RU2721795C1 - Composition and method of producing biocomposite fodder additive for farm animals and poultry - Google Patents

Abstract

FIELD: agriculture.

SUBSTANCE: group of inventions relates to agriculture, in particular to composition of biocomposite fodder additive for farm animals and poultry and a method for production thereof. Composition contains montmorillonite-containing clay enriched with 10 % hydrochloric acid and biomass of fodder yeast selected from Pichia pastoris, Faex medicinalis, Saccaromicete spp or combinations thereof, with following ratio of initial components in wt%: montmorillonite-containing clay enriched and activated with 10 % hydrochloric acid 10–20, biomass of fodder yeast 90–80. Method of obtaining is characterized by pre-sedimentation enrichment and activation of montmorillonite-containing clay with hydrochloric acid of 10 %, then washed with distilled water purified to neutral medium, dried at a temperature of not more than 105 °C and milled until homogeneous consistence. Biomass of fodder yeast selected from Pichia pastoris, Faex medicinalis, Saccaromicete spp or a combination thereof, is filled with distilled water, mixed to a homogeneous mass and treated in an ultrasonic generator at 1,500 Hz 4 times for 10 minutes, cooling system for 5 minutes after each treatment, then, the biomass treated with yeast is centrifuged, the supernatant is drained, the biomass residue is heated to 40 °C and added to the enriched montmorillonite-containing clay at a constant ratio under constant mechanical stirring. Mixture is thoroughly stirred for at least 40 minutes, then subjected to soft drying at t=65 °C to moisture content of not more than 30 % and milled to finely dispersed homogeneous medium.

EFFECT: using the group of inventions enables preventing gastrointestinal diseases and intoxication of various aetiologies.

3 cl, 1 tbl, 6 ex, 1 dwg

Description

The composition and method of producing a biocomposite feed additive for farm animals and poultry relates to agriculture, namely the feed industry, and can be used to prevent diseases of the gastrointestinal tract and intoxications of various etiologies in farm animals and poultry by binding mycotoxins.

One of the most important factors in increasing the productivity of farm animals and poultry is to provide the economy with high-quality feed. Low-value coarse feed, waste from the food and agricultural industries, are used as a feed base, which saves high-quality grain. But at the same time, it is roughage that most often undergoes rotting, mold microflora, exhaust gases, residual chemicalization from fertilizers, etc. As a result of such phenomena, conditionally pathogenic and pathogenic microorganisms, toxic waste products of fungi — mycotoxins, heavy metal ions, radionuclides, half-life products of organic phenols, which negatively affect the health of the animal or bird, get into the feed. Thus, the presence of a sorbent as a biological additive for feeding animals with coarse feeds is necessary.

Currently, various natural mineral sorbents are used, which are subsequently crushed in crushers, ball, planetary and vibratory mills to achieve small particle sizes in order to increase the sorption surface.

Known antibacterial properties of montmorillonite containing clay deposits of the Belgorod region, which in concentrations of 12.5; 25-50 and 50-100 mg / ml inhibit the growth of Escherichia coli and Salmonella enteritidis (Bukhanov V.D., Vezentsev A.I., et al. Antibacterial properties of montmorillonite-containing sorbents // Scientific statements of BelSU. Series of Natural Sciences. - 2011. - No. 21 (116) - Issue 17. - S. 57-63). It has also been proven that the combined use of modified montmorillonite-containing clay with entrofloxacin and thymol provides local detoxification, has an antiadhesive effect on the pathogenic intestinal microflora, and thereby reduces the treatment time for sick animals (Zuev N.P., Bukhanov V.D. et al. Efficiency of composite preparations based on nanostructured montmorillonite-containing clays in bird escherichiosis // Innovations in the agricultural sector: problems and prospects. - 2018. - No. 1. (17). - P. 169 - 175.).

Known feed additive for lactating cows, which consists of bentonite clay, salt, precipitate, magnesium oxide. The additive is prepared by mechanical mixing and fed depending on the daily milk yield. The feed additive can be fed not only with concentrated feed, but also with silage, haylage and green mass (RF patent 2081611 from 1997.06.20)

Known feed additive to increase the resistance and productivity of farm animals and birds (Patent No. 2294648 from 2007.03.10), which includes oregano essential oil (oregano) and bentonite in the following ratio, wt.%: Oregano essential oil (oregano) 3.0 -7.0; bentonite rest. The feed additive is highly effective and improves the resistance and productivity of farm animals and poultry.

