc - 0.4, selenium - 1.8, wood also, in addition to the high carbohydrate component, contains, with some fluctuation, up to 1% of bioelements, which, when used, can ensure the usefulness of the nutrient medium for the crops used in its fermentation, with the exception of nitrogen, potassium, phosphorus, sometimes sulfur , the doses of which are reduced compared to a medium that does not contain milk whey, or cavitation hydrolyzate of sawdust;

- the use as a raw material at the same time: whey, and sawdust, containing in total on average the necessary minimum of nutrients for the development of used probiotic cultures, allows to reduce the amount of added bioelements qualitatively and quantitatively, to use in the minimum variant for enriching the substrate only sources of nitrogen, potassium and phosphorus; in the maximum variant, additionally those bioelements, the lack of which is established in animals for which the functional feed product is intended;

- from the patent and scientific and technical literature, there is no known method for the production of an extract of sugary fractions of wood waste without chemical reagents, obtained from sawdust of spruce and birch using a low-frequency ultrasonic reactor, which provides deep penetration of the solvent into the structure of wood particles and, due to cavitation effects, causes an increase in speed hydrolysis of wood components, the resulting sugary fractions from wood waste are used to prepare a substrate based on whey, or a reduced version of whey for subsequent fermentation of the substrate with fodder yeast cultures;

- in the EU countries, livestock farms have been banned from using antibiotics for prophylactic purposes, which follows from the updated EU Regulation "On Veterinary Medicinal Products", which entered into force on January 28, 2022, however, probiotic cultures with antibacterial and metabolic properties, for a long time used in food biotechnologies are safe and can be used in the production of organic livestock products [28]. Probiotics and prebiotics are used in modern animal and poultry production to stabilize the intestinal microflora in monogastric animals and the rumen environment in ruminants. Especially after the ban on the use of antibiotics as growth promoters in the European Union, the use of pro- and prebiotics in animal nutrition is becoming increasingly important. Preparations, which include microcrystalline cellulose, hemicellulose, lignin, collectively related to insoluble fiber, belong to the group of herbal medicines with prebiotic properties.

The method for processing whey, which implements the proposed technical solution, is an additional introduction of nutrients into the whey or reconstituted whey and the introduction of prepared sawdust in the ratio of sawdust:whey - 1:10, mixing, pasteurization, and after pasteurization of the whey with the added sawdust and its nutrients cooled to a temperature of (33 + 1) °C, cultures of Lactobacillus and Propionibacterium are added, fermentation is carried out until the pH decreases to 4.0-4.6, then ultrasonic cavitation is carried out when the output power of the generator is set, corresponding to the intensity of ultrasound, equal to 2.4 W/cm ² , then fodder yeast cultures are added and cultivation is continued with aeration, cultivation is carried out to a cell concentration of at least 10 ⁷ CFU per 1 ml of Lactobacillus, Propionibacterium and yeast, the finished protein product is obtained in the form of a concentrate or dry powder. When using a drying method that does not ensure the survival of an effective amount of live probiotic cultures, in particular spray or roller drying, at least one strain with known metabiotic characteristics is used to obtain a product with metabolic properties [29, 30, 31].

To achieve the specified technical result is carried out sequentially:

- acceptance of whey with a mass fraction of solids of 5.0-5.5% (curd, cheese, casein) in accordance with GOST R 53492-2009, or recovery of condensed / dry whey (GOST 33958-2016 Dry milk whey. Specifications ) to a similar mass of dry matter, which is sent to fermenters with the functions of sterilization, cooling and aeration;

- adding to the fermenter bioelements, prepared crushed sawdust of spruce or birch, mixing and pasteurization of whey at (75±1)°C for 2 minutes;

- cooling to (33 + 1)°C, adding probiotic cultures, including at least Lactobacillus, Propionibacterium and cultivating them to pH 4.0-4.6;

- then the mixture of sawdust with serum and probiotic cultures is subjected to ultrasonic cavitation when the output power of the generator is set, corresponding to the intensity of ultrasound, equal to 2.4 W/cm ² , cooling to (33±1)°C;

- adding a fodder yeast culture and fermentation is carried out at (33±1)°C, while aeration is carried out during the cultivation of cultures;

- cultivation at the specified temperature for 36-48 hours with aeration, until the biomass of cultures is not less than 10 ⁷ CFU/ml;

- biomass cooling down to 10°C;

- it is possible both to settling with contamination of the permeate of the upper part, filtration or separation on a separator-curd separator to a mass fraction of solids of 47-50%, and the use of the resulting biomass without separation for subsequent drying.

