BG113040A - Probiotic for animal husbandry, preparation method and application thereof - Google Patents

Abstract

The invention relates to a probiotic for animal husbandry, containing a microbial complex consisting of Lastobacillus acidophilus NBIMCC 8242, Lactobacillus helveticus NBIMCC 8269, Lactobacillus delbrueckii subsp. bulgaricus NBIMCC 8244, Lactobacillus delbrueckii subsp. lactis NBIMCC 8250, Streptococcus thermophilus NBIMCC 8253, Enterococcus faecium NBIMCC 8270 in an amount of 1 x 10 9 cfu / g and plant substrate derived from sunflower seed flour containing crude protein from 25.0 to 30.0% by weight, carbohydrates from 8.0 up to 11.0%, fat from 0.3 to 0.6%, fiber from 8.0 to 15.0% and free amino acids from 39.0 to 40.0%, such as lysine, methionine, cysteine, threonine, leucine , methylcysteine, aspartic acid, serine, glutamic acid, glycine, alanine, valine, isoleucine, phenylalanine, histidine, arginine. The invention also relates to a method for preparing the probiotic, consisting of dissolving sunflower seed flour in water, enzymatic treatment with a proteolytic enzyme selected from the group of serine proteases and a cellulotic enzyme selected from cellulase, phytase, after cooling the mixture what follows is fermentation with starter culture of Lactobacillus acidophilus NBIMCC 8242, Lactobacillus helveticus NBIMCC 8269, Lastoballus delbrueckii subsp. bulgaricus NBIMCC 8244, Lactobacillus delbrueckii subsp. lactis NBIMCC 8250, Streptococcus thermophilus NBIMCC 8253, Enterococcus faecium NBIMCC 8270 and carrying out a fermentation process during which is applied neutralization of the organic acids accumulating during the process, and afterwards the enriched product is cooled and subjected to pulverization. The application of the probiotic in animal husbandry leads to an increase in live weight of the animals while providing healthy nutrition and breeding, suppressing the putrefactive processes in the intestine and helping to restore and maintain beneficial gut microflora, further having a beneficial effect on intestinal function. All bacterial strains involved are isolated from environmentally friendly natural sources and are not genetically modified. The method, according to the invention, provides a product with a high content of beneficial microorganisms, produces metabolic substrates, such as vitamins and short-chain fatty acids, increases the content of soluble peptides, which leads to the production of natural antioxidants, and not least improves the functional properties of the product.

Description

BACKGROUND OF THE INVENTION

Patent BG 65 466 discloses a probiotic which is used as an additive in the feed or water of farm animals, in particular in the industrial rearing of poultry, pigs, bovine animals and small animals, in order to provide a regulatory effect on the gastrointestinal tract. , suppression or complete destruction of pathogenic microorganisms, ensuring the availability of a wide range of enzyme complexes, resulting in increased animal weight. The probiotic is a mixture of lyophilized monocultures of eight strains of lactic acid microorganisms: Lactobacillus acidophilus, NBPMKK 8242; Lactobacillus helveticus, N NBPMCC 8268; Lactobacillus delbrueckii subsp. Lactis, N NBPMCC 8269; Lactobacillus delbrueckii subsp. bulgaricus, N NBPMCC 8250; Lactobacillus delbrueckii subsp. bulgaricus, N NBPMCC 8243; Lactobacillus delbrueckii subsp. bulgaricus, N NBPMCC 8247; Streptococcus thermophilus, N NBPMKK 8253Streptococcus faecium (Enterococcus faecium), NNBPMKK8270.

The method of preparation consists in the following: each of the strains is cultured separately in a suitable nutrient medium, and after the development of the particular monoculture reaches its logarithmic phase, 25% ammonia water or 20% sodium hydroxide are gradually added, while in the finished product remains 1.5-2% free lactic acid, after completion of the fermentation process, the cultures are cooled to 10 ° C, subjected to lyophilization and mixed in the desired proportions.

Use by adding different amounts of the individual crops to granulated feed for chickens, to compound feed for piglets, to compound feed for chickens is described.