Known feed additive "Celeben" (Patent No. 2432774 dated 2011.11.10), including diacetophenonyl selenide and bentonite in a ratio of 8 g to 13 g bentonite per 1 mg - 3 mg diacetophenonyl selenide, using bentonite from the Tarn-Var deposit of the Republic of Tatarstan. Using the invention allows to activate protein, carbohydrate, mineral and vitamin metabolism, increase the amount of total protein and glucose in the blood, increase the body's natural resistance by enhancing the phagocytic activity of leukocytes. Feeding supplements enhances reproductive function in animals, provides an increase in live weight gain in animals, animals and birds, as well as the safety of the livestock.

Known mineral-fat feed additive for cattle (Patent No. 2500174 dated 2013.12.10), characterized in that it includes the following components,%: bentonite of the Donguz deposit - 18.5, fusal sunflower - 13.3, sodium selenite - 0.002 wheat bran - 8.498, electroactivated water - catholyte with a pH of 9.0-9.5 and a redox potential of 400-500 mV - 59.7. In the invention, it is presented that in the carcasses of experimental gobies the groups of meat of the highest grade were more by 0.5 and 1.8 kg (1.9; 9.1%), the first grade by 1.8 and 6.5 kg (1.9 ; 7.1%). However, the introduction of sunflower into the fusa will block the sorption and adhesive properties of bentonite in the digestive tract of animals.

Known feed additive for farm animals and poultry (Patent No. 2579219 dated 2016.04.10), containing bee subpestilence, biomass and culture fluid obtained from cultivation of Medusomyces Gisevii Lindau Kombucha, bifidobacterin, pine and aspen bark extract, flask, bentonite and grass meal . The use of the invention will stimulate metabolic processes, as well as remove heavy metals and mycotoxins from the body of animals. The disadvantage is that it has a complex composition, including significant amounts of valuable medicinal substances, such as pine bark and aspen extract, flask, as well as bifidobacterin, which increases the cost of the composition.

In the practice of feeding in the Russian Federation, the domestic complex preparation Fungistat is widely used. From the description of the RF patent No. 2420565 (published on June 10, 2011), the method of producing the biological product “Fungistat” for eliminating mycotoxins from food and feed raw materials involves mixing in a ratio of 5: 1 a sorbent selected from aluminosilicates and a sorbent selected from layered sorbents, the compound obtained mixtures with riboxin, lecithin, L-carnitine, an organic acid selected from the group consisting of succinic acid and propionic acid, or a salt thereof, a mixture of oligofructose and inulin (30:70) and an enzymatic proteolytic preparation based on Bacillus subtilis bacterial culture fluid.

Known feed additive "Carbitox" (Liman ES, Reznichenko L.V. The effectiveness of the sorption capacity of Carbitox in relation to various mycotoxins // Scientific notes of the Kazan State Academy of Veterinary Medicine named after NE Baumen. 2013. –P. 237 - 241), which is a phytomineral adsorbent of mycotoxins with a probiotic effect. The additive includes mineral sorbents (zeolites, bentonites, silicon oxides, opal), a fermented mixture of mono-, oligosaccharides, pectin, as well as a probiotic component immobilized on phytosorbent (3 strains of Bacillus subtillis, 1 strain of Bacillus licheniformis). The disadvantage of this sorbent is the inclusion of a large amount of amorphous silica (zeolites, opal), which reduce the sorption capacity per unit mass of feed and prevent the binding of mycotoxins.

For the prototype, the Alltech Mikosorb sorbent and the method for its preparation described in US Pat. No. 6,045,834 (published 2000-04-04) were selected. The patent states that the neutralization of such mycotoxins as aflotoxin, zaaraleon, fumonisin, vomitoxin, ochratoxin and T2 toxin is provided due to the synergistic action of mineral clay and biological material, namely, a modified yeast extract of the cell wall of yeast selected from the group consisting of Saccharomycetes, Candida, Kluveromycetes, Torulaspora, or combinations thereof. The effectiveness of the drug Mikosorb proven in vitro experiments.

The composition of the sorbent Mikosorb includes from about 1% to about 10% of mineral clay, consisting of zeolite, bentonite, aluminosilicate or mixtures thereof, and from about 90% to about 99% of the yeast extract of the cell wall.