Ready PCP is obtained in the form of a concentrate with a mass fraction of 47-50% or a dry powder with a mass fraction of solids of 9-10%, obtained by freeze-drying, which preserves the living cells of probiotic cultures, roller or spray drying, in this case cultures with a metabolic properties.

Probiotic cultures, which are obtained by growing them on nutrient media intended for each strain of the mixture separately, or activated, bacterial concentrates containing these cultures, in accordance with the manufacturer's recommendations.

When choosing probiotic cultures of yeast races and strains of Propionibacterium and Lactobacillus, they are guided by research data:

- their probiotic potential at different levels: specific antagonism in relation to pathogenic and opportunistic microflora;

nonspecific antagonism in relation to pathogenic and conditionally pathogenic microflora; cellular and humoral immunity, as well as in vitro biocompatibility of probiotic microorganisms with indigenous strains of the normobiota of specific animals for which the product is intended, etc. [26];

- improvement of controlled indicators of normalization of metabolism, acid-base balance, and other indicators of animal health and development [27];

- ensuring a low concentration of purine and pyrimidine residues of nucleic acids in the product;

- the amount of the product to be fed is set in such a way that its introduction into the diet serves as a preventive measure to ensure the health of the animals;

- the metabolic properties of the strains used, which allows in functional feed products to simultaneously increase the shelf life of the product, and provide probiotic properties due to metabolites, because the bactericidal activity of such strains is up to 40% higher in lysates than in live cultures [28,29,30];

- the content of polysaccharides (cellulose, hemicellulose, pectin) in the finished product, necessary and safe for a given animal species, as well as a polyphenolic substance - lignin, determined by a modern detergent analytical system [32, 33, 34].

The production of a feed product according to this method is carried out in the conditions of both small and large workshops for the processing of whey of biotechnological industries, and it is also possible directly in livestock farms.

Example 1. A concentrated multifunctional feed product produced directly in small and medium-sized livestock farms with an established deficiency of the manganese bioelement in animals.

Batch fermenters fill 1/3-1/2 part with whey, add (by weight) 0.5% urea, 0.1% potassium hydrogen orthophosphate, 0.04% manganese dioxide, 10% sawdust crushed to a state of flour, for example , after the belt sander; is mixing, pasteurization at (75±1)°C for 2 min; cooling down to (33+1)°С; adding probiotic cultures, fermenting to pH 4.0-4.6, sonicating a mixture of sawdust with whey and probiotic cultures with ultrasound when setting the output power of the generator, corresponding to the intensity of ultrasound, equal to 2.4 W/cm ² ; cooling to (30±1)°C, adding a fodder yeast culture and fermenting at (30±1)°C, while aeration is carried out during the cultivation of cultures;

cultivation at the specified temperature for 36-48 hours with aeration until the culture biomass reaches at least 10 ⁷ CFU/ml, sedimentation followed by permeate contamination, filtration and cooling of the biomass to 10°C.

A preservative (in particular, sodium citrate) can be added to the resulting protein concentrate, followed by packaging in sealed containers, or the concentrate is dried in any way that ensures the claimed functionality of the product.

In the given example, Lactobacillus acidophilus 22 ₂ w, viscous as part of the activated bacterial preparation BK-Uglich-AV and bacterial concentrate of propionic acid bacteria, BK-Uglich-PRO, and the preparation of active top-fermenting dry yeast Saccharomyces cerevisiae were used as probiotic cultures. Before use, the concentrates were dissolved in sterile water heated to 30–40°C at the rate of 1 sachet (1 EA) per 50–100 ml of liquid and kept for 25–30 min to swell.

The composition of the product protein concentrate according to example 1:

- JBU: protein according to Bernstein (in terms of absolutely dry matter), not less than 32%; crude fiber - 1.3-2.7%; crude fat - 2.7-3.3%;

- dietary fiber - 2.9 g / kg.

The multifunctionality of the feed product obtained according to method 1 includes at least: high protein content, prebiotic in the form of insoluble fiber (cellulose, hemicellulose, lignin) and oligosaccharides, manganese, live probiotic cultures.

The Lactobacillus acidophilus 22 ₂ w strain is known for its high level of antibacterial activity against Staphylococcus aureus, Micrococcus flavus, Proteus vulgari, Aspergillus niger, etc., inhibiting the growth of pathogenic bacteria and micromycetes [30].

Manganese is essential for the normal reproduction of both males and females and is a cofactor for a number of enzymes that regulate carbohydrate metabolism.

Example 2. Dry multifunctional feed product to compensate for the deficiency of protein, zinc, cobalt, iodine and selenium, in the absence of iron hypoelementosis, produced in a specialized biotechnological production workshop.