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A product is known - a probiotic for animal husbandry (product specification), consisting of the following microorganism cultures, in lyophilized form: Lactobacillus acidophilus NBIMCC 8242; Lactobacillus helveticus NBIMCC 8269; Lactobacillus delbrueckii subsp. bulgaricus NBIMCC 8244; Lactobacillus delbrueckii subsp. lactis NBIMCC 8250; Streptococcus thermophilus NBIMCC 8253; Enterococcus faecium NBIMCC 8270

Known probiotics have unsatisfactory survival and multiplication when ingested by animals, when mixed with feed, for different types of pets. The production method involves the use of special nutrient media and separate cultivation of monocultures.

RU 2 546 880 C2 describes a method for preparing a probiotic preparation for farm animals, which consists in the following: a phytonutrient is prepared by mixing 404.5 kg of cellulose-containing vegetable substrate with tap water, 150 160 g of baker's yeast autolysate , 2 - 3 g of potassium phosphate, 0,125 - 0,130 g of magnesium sulphate, 1,5 - 1.8 g of molasses and after sterilization, a culture of Cellulomonas flavigena VKPMV-2559 is introduced into the obtained phytocarrier and cultured at 45 - 47 ° C for 4

- 5 hours, then liquid cultures of Bacillus subtilis VKPMV-8130, Lactococcus lactis subsp.lactis Fl 16 VKPMB4591 are introduced in equal amounts, the resulting mixture is stirred and left at 37 - 38 ° C for 48 - 50 hours and dried to humidity 8 -

%. Monocultures are cultivated on special nutrient media. Ground straw, sunflower or soybean meal is used as a phytonutrient.

According to the described method, a final product is obtained, containing live cells of three v \ <3M <r.Bacillus subtilis VKPM B-8130, Lactococcus lactis subsp.lactis F-116 VKPM B4591, Cellulomonas flavigena VKPM B-2559, products of the metabolism of microorganisms and products of the biotransformation of cellulose fitonositel onto which a ^cultured micro-organisms by incomplete solid-state fermentation. The probiotic is in the form of a coating on the cellulose carrier.

This method is complicated, time consuming, too energy consuming. The composition of the final product is unstable, variable, difficult to control and the effect of its application has an unsatisfactory spectrum.

The aim of the present invention is to provide a probiotic for animals, which contains a probiotic with a high concentration, which leads to an increase in live weight of animals while providing healthy nutrition and breeding, suppressing putrefactive processes in the intestine and helping to restore and maintain beneficial intestinal microflora, to have a beneficial effect on intestinal function through the production of lactic acid, to have pronounced antimicrobial activity against pathogenic microorganisms such as E. coli, Listeria ssp., St.aureus, which leads to a reduction in morbidity and mortality.

In particular, the aim of the present utility model is to create a probiotic consisting of lactic acid bacteria, which provides improved viability of microorganisms and multiplication, to achieve higher indicators of its properties such as high antimicrobial activity, better adhesion to intestinal surface and high bioavailability of cells, better digestibility of feed.

The aim is also to improve the method of obtaining, in the direction of reducing energy costs, duration of processes. In addition, through the method to fully utilize the waste product.

SUMMARY OF THE INVENTION

According to the present invention there is provided a probiotic for animals comprising a microbial complex comprising Lactobacillus acidophilus NBIMCC 8242, Lactobacillus helveticus NBIMCC 8269, Lactobacillus delbrueckii subsp. bulgaricus NBIMCC 8244, Lactobacillus delbrueckii subsp. lactis NBIMCC 8250, Streptococcus thermophilus NBIMCC 8253, Enterococcus faecium NBIMCC 8270 in an amount of Ix10 ⁹ cfu / g and vegetable substrate of sunflower meal containing crude protein from 25.0 to 30.0% by weight, carbohydrates from 8.0 to 11.0 %, fat - 0.3 - 0.6%, fiber from 8.0 to 15 and free amino acids 39.0 to 40.0%, such as lysine, methionine, cystine, threonine, leucine, methylcystine, aspartic acid, serine, glutamic acid, glycine, alanine, valium, isoleucine, phenylalanine, histidine, arginine.

