RU2528747C1 - Method of production of preparation for prevention of digestive tract infections in agricultural poultry and preparation obtained by method - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: method of production of preparation for prevention of digestive tract infections in agricultural poultry comprises preliminary mixing the yeast biomass of Saccharomyces cerevisiae with enzymatic complexes "CelloLux-A" and "Protosubtilin", lactic acid, fumaric acid, and water. The ratio of the components in the mixture is: enzymatic complex "CelloLux-A" 0.2-10 wt %, enzymatic complex "Protosubtilin" 0.2-10 wt %, lactic acid 0.5-10 wt %, fumaric acid 0.5-10 wt %, water 0.5-10 wt %, yeast biomass is the rest. The resulting mixture is subjected to mechanical activation in the jet type activators providing acceleration of the processed material of 30-450 m/s² and the residence time in the processing zone of 0.25-15 min. Also a preparation obtained by this method is provided.

EFFECT: group of inventions provides increase in productivity of agricultural poultry, promotes gross growth, has a complex action on pathogenic and beneficial microflora of the digestive tract, stimulating the growth of beneficial microflora.

2 cl, 4 tbl, 5 ex

Description

The invention relates to veterinary medicine, which makes it possible to obtain a preparation from yeast biomass Saccharomyces cerevisiae whose active ingredients are mannoproteins integrated into a partially hydrolyzed yeast cell wall, water-soluble mannoproteins, lactic and fumaric acids. The drug can be used as a prebiotic and for the prevention of digestive tract infections in animal husbandry and poultry farming.

In animals, it is known that there is an important relationship between digestion, nutrient availability, and the bacteria that inhabit the intestinal tract. The combination of these factors affects not only the amount of available nutrients necessary for growth and maintaining the health of the body, but also for the prevention of diseases, especially in relation to the introduction of pathogenic microorganisms.

Adding antibiotics to the food for therapeutic or prophylactic purposes reduces the total number of bacteria in the digestive tract, which negatively affects health. Thus, the urgent task is to create environmentally friendly drugs with a prebiotic effect, capable of adjusting the content of harmful and beneficial microflora in the digestive tract.

Potentially suitable prebiotics include preparations containing acidifiers - fumaric, lactic, citric, succinic and malic acids. Due to the introduction of these substances into the diet of animals, the environment in the gastrointestinal tract (GIT) becomes more acidic, which stimulates beneficial microflora and indirectly reduces the number of pathogenic microorganisms.

The next class of drugs that correct the microflora of the gastrointestinal tract by removing pathogenic microorganisms from it is mannanoligosaccharide and mannoprotein preparations. Sorbed on specific surface areas of pathogens, this class of compounds prevents the attachment of pathogens to the walls of the gastrointestinal tract and helps to remove them from the body. Mannanoligosaccharide and mannoprotein preparations are known to act on most varieties of E. coli. Salmonella and Cryptosporidium, without removing the beneficial microflora from the body.

The invention is known (1. RU No. 2359465 C1, publ. 06/27/2009), the essence of which is that for feeding young pigs use dry beet pulp, additionally containing probiotic, which includes bacterial cultures of Bifidobacterium globosum, Enterococcus faecium , Bacillus subtilis, fumaric acid and zeolites. This feed additive allows you to increase consumption and improve the use of young pigs feed containing dry beet pulp, which helps to increase the average daily increase in live weight of young pigs and reduce the cost of production of pig production.

However, the present invention has a number of disadvantages, for example, the use of bacterial cultures that require special conditions for the production, storage and transportation. In addition, this composition is only able to indirectly affect the amount of harmful microflora, reducing it only by improving the conditions for beneficial microorganisms.

On the other hand, in the invention (2. USA No. 7048937 B2, published May 23, 2006), the authors propose to use a preparation containing mannoproteins integrated into a partially hydrolyzed cell wall as a means of coccidiosis. In this case, the drug has a rather low antibacterial activity due to the fact that the process of its preparation involves discarding the water-soluble fraction containing water-soluble mannoproteins. Also, the invention does not solve the issue of stimulating beneficial microflora and it is believed that the removal of pathogens from the body indirectly contributes to this.

The closest technical solution selected for the prototype is the invention (3. Pat. RU No. 2406516, published 02.12.2010). According to the invention, the composition of the antibacterial preparation from the biomass of yeast Saccharomyces cerevisiae includes mannoproteins integrated into a partially hydrolyzed cell wall, water-soluble mannoproteins, water-soluble proteins and polysaccharides, as well as an enzymatic complex with depolymerase activity to β-glucan. A method of obtaining a preparation according to the prototype includes mixing yeast biomass with an enzymatic complex, mechanically activating the mixture, obtaining a tablet form of the product, followed by heating at the temperature of the enzymes.