Feed yeast of a stable composition and quality is obtained when the technological process is not interrupted, and the production volume exceeds 40-50 tons of the finished product per shift [20]. At large plants with established proper control over the technological process and the quality of the finished product, with minimal fluctuations in the effectiveness of probiotics, yeast from batch to batch, it is possible to comply with the standard for ensuring the content of TBU, amino acids, a number of vitamins (thiamine B ₁ , riboflavin) in the finished multifunctional feed product B ₂ , pantothenic acid B ₃ , choline B ₄ , pyridoxine B ₆ , nicotinic acid, biotin H, folic acid B _c , cyanocobalamin B ₁₂ , tocopherol E), bioelements (calcium, phosphorus, sodium, potassium, magnesium, chlorine, iron , zinc, copper, manganese, cobalt, iodine, selenium), etc.

The production process begins with the preparation of raw materials.

Crushed spruce sawdust is poured with hot water at a temperature of (95 ± 1) ° C in the ratio of sawdust: water 1:10, kept for 2-3 hours to swell at a temperature not lower than 40 ° C, then the mixture of sawdust with water is subjected to ultrasonic cavitation when setting the output the power of the generator, corresponding to the intensity of ultrasound, equal to 2.4 W / cm ² withstand the mixture of sawdust subjected to cavitation with water for 2-3 hours to complete the extraction, filter and consider the sawdust washed from extractive substances prepared for the next stage of production. The finer the sawdust is crushed, the less exposure is needed for their swelling and extraction of resinous substances from them.

Continuous fermenters fill 1/2-2/3 parts with condensed cheese whey or dry milk whey restored to 5.0-5.5% of solids, add mineral components that can be adjusted in qualitative and quantitative terms depending on the established need animals for which the product is intended, in particular (g/l), table 1:

Table 1.

mineral component Bioelement content, % Addendum
into serum Ammonium sulfate 21% nitrogen; 24% sulfur 3.2 g/l Potassium hydrogen phosphate 17% phosphorus; 44% potassium 2.0 g/l zinc sulfate 40% zinc; 19% sulfur 786 mg/l Copper sulfate (II) pentahydrate 25% copper; 12% sulfur 177.60 mg/l potassium iodide 23% iodine, 21% potassium 16.52 mg/l Cobalt(III) sulfate
eighteen-water 16% cobalt; 12% sulfur 24.37 mg/l sodium selenite 45% selenium 3.33 mg/l

Additional introduction of nutrients into whey includes the introduction of prepared sawdust in the ratio of sawdust:whey - 1:10, mixing.

Next, the whey is pasteurized at (75+1)°C for 2 minutes; cooled to (33 + 1) ° C, cultures of probiotic lactic and propionic acid bacteria are introduced:

1) BIFILACT-PLUS containing Lactobacillus plantarum, Lactococcus lactis subsp. Diacetilactis, Streptococcus thermophilus (viscous), Bifidobacterium bifidum and/or B. longum and/or B. adolescentis;

2) BK-Uglich-PRO containing Propionibacterium freudenreichii

and fermentation is carried out until the pH decreases to 4.0-4.6, then ultrasonic cavitation when the output power of the generator is set, corresponding to the intensity of ultrasound, equal to 2.4 W / cm ² , cooling to (30 + 1) ° C, then fodder yeast cultures are introduced into the substrate .

Further cultivation is carried out for 36-48 hours with intensive aeration, maintaining the level of dissolved oxygen at least 10% of saturation. With continuous industrial cultivation, the stationary phase of growth of probiotic cultures is provided by adding a substrate. Then the biomass is cooled, freeze-dried.

The multifunctionality of the feed product obtained according to method 2 includes at least: a high protein content, live probiotic cultures and a guaranteed supply of bioelements in a functional dose of at least 15% of the established average daily norm (the calculation was made when feeding 100 g of a dry product with a mass fraction dry matter 90% for the need for nutrients for dairy cows with a milk yield of up to 8000 kg of milk (fat content in milk 3.8-4.0%, protein 3.2%, live weight 600 kg, 5 months of lactation) [ 35], table 2.

Table 2. Bioelement composition of the enriched functional product for cattle

micronutrient Norm Content in 100 g of lyophilized
lump product Average security
from the daily norm Potassium 24% Phosphorus 16% Sulfur 24% Zinc 1429 mg 314.4 mg 22% Copper 222 mg 44.4 mg twenty% Iodine 20 mg 3.8 mg nineteen% Cobalt 17.8 mg 3.9 mg 21% Selenium 6.3 mg 1.5 mg 24%

Sources of information:

1. Kalyunyants K.A., Ezdakov N.V., Pivnyak I.G. Application of products of microbiological synthesis in animal husbandry / K.A. Kalunyants, M.: Koloss. 1980. - 228 p.