The process for preparing the probiotic according to the invention consists in the following: sunflower flour is dissolved in water to a dry matter of 8 - 12%, and at temperatures of 45 - 52 ° C and pH 6-9 an enzymatic treatment with a proteolytic enzyme selected from the group of serine proteases, in an amount of 0.05 to 0.15% by volume of the substrate and celluloid: _{e <} 4 ·; ·.:. · * Enzyme selected from cellulase, phytase in an amount of 0.05 to 0.15%. After cooling the mixture to 37-40 ° C, it is fermented with a starter culture in an amount of 0.05 to 0.1%, and a fermentation process is carried out for 6 to 20 hours, during which neutralization of organic acids is applied. accumulating during the process. Then, the enriched product is cooled and subjected to spray drying at a temperature drop from 185 - 190 ° C to 90 ° C to a dry matter content of 94 - 97%.

The current probiotic, consisting of lactic acid bacteria, a product of plant origin - sunflower meal, has improved survival, viability of microorganisms and very good reproduction, has high antimicrobial activity against pathogenic microorganisms such as E. coli, Listeria ssp. aureus, which reduces morbidity and mortality, has better adhesion to the intestinal surface and shows high cell bioavailability. The protein product from sunflower flour is a new product, developed especially for inclusion in a probiotic composition, where it functions as a prebiotic and its constituent substances are used to the maximum. The composition has a high protein content, increased the amount of free amino acids, oligosaccharides, which leads to easier digestibility in animal nutrition. In addition to these components, it also contains metabolites of bacterial cultures, unchanged cellulose, trace elements, calcium, phosphorus, vitamins, folic acid. Its use in animal husbandry leads to an increase in live weight of animals while providing healthy nutrition and breeding, suppressing putrefactive processes in the intestine and helping to restore and maintain beneficial intestinal microflora, has a beneficial effect on intestinal function through the production of lactic acid.

The method according to the invention provides a product with a high content of beneficial microorganisms, produces metabolic substrates, such as vitamins and short-chain fatty acids, increases the content of soluble peptides, which leads to the production of natural antioxidants, improves the functional properties of the product. The result of the method is a high biological, economic and ecological effect.

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EXAMPLE PERFORMANCE

The process for preparing the probiotic according to the invention consists in the following: sunflower flour is dissolved in water to a dry matter content of 10%, and at temperatures of 50 ° C and pH 8 an enzymatic treatment with 0.1% proteolytic enzyme alkalase and cellulase is carried out. After cooling the mixture to 37 ° C, fermentation with a starter culture of Lactobacillus acidophilus NBIMCC 8242, Lactobacillus helveticus, WlCC 8269, Lactobacillus delbrueckii subsp. bulgaricus NBIMCC 8244, Lactobacillus delbrueckii subsp. lactis NBIMCC 8250, Streptococcus thermophilus NBIMCC 8253, Enterococcus faecium NBIMCC 8270 in an amount of 0.08%, and carrying out a fermentation process for 10 hours, during which neutralization of organic acids accumulating during the process with ammonia water is applied . The pH is maintained at 5.6. Then, the enriched product is cooled and subjected to spray drying at a temperature drop from 190 ° C to 90 C to a dry matter content of 97%.

Proteolytic serine proteases and cellulolytic enzymes such as alkalase and phytase are used in the method. Alkalase in the presence of water (with specific serine endopeptidase) is responsible for stereo-selective hydrolysis of amino esters and selective esters, but also for protein hydrolysis and transesterification and transpeptidation. When using sunflower meal as a source of nutrients, the main problem is the insufficient degree of hydrolysis, respectively, low content of free amino acids and short-chain peptides. Its degree of hydrolysis is decisive for the efficiency of fermentation of probiotic strains and the assimilation of feed. Fermentation with hydrolyzed sunflower flour and lactic acid bacteria offers a number of advantages, especially for the industrial implementation of this process.