The method of obtaining the drug according to the prototype has disadvantages. The mixture of components according to the prototype cannot be efficiently processed in jet mills, and the equipment proposed by the authors operates in inefficient operating modes. This imposes the need for additional energy-intensive stages of obtaining tabletted forms and heating for enzymatic hydrolysis. The implementation of the production of the drug according to the prototype in this regard is laborious, contains a large number of stages and is economically unjustified.

The composition of the drug according to the prototype also has several disadvantages. So, this composition has insufficient biological activity due to the fact that its mannoproteins are capable of excreting pathogenic microorganisms, but do not directly contribute to the growth and reproduction of beneficial microflora. The use of an enzymatic complex having depolymerase activity with respect to β-glucan as a hydrolyzing agent allows the hydrolysis of yeast glucan without affecting the protein components of biomass. This fact significantly reduces the efficiency of enzymatic hydrolysis and negatively affects the technological and economic component of the production method.

The problem solved by the claimed technical solution is to create a drug with a complex effect on the pathogenic and beneficial microflora of the gastrointestinal tract (GIT) of farm animals and poultry, as well as the development of an environmentally friendly, simple way to obtain this drug, allowing the use of available raw materials, such like fodder yeast biomass, post-alcohol stillage, etc.

The problem is solved due to the fact that the method of obtaining the drug for the prevention of digestive tract infections in poultry includes pre-mixing the yeast biomass Saccharomyces cerevisiae with the enzymatic complexes CelloLux-A and Protosubtilin, lactic acid, fumaric acid and water, mechanical activation of the resulting mixture in the activators of the jet type, providing acceleration of the processed material 30-450 m / s ² and a residence time in the treatment area of 0.25-15 minutes, to obtain the drug, in the following ratio of components in the mixture (wt.%):

- enzymatic complex "CelloLux-A" - 0.2-10;

- enzymatic complex "Protosubtilin" - 0.2-10;

- lactic acid - 0.5-10;

- fumaric acid - 0.5-10;

- water - 0.5-10;

- yeast biomass - the rest.

Significant differences of the claimed drug is its composition.

The drug obtained in this way can be used to prevent infections of the digestive tract in poultry.

The technical result consists in creating a preparation having a complex effect on the pathogenic and beneficial microflora of the gastrointestinal tract of farm animals and poultry and exhibiting sorbent properties due to the presence of mannoproteins, as well as a prebiotic that stimulates the growth of beneficial intestinal microflora.

It is worth noting that with the combined action of mannoproteins (on pathogenic bacteria) and used acids (on beneficial bacteria), the observed biological effect is significantly higher than with the separate use or use of the usual mixture of these components. This is due to the fact that in the mass ratios claimed in this invention, a synergistic effect is observed, consisting in the mutual strengthening of the action of the individual components. The proposed composition cannot be obtained by routine enumeration of reagents.

During mechanical activation of a mixture of yeast biomass with the declared enzymatic complexes and acids, partial mechanical destruction of the cells occurs, as well as disordering of the supramolecular structure of the cell walls and partial hydrolysis of the polymers of the cell wall, which does not occur in the prototype method.

Due to the partial hydrolysis of the polymers of the cell wall during mechanical activation, there is no need for the stages of tabletting and heating according to the prototype, and it is also possible to use more efficient operating modes of mechanochemical equipment. When implementing the inventive method, the release of the target substances into a bioavailable form is facilitated, and the yield is greater than that of the prototype.

The selection of equipment that distinguishes the invention from the prototype cannot be carried out without a comprehensive study of the processes taking place in the cell walls. The conditions for mechanical activation cannot be established by a routine enumeration of modes and are not obvious.

The patent search did not reveal drugs of a similar composition, therefore, it was concluded that the claimed technical solution meets the criterion of "novelty." The set of essential features of the proposed method is also not identified, which allows us to conclude that the claimed drug and its production method meet the criterion of "inventive step".

The invention is illustrated by the following examples.

Example 1

A mixture is prepared according to Table 1, containing Saccharomyces cerevisiae yeast biomass, CelloLux-A enzyme complex (manufactured by PO SIBBIOFARM LLC Berdsk), which has depolymerase activity with respect to β-glucan, Protosubtilin enzyme complex (manufactured by LLC PO SIBBIOFARM Berdsk), which has depolymerase activity with respect to protein molecules, lactic and fumaric acids, water.