2. Volkova G.S., Rimareva L.V., Arifullina L.R. Bioconversion of secondary raw materials of alcohol production into protein feed additives / Perspective enzyme preparations and biotechnological processes in food and feed technologies. M.: State Scientific Institution of the Research Institute of Food Biotechnology of the Russian Academy of Agricultural Sciences. 2016. S. 275-282.

3. Grishin D.V., Podobed O.V., Gladilina Yu.A. Bioactive proteins and peptides: current state and new trends in practical application in the food industry and fodder production // Problems of nutrition. 2017. V. 86. No. 3

4. Fish meal must comply with the Russian agricultural portal https://agroportal-ziz.ru/articles/rybnaya-muka-dolzhna-sootvetstvovat-gostu

5. Donnik I.M., Voronin B.A. Production of organic livestock products in the Russian Federation // AVU. 2016. No. 5 (147). URL: https://cyberleninka.ru/article/n/proizvodstvo-produktsii-organicheskogo-zhivotnovodstva-v-rossiyskoy-federatsii-normativno-pravovoe-

6. Nesmeyanov A., Belikov V. Food of the future. https://sheba.spb.ru. Moscow: Publisher: Pedagogy. 1985.

7. Sukhodolov A.P., Khamataev V.A. Development of the domestic hydrolysis industry. Izvestiya BGU. 2009. №3. URL: https://cyberleninka.ru/article/n/razvitie-otechestvennoy-gidroliznoy-promyshlennosti.

8. Golyshkin, A. V. Comparison of various types of chemical pretreatment of wheat straw and beech sawdust // Young scientist. - 2016. - No. 27 (131). — S. 14-18. — URL: https://moluch.ru/archive/131/36607/

9. Privalova Elena Andreevna, Fomina Elena Sergeevna, Evstafiev Sergey Nikolaevich Influence of the component composition of straw on the rate of enzymatic hydrolysis of cellulose // Bulletin of ISTU. 2010. No. 7 (47). URL: https://cyberleninka.ru/article/n/vliyanie-komponentnogo-sostava-solomy-na-skorost-fermentativnogo-gidroliza-tsellyulozy (Date of access: 01/27/2022).

10. Zeltina M, Leite M, Apine A., Shvinka Yu. et al. Obtaining a feed protein preparation from straw and other low-value agricultural and forestry waste // Biotechnology & BioE. No. 5, 1991. S. 4-5.

11. Efimova, L.V. Effective microorganisms in feeding cattle and pigs /L.V. Efimova, T.A. Udalova; Krasnoyarsk NIIZh of the Russian Agricultural Academy. - Krasnoyarsk. 2011.

12. Feed biotechnology: a textbook for students in areas of training 35.03.03 Agrochemistry and soil science, 35.03.07 Technology of production and processing of agricultural products, 35.03.04 Agronomy. FGBOU VPO Primorskaya State Agricultural Academy / FGBOU VPO PGSHA; comp. E.P. Ivanova, O.M. Puffer. - Ussuriysk, 2015. - 92p.

13. Tarasov S. M., Kononov G.N. Complex chemical processing of wood. Technology of wood-chemical and hydrolysis industries. Educational and methodical manual for students of the direction of preparation 18.03.01, 18.04.01. - M.: FGBOU VO MGUL, 2016. - 122 p.

14. Sokolenko G.G., Ponomareva I.N., Elizarova T.I., Esaulova L.A. Biotechnology of yeast whey product // Vestnik OrelGAU. 2016. No. 5. URL: https://cyberleninka.ru/article/n/biotehnologiya-drozhzhesyvorotochnogo-produkta

15. Zipaev D. V., Zimichev A. V. Milk whey is a valuable raw material for secondary processing. Izvestiya vuzov. Food technology. 2007. No. 2. URL: https://cyberleninka.ru/article/n/molochnaya-syvorotka-tsennoe-syrie-dlya-vtorichnoy-pererabotki

16. Le Anh Tuan “Application of psychrotolerant yeast for bioconversion of secondary resources of vegetable raw materials processing. Dissertation for the degree of candidate of technical sciences. - Kazan, KNRTU, 2016. - 167 p.

17. Ruchay, N. S. Ecological biotechnology: a textbook for students of the specialty "Bioecology" of universities / N. S. Ruchay, R. M. Markevich. - Minsk: BSTU, 2006. 311 p.