A method for producing a sorbent comprises modifying the mannanoligosaccharide (MOS) part of the cell wall of the yeast selected from the group consisting of Saccharomycetes, Candida, Kluveromycetes, Torulaspora, or a combination thereof, by exposing the yeast to about 5% to about 20% alcohol during growth, for example during fermentation, which leads to a thickening of the cell wall of the yeast and an increase in the surface area available for binding of mycotoxins in the resulting cell wall extract. According to the invention, any of a number of standard commercially available alcohols can be used, including but not limited to methyl, ethyl and isopropyl alcohols. The obtained yeast cell wall extract is washed several times by centrifugation to remove intracellular components and concentrate the extract. The resulting extract concentrate can be dried by any of a number of methods known in the art, including spray drying or drum drying, to form a hygroscopic, water-soluble powder, which is mixed with powdered clay in the stated ratio, because the physical form of Mikosorb is a dry loose powder suitable for direct inclusion in animal feed or as an additive to the general mixed diet

The disadvantage of the Mikosorb sorbent and the method for its preparation is the need to use a modified yeast extract of the yeast cell wall and the complexity of the method for modifying the yeast wall.

The objective of the group of inventions is to expand the range of sorbents used in the form of a feed additive for the binding of mycotoxins, which can be used in the prevention of diseases of the gastrointestinal tract and intoxications of various etiologies in farm animals and poultry, as well as the development of a method for producing the sorbent in the form of a biocomposite additive.

The technical result of the task is to solve the problem.

The task of developing the composition of the sorbent for the binding of mycotoxins is solved by the proposed biocomposite additives, which include mineral clay and biological materials, which introduced the following new features:

- as mineral clay use sedimentation-enriched and activated with hydrochloric acid 10% montmorillonite-containing clay (hereinafter referred to as MSH) in an amount of from 10 to 20 mass. %, while clay may include illite, quartz, muscovite. Over 20% of MSH is not advisable to use, because it negatively affects the digestive tract of animals, causing constipation;

- fodder yeast, usually obtained in the form of food waste, for example, selected from Pichia pastoris, Faex medicinalis,, Saccaromicete spp, or a combination thereof, in an amount of 80 to 90 wt.% is used as biological material.

A method of obtaining a biocomposite feed additive, comprising mixing mineral clay and biological material for farm animals and poultry contains the following new features:

- Conduct a sedimentation enrichment of montmorillonite containing clay in water, for which they first maintain the suspension for 24 hours, stir up for one minute and again defend the suspension for 20 minutes, then select a supernatant suspension with clay particles less than 10 microns in size from the upper 10-centimeter layer , stand for another 10 minutes and after sedimentation of the suspension, clarified water is decanted, then the precipitate is enriched in MSH in an oven at 70-105 ° C. After the enriched MSH is crushed to a homogeneous consistency;

- enriched MSH is activated with hydrochloric acid 10%, after which it is washed with purified distilled water to a neutral medium, dried at a temperature of not more than 105 ° C and crushed in a ball mill to a particle size of not more than 10 microns;

- biological material in the form of biomass of fodder yeast Pichia pastoris, Faex medicinalis,, Saccaromicete spp, or combinations thereof, is filled with distilled water, mixed and processed in an ultrasonic generator at 1500 Hz (Cryamide, Russia) for 10 minutes 4 times, cooling the system for 5 minutes after each treatment, then centrifuged and the supernatant drained. This treatment gives an increase in the surface area of yeast cells, i.e. increased adsorption capacity of biomass;

- then the remainder of the treated biomass is heated to 40 ° C and MSH is gradually added with constant mechanical stirring in the stated ratio;

- the mixture is thoroughly mixed for at least 40 minutes;

- then subjected to soft drying t = 65 ° C to a moisture content of not more than 30% and grind to a finely divided homogeneous medium.

A comparative analysis of the proposed group of inventions shows that the main difference from the prototype in the claimed composition and method is that the raw material is not modified yeast extract of the yeast cell wall, but the yeast microorganisms Pichia pastoris or Faex medicinalis, or Saccaromicete spp, or combinations thereof pretreated in an ultrasonic generator. Using enriched and activated hydrochloric acid of montmorillonite-containing clay in combination with fodder yeast provides a range of positive effects: minimize the side effect of montmorillonite on the sorption of nutrients in the feed, normalize metabolism in the gastrointestinal tract of animals, increase sorption of mycotoxins, and also increase the introduction of microbiological protein and vitamins due to fodder yeast. Therefore, the claimed group of inventions expands the range of composite sorbents based on MSH, which can be used as feed additives to help neutralize mycotoxins in feed and improve gastrointestinal motility, while not causing allergic, locally irritating, cumulative, mutagenic, teratogenic and carcinogenic effects .