Table 1 The composition of the mixture prepared for mechanical activation Mixture component The content of the component, wt.% Saccharomyces cerevisiae yeast biomass 93.6 Enzymatic complex "CelloLux-A" 0.2 Enzymatic complex "Protosubtilin" 0.2 Lactic acid 0.5 Fumaric acid 0.5 Water 5

The mixture is mechanically activated in the activators of the jet type, providing an acceleration of the processed material of 30 m / s and a residence time in the processing zone of 0.25 minutes.

When the content of enzymatic complexes and acids is below 0.2% and 0.5%, respectively, the hydrolysis reaction proceeds slightly, which negatively affects the composition of the drug. When accelerating the processed material of less than 30 m / s ² , the introduction of particles of enzymatic complexes and acids into the particles of yeast biomass is not ensured, which also reduces the efficiency of hydrolysis. The use of processing less than 0.25 minutes does not provide the necessary accumulation of free energy in the form of deformations of the cell walls and is not practical from an economic point of view.

The resulting product contains biologically active components in the following ratio, wt.%:

- mannoproteins integrated into the cell wall of yeast - 1.3;

- water-soluble mannoproteins of the yeast cell wall - 0.3;

- water-soluble proteins of yeast biomass - 5;

- water-soluble glucosaccharides of yeast biomass - 4,

- enzymatic complex with depolymerase activity with respect to β-glucan - 0.2;

- enzymatic complex with depolymerase activity in relation to protein molecules - 0.2;

- lactic acid - 0.5;

- fumaric acid - 0.5;

- water - 5;

- the remainder of the mechanically activated biomass of yeast - the rest.

Example 2

A mixture is prepared according to Table 2, containing Saccharomyces cerevisiae yeast biomass, the enzyme complex CelloLux-A (manufactured by PO SIBBIOFARM LLC in Berdsk), which has depolymerase activity with respect to β-glucan, the Protosubtilin enzyme complex (manufactured by LLC PO SIBBIOFARM Berdsk), which has depolymerase activity with respect to protein molecules, lactic and fumaric acids, water.

table 2 The composition of the mixture prepared for mechanical activation Mixture component The content of the component, wt.% Saccharomyces cerevisiae yeast biomass 75 Enzymatic complex "CelloLux-A" 3 Enzymatic complex "Protosubtilin" 3 Lactic acid 7 Fumaric acid 7 Water 5

The mixture is mechanically activated in the activators of the jet type, providing acceleration of the processed material 450 m / s and a residence time in the treatment zone of 15 minutes.

The content of enzymatic complexes and acids above 3% and 7%, respectively, does not increase the depth of the hydrolysis of yeast biomass and is not economically feasible. When the processed material is accelerated more than 450 m / s ² for more than 15 minutes, partial denaturation of the target mannoproteins is observed.

The resulting product contains biologically active components in the following ratio, wt.%:

- mannoproteins integrated into the cell wall of yeast - 4.7;

- water-soluble mannoproteins of the yeast cell wall - 3.0;

- water-soluble proteins of yeast biomass - 12;

- water-soluble glucosaccharides of yeast biomass - 17;

- enzymatic complex with depolymerase activity in relation to β-glucan - 3;

- enzymatic complex with depolymerase activity in relation to protein molecules - 3;

- lactic acid - 7;

- fumaric acid - 7;

- water - 5;

- the remainder of the mechanically activated biomass of yeast - the rest.

Example 3

A mixture is prepared according to Table 3, containing yeast post-alcohol distillery stillage (TU 9296-302-00008064-98), the enzyme complex CelloLux-A (manufactured by PO SIBBIOFARM LLC in Berdsk), which has depolymerase activity with respect to β-glucan , Protosubtilin enzyme complex (manufactured by LLC SIBBIOFARM Berdsk), which has depolymerase activity with respect to protein molecules, lactic and fumaric acids, water.

Table 3 The composition of the mixture prepared for mechanical activation Mixture component The content of the component, wt.% Post-alcohol Bard 62 Enzymatic complex "CelloLux-A" 10 Enzymatic complex "Protosubtilin" 10 Lactic acid four Fumaric acid four Water 10

The mixture is mechanically activated under the conditions of Example 2.

In the hydrolysis of yeast post-alcohol stillage stillage, the optimal content of enzymatic complexes and acids is 10% and 4%, respectively. It is also necessary to use the most energy-intensive conditions of mechanical activation, due to the difficulty of extracting target substances from the yeast cells contained in the bard.

The resulting product contains biologically active components in the following ratio, wt.%:

- mannoproteins integrated into the cell wall of the yeast - 1.8;

- water-soluble mannoproteins of the yeast cell wall - 0.7;

- water-soluble proteins of yeast biomass - 16;

- water-soluble glucosaccharides of yeast biomass - 25;

- enzymatic complex with depolymerase activity in relation to β-glucan - 10;

- enzymatic complex with depolymerase activity in relation to protein molecules - 10;

- lactic acid - 4;

- fumaric acid - 4;

- water - 10;

- the remainder of the mechanically activated biomass of yeast - the rest.