The use of cellulolytic enzyme has a very good biological, economic and environmental effect, increasing the absorption of phosphorus phytate from feed and the rate of absorption of calcium, zinc, copper, magnesium, proteins, amino acids, starch and lipids from feed. At the same time, the phosphorus content in animal excrement and environmental pollution is reduced.

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Example of a probiotic:

The obtained probiotic contains Lactobacillus acidophilus NBIMCC 8242, Lactobacillus helveticus NBIMCC 8269, Lactobacillus delbrueckii subsp. bulgaricus NBIMCC 8244, Lactobacillus delbrueckii subsp. lactis NBIMCC 8250, Streptococcus thermophilus NBIMCC 8253, Enterococcus faecium NBIMCC 8270 in an amount of 1 χ 10 ⁹ cfu / g and vegetable substrate of sunflower flour containing crude protein of 30.0% by weight, carbohydrates - 11%, fat - 0.6% , fiber 15.0%, and free amino acids 40.0%, such as lysine, methionine, cystine, threonine, leucine, methylcystin, aspartic acid, serine, glutamic acid, glycine, alanine, valine, isoleucine, phenylalanine, histidine, arginine.

The bacterial strains included in the probiotic complex have been selected for their specific beneficial effects on animal health. They survive and multiply successfully in the animal's gastrointestinal tract and have an inhibitory effect on pathogenic microorganisms.

The probiotic according to the invention is administered by adding to the corresponding feed for cattle and small cattle, pigs, poultry, in an amount of 500 to 1000 g / 1000 kg.

Efficacy and tolerability tests have shown that the product is well absorbed by animals and contributes to the restoration and regulation of the balance of the microbial intestinal ecosystem. The content of live bacteria in the finished feed is lxl0 ⁹ cfu / kg. All bacterial strains involved here are isolated from environmentally friendly natural sources and are not genetically modified.

Claims (3)

A livestock probiotic comprising a microbial complex and a plant substrate, characterized in that it comprises a microbial complex consisting of Lactobacillus acidophilus ^ XMCC 8242, Lactobacillus helveticus ^ WMCC 8269, Lactobacillus delbrueckii subsp. bulgaricus NBIMCC 8244, Lactobacillus delbrueckii subsp. lactis NBIMCC 8250, Streptococcus thermophilus NBIMCC 8253, Enterococcus faecium NBIMCC 8270 in an amount of 1x10 ⁹ colonies / g and vegetable substrate obtained from sunflower meal, containing crude protein from 25 to 30.0% by weight, carbohydrates from 8.0 - 11.0 %, fat from 0.3 to 0.6%, fiber from 8.0 to 15.0%, and free amino acids 39.0 to 40.0%, such as lysine, methionine, cystine, threonine, leucine, methylcystine, asparagine acid, serine, glutamic acid, glycine, alanine, valine, isoleucine, phenylalanine, histone, arginine. A method for producing a probiotic for animal husbandry according to claim 1, comprising fermenting a substrate of plant origin with a microbial complex, characterized in that the sunflower flour is dissolved in water to a dry matter content of 8-12%, and at temperatures of 45-52 ° C and pH 6-9 is enzymatically treated with a proteolytic enzyme selected from the group of serine proteases in an amount of 0.05 to 0.15% by volume of the substrate and a cellulotic enzyme selected from cellulase, phytase in an amount of 0.05 to 0.15%, after cooling the mixture to 37-40 ° C, fermentation with a starter culture of Lactobacillus acidophilus NBIMCC 8242, Lactobacillus helveticus ^ BXMCC 8269, Lactobacillus delbrueckii subsp. bulgaricus NBIMCC 8244, Lactobacillus delbrueckii subsp. lactis NBIMCC 8250, Streptococcus thermophilus NBIMCC 8253, Enterococcus faecium NBIMC 8270 imported in an amount of 0.05 to 0.1% by volume, and carrying out a fermentation process for 6-20 hours, during which neutralization of organic acids is applied , accumulating during the process, then, the enriched product is cooled and subjected to spray drying at a temperature drop from 185 - 190 ° C to 90 C to a dry matter content of 94 - 97%. Use of a probiotic according to claim 1, as a food additive for animal feed.