The composition of the proposed biocomposite additives may include, but not necessarily, supplements in the form of amino acids alanine and / or leucine, vitamins, trace elements such as hematite, calcite, essential fatty acids, and other biologically active substances.

The invention is characterized by the following graphic materials:

FIG. 1 Comparative characteristic of the sorption ability to absorb dye methylene blue.

The invention can be illustrated by the following examples of its specific implementation.

Example 1

Montmorillonite containing clay in water is sedimented by soaking the suspension for 24 hours, agitate for one minute and sediment the suspension for 20 minutes, then add a supernatant suspension with a clay particle size of less than 10 microns from the upper 10-centimeter layer, assert another 10 minutes and after sedimentation of the suspension, clarified water is decanted, then the precipitate is enriched in MSH in an oven at 70-105 ° C. With the appropriate equipment, the drying process can be carried out at temperatures up to 900 ° C, which reduces the duration of the process, but increases its energy intensity. After the enriched MSH is crushed to a homogeneous consistency. Then MSH is activated with hydrochloric acid 10%, in a ratio of 1 part clay to 2 parts acid at room temperature until gas bubbles cease to exist, then it is washed with purified distilled water to a neutral medium, dried at a temperature of not more than 105 ° C and crushed in a ball mill to particle sizes of not more than 10 microns.

Example 2

Obtaining a biocomposition with a content of 20% MSH

500g Pichia pastoris yeast biomass is poured with distilled water of 100-300 ml depending on the degree of compaction of the biomass and mixed to a homogeneous mass with a magnetic stirrer for 15 minutes. Then it is processed in an ultrasonic generator at 1500 Hz (Cryamide, Russia) for 10 minutes. 4 times, after each treatment, cool the system for 5 minutes The treated biomass is centrifuged at 15,000 rpm, the supernatant is drained, the biomass precipitate is heated to 40 ° C, and 125 g of enriched MSH are added with constant mechanical stirring. The mixture is thoroughly mixed for at least 40 minutes and subjected to soft drying t = 65 ° C to a moisture content of not more than 30%. Then crushed in a ball mill to a particle size of not more than 10 microns

Example 3

Obtaining a biocomposition with a content of 15% MSH.

Faex medicinalis yeast biomass 500 g is poured with distilled water 100 - 300 ml depending on the degree of compaction of the biomass and mixed until smooth with a magnetic stirrer for 15 minutes. Then it is processed in an ultrasonic generator at 1500 Hz (Cryamide, Russia) for 10 minutes. 4 times, after each treatment, cool the system for 5 minutes The treated biomass is centrifuged at 15,000 rpm, the supernatant is drained, the biomass precipitate is heated to 40 ° C, and 88 g of enriched MSH are added with constant mechanical stirring. The mixture is thoroughly mixed for at least 40 minutes and subjected to soft drying t = 65 ° C to a moisture content of not more than 30%. Then crushed in a ball mill to a particle size of not more than 10 microns

Example 4

Obtaining a biocomposition with a content of 10% MSH.

Saccaromicete spp yeast biomass 500 g is poured with distilled water 100 - 300 ml depending on the degree of compaction of the biomass and mixed until smooth with a magnetic stirrer for 15 minutes. Then it is processed in an ultrasonic generator at 1500 Hz (Cryamide, Russia) for 10 minutes. 4 times, after each treatment, cool the system for 5 minutes The treated biomass is centrifuged at 15,000 rpm, the supernatant is drained, the biomass precipitate is heated to 40 ° C and 55.5 g of enriched MSH are added with constant mechanical stirring. The mixture is thoroughly mixed for at least 40 minutes and subjected to soft drying t = 65 ° C to a moisture content of not more than 30%. Then crushed in a ball mill to a particle size of not more than 10 microns

Example 5

The study of the adsorption efficiency of the biocomposition prepared according to methods 2, 3 and 4, as well as the Mycosorb preparation according to the prototype, was carried out according to the method described in GOST 4453-74. Figure 1 presents a comparative characteristic of the adsorption capacity of the claimed composition for the absorption of the dye methylene blue. It can be seen that the biocomposition prepared according to examples 2, 3, and 4 has an adsorption capacity for dye methylene blue almost two times greater than that of the prototype.