Involving in the production of cheap and substandard raw materials allows you to rationally dispose of the listed waste, as well as reduce the cost of obtaining the drug.

Example 4

A mixture is prepared according to Table 4, containing biomass of fodder yeast (GOST 80083-74 or TU 9291-003-12914410-03), the enzyme complex "CelloLux-A" (manufactured by LLC SIBBIOFARM Berdsk), which has depolymerase activity of with respect to β-glucan, the Protosubtilin enzymatic complex (manufactured by PO SIBBIOFARM LLC in Berdsk), which has depolymerase activity with respect to protein molecules, lactic and fumaric acids, water.

Table 4 The composition of the mixture prepared for mechanical activation Mixture component The content of the component, wt.% Fodder Yeast Biomass 69.5 Enzymatic complex "CelloLux-A" 5 Enzymatic complex "Protosubtilin" 5 Lactic acid 10 Fumaric acid 10 Water 0.5

The mixture is mechanically activated in the activators of the jet type, providing acceleration of the processed material 370 m / s ² and a residence time in the treatment zone of 12 minutes.

In the case of processing biomass of fodder yeast, the optimal content of enzymatic complexes and acids is 5% and 10%, respectively. The optimal modes of mechanical activation is the acceleration of the processed material 370 m / s and 12 minutes, respectively.

The resulting product contains biologically active components in the following ratio, wt.%:

- mannoproteins integrated into the cell wall of yeast - 2.5;

- water-soluble mannoproteins of the yeast cell wall - 1.1;

- water soluble proteins of yeast biomass - 45;

- water-soluble glucosaccharides of yeast biomass - 11;

- enzymatic complex with depolymerase activity in relation to β-glucan - 5;

- enzymatic complex with depolymerase activity in relation to protein molecules - 5;

- lactic acid - 10;

- fumaric acid - 10;

- water - 0.5;

- the remainder of the mechanically activated biomass of yeast - the rest.

Example 5

It is carried out in the conditions of example 2. The work was carried out in the vivarium of the FSUE Zagorsk EPH VNITIP of the Russian Agricultural Academy (Report dated March 01, 2013). The resulting product in an amount of 0.1 wt.% Was added to the diet of broilers cross "Cobb 500" from November 13 to December 21, 2012. Animals of the control group in the diet received the product.

The gross increase in experimental animals was 2.5% higher than in animals from the control group. The experimental animals spent 2.3% less feed per 1 g of live weight gain. The average daily gain was 2.7% higher than in the control group.

The increase in broiler productivity was due to improved digestibility and use of feed nutrients in the experimental group. So, digestibility of dry matter of feed was higher than in the control group by 2.7%.

The difference in protein digestibility was 1.3%, fat - by 2.0%, fiber - 1.8%. Feed nitrogen utilization was 1.9% higher than control.

The chemical composition of meat, blood and internal organs was within normal limits and did not differ from the composition of samples taken in the control group.

Microbiological control of the microflora of feces and samples of the gastrointestinal tract revealed pathogens in the control group that were not found in the samples of the experimental group.

Histological studies showed that the histoarchitectonics of the small intestine in animals of the control group is impaired. The general morphofunctional state of organs reflects the developing processes in the form of protein dystrophy and pronounced desquamation of enterocytes of the small intestine, accompanied by hemocirculatory disorders.

At the same time, the histoarchitectonics of the small intestine of experimental animals has been preserved and is characteristic of healthy broiler chickens in this period of development.

Thus, the claimed drug has a high and complex biological activity, has been tested and therefore meets the criterion of "industrial applicability".

Claims (2)

1. A method of obtaining a preparation for the prevention of digestive tract infections in poultry, comprising pre-mixing the yeast biomass Saccharomyces cerevisiae with the enzyme complexes CelloLux-A and Protosubtilin, lactic acid, fumaric acid and water, mechanical activation of the resulting mixture in activators of jet type providing acceleration of the processed material 30-450 m / s ² and a residence time in the treatment zone of 0.25-15 minutes, with the preparation, in the following ratio of components in the mixture and (wt.%):
- enzymatic complex "CelloLux-A" - 0.2-10;
- enzymatic complex "Protosubtilin" - 0.2-10;
- lactic acid - 0.5-10;
- fumaric acid - 0.5-10;
- water - 0.5-10;
- yeast biomass - the rest. 2. The drug for the prevention of digestive tract infections in farm birds, obtained by the method according to claim 1.