18. Turshatov M. V., Kononenko V. V., Ledenev V. P., Krivchenko V. A., et al. Technological bases for obtaining protein feed products during the processing of starch-containing raw materials into biotechnological and chemical products // Storage and processing of agricultural raw materials. 2018. №2.

19. Ilyushchenko A.F., Yakimovich N.I., Yakimovich I.V. Obtaining fodder protein by processing whey with yeast cultures. Modern problems of development of new equipment, technologies, organization of technical service. pp. 296-272. https://rep.bsatu.by/handle/doc/4748

20. Podobed L.I. Feed yeast - comparative characteristics of nutritional and biological value. 2007. http://podobed.org/kormovye_drozhzhi_-_sravnitelnye_harakteristiki_pitatelnoy_i_biologicheskoy_tsennosti.html

21. Senichev, V. P. (2015) Efficiency of ultrasonic treatment in the process of extracting reducing sugars in crushed wood / V. P. Senichev, L. M. Voropai, Yu. mezinárodní vědecko-praktická conference, Prague-2015. – Prague, 2015.

22. Pat. 2447674 RF (2010), MPK51 S23K 1/00. A method for obtaining a biologically complete feed mixture / Sidorov A.V., Kovalev A.V., Moshkutelo I.I.; applicant and patent holder Limited Liability Company "Kovikorm" No. 2010135931/10; dec. 08/31/2010; publ. 04/20/2012. – B.i. – 2012 No. 11.

23. Effect of ultrasonic cavitation on the reactivity of lignocellulosic substrates during the bioconversion of plant biomass

Dontsov A. G., Karmanov A. P., Kocheva L. S., et al. Butlerov's communications. 2014. V.39. No. 9. pp. 52-57.

24. Method for preparing protein feed mixtures No. of title of protection RID 0002615814 04/11/2017. Kovalev A.V., Sidorov A.V. LLC "Kavikorm Engineering"

25. Method for the production of a functional feed product for farm animals Patent for invention RU 2652155 C1, 04/25/2018.

26. Suchkova E.P., Uvarov R.A., Fundamentals of biotechnology - St. Petersburg: ITMO University, 2021. - 83 p.

27. Tarasov S. M., Kononov G.N. Complex chemical processing of wood. Technology of wood-chemical and hydrolysis industries. - M.: FGBOU VO MGUL, 2016. 122 p.

28. GOST 33980-2016 Interstate standard. Products of organic production.

29. Polyanskaya I, Stoyanova L, Popova V. Сoncept of metabiotics in fermented dairy products // JHED December Volume, December 2021.

30. Polyanskaya I, Shigina E, Popova V. Metabiotics fermented milk product // JHED December Volume, December 2021.

31. Stoyanova L.G., Dbar S.D., Polyanskaya I.S. Metabiotic properties of lactobacillus acidophilus strains included in complex starter cultures for the production of probiotic dairy products // Biotechnology. 2022.

32. MU 13-7-2/2156. Guidelines for the accelerated determination of the toxicity of animal products and feed.

33. GOST 31674-2012. Feed, compound feed, compound feed raw materials. Methods for determining general toxicity

34. Fundamentals of nutrition and feeding of farm animals: educational and practical guide / V.G. Ryadchikov - Krasnodar: KubGAU, 2012. - 328 p.

35. Golovin A.V. Influence of carbohydrate-protein ratio in diets on the productivity and metabolism of highly productive cows. Vologda: SZNIIMLPH, 2020.

Claims (2)

1. A method for the production of a feed product for farm animals, characterized in that it provides for the introduction of whey into the fermenter, the addition of nutrients to it and the introduction of prepared sawdust from birch or spruce in the ratio of sawdust:whey - 1:10, mixing, pasteurization, and after pasteurization, the whey with added sawdust and nutrients is cooled to a temperature of 33 ± 1 ° C, a Lactobacillus culture and a BK-Uglich-PRO preparation containing Propionibacterium freudenreichii are introduced, fermentation is carried out until the pH decreases to 4.0-4.6, then ultrasonic cavitation of the mixture when setting the output power of the generator, corresponding to the intensity of ultrasound, equal to 2.4 W/cm ² , then the mixture is cooled to 30±1°C and the top-fermenting feed yeast culture Saccharomyces cerevisiae is introduced and cultivation is continued for 36-48 hours with aeration, cultivation carried out to a cell concentration of at least 10 ⁷ CFU in 1 ml, then settling followed by contamination of the permeate, filtration and cooling of the biomass to 10°C. 2. A method for producing a feed product for farm animals according to claim 1, characterized in that milk whey, or reconstituted condensed milk whey, or reconstituted dry milk whey is used as whey.