Example 6

To determine the sorption ability of the proposed biocomposite, the standard method was used - the In Vitro Method described in the source (Malkov M.A., Bogomolov V.V., Dankova T.V., Krasnov K.A. Mycotoxins - a strategy for eliminating their effect on the body farm animals and poultry // Tsenovik. 2012. - No. 1 (181) - S.74–89). According to this method, based on the difference between the amount of adsorption of toxins during the stay in the stomach and the amount of desorption of the toxin released from the sorbent during the residence of the feed in the intestine, the practical coefficient of performance (PCPD), which is a criterion for assessing the quality of mycosorbent, is determined. PCAP has a percentage. The higher it (Net Efficiency), the more effective the adsorption, and therefore the amount of deactivated and bound mycotoxin increases. The sorption of mycotoxins is quantified by changing the pH, which allows you to simulate a change in the acidity of the medium in the digestive tract of animals.

For research, mycotoxins fumonisin, T-2 toxin and zearalenone were obtained in laboratory conditions by direct contamination under aseptic conditions of sunflower and soybean meal with toxigenic strains of mold fungi of the genera Aspergillus ochraceus, Penicillium viridicatum and Fusarium gramineum extraction using methods of extraction IFA.

The results of the determination of PCPD in relation to the sorption efficiency of the claimed biocomposite obtained in examples 2, 3, 4 in comparison with native clay and the prototype are shown in table 1

Tab. 1.

Mycotoxin PKPD,
% The biocomposition obtained in Example 2 Fumanisin 89.8 T-2 Toxin 7.4 Zearalenone 15,5 The biocomposition obtained in Example 3 Fumanisin 90.3 T-2 Toxin 7.2 Zearalenone 15.7 The biocomposition obtained in Example 4 Fumanisin 90.5 T-2 Toxin 7.3 Zearalenone 15.4 Native MSH Fumanisin 74.9 T-2 Toxin 2.0 Zearalenone 5,0 Mycosorb Fumanisin 14,4 T-2 Toxin 7.3 Zearalenone 0

The results of the study, shown in table 1, showed that the PCPD with respect to the sorption efficiency of the claimed biocomposition obtained in examples 2, 3 and 4, is much higher in comparison with Mycosorb and native clay.

Thus, the task to expand the range of sorbents and to develop a biocomposition for use in the form of a feed additive for the binding of mycotoxins, which can be used in the prevention of diseases of the gastrointestinal tract and intoxications of various etiologies in farm animals and poultry, as well as the method for its preparation, is achieved .

Claims (4)

1. The composition of the biocomposite feed additives for farm animals and poultry, including mineral clay and biological materials, characterized in that the mineral clay is enriched and activated with hydrochloric 10% acid montmorillonite-containing clay, and the biomass of fodder yeast selected from biological material is selected Pichia pastoris, Faex medicinalis, Saccaromicete spp or combinations thereof, in the following ratio of starting components, wt.%: enriched and activated hydrochloric 10%  montmorillonite-containing clay from 10 to 20 fodder yeast biomass from 90 to 80 2. The method for producing a biocomposite feed additive for farm animals and poultry according to claim 1, characterized in that the montmorillonite-containing clay is preliminarily enriched and activated with hydrochloric acid 10%, then washed with purified distilled water to a neutral medium, dried at a temperature of not more than 105 ° C and crushed to a homogeneous consistency, and biological material in the form of biomass of fodder yeast selected from Pichia pastoris, Faex medicinalis, Saccaromicete spp, or a combination thereof, is poured with distilled water before mixing, mixed to a homogeneous mass and processed in an ultrasonic generator at 1500 Hz 4 times 10 minutes, cooling the system for 5 minutes after each treatment, then the treated yeast biomass is centrifuged, the supernatant is drained, the biomass sediment is heated to 40 ° C and enriched montmorillonite-containing clay is gradually added with constant mechanical stirring, the mixture is thoroughly mixed for at least 40 minutes, then subjected to soft drying at t = 65 ° C to a moisture content of not more than 30% and grind to a finely divided homogeneous medium. 3. The method for producing a biocomposite feed additive according to claim 2, characterized in that the enriched montmorillonite-containing clay is activated with hydrochloric acid 10%, in a ratio of 1 part clay to 2 parts acid at room temperature until gas bubbles cease to be released.

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