RU2252956C2 - Probiotic additive and method for production thereof - Google Patents

Abstract

FIELD: biotechnology and agriculture, in particular probiotic-containing compound feed for domestic animals, birds and fish.

SUBSTANCE: claimed additive is prepared by mixing of subtilis B-2250 and/or Bacillus licheniformis B-2252 biomass and carrier-sorbent and moisture capacitive filler as auxiliary substances. As carrier-sorbent hydrophilic and hydrophobic substances are used, and as moisture capacitive filler cation exchanging resins are used in predetermined ratio of dry mass. Probiotic additive in dry microcapsulated form is obtained by capillary sorptive drying of component mixture up to finished product humidity of 8-25 %.

EFFECT: probiotic additive with stable properties capable to improve feed conversion, to increase weight increment and to reduce animal mortality.

2 cl, 2 tbl, 4 ex

Description

The invention relates to biotechnology in the feed industry, specifically to obtain a probiotic supplement intended to increase the digestibility of feed as a result of improved digestion and the prevention of diseases of the gastrointestinal tract (GIT) of various etiologies, as well as to improve the adaptability of the use of the additive in the production and use of animal feed.

In modern food biotechnology, there is a serious problem of reducing the productivity of food production due to the death of animals, poultry, fish, especially young animals, from various gastrointestinal diseases.

To a large extent, the occurrence of these diseases is caused by a violation of the ecobiomicrococenosis of the gastrointestinal tract due to a drop in the level of bifidomicroflora of the intestines of the host organism and, accordingly, in the intensity of its immunity, including mucosal immunity.

The latter is a consequence of the activation of pathogenic and conditionally pathogenic microflora that enter the gastrointestinal tract with poor quality food, as a result of the infectious process, unsatisfactory epizootic conditions, excessive antibiotic and chemotherapy, the effect of endo and exotoxins, etc. The use of a probiotic feed additive helps to normalize ecobiomicrocenosis and reduce economic losses by maintaining the livestock population and increasing live weight.

Known liquid probiotic additive "Bicado" (TU-92.2900-017-02067968-95) containing lactulose and used in the cultivation of piglets by adding it to animal feed.

Known methods for producing liquid bifidogenic additives (RF patent dated 30.07.99, No. 2165712, publ. 04/27/01) based on bifidobacteria, lactulose enriched with non-protein nitrogen in the form of calcium hydroxide and urea, as well as additives "Bifidofan" on basis of lignin, lactulose and bifidobacteria.

All these probiotic supplements are not very effective in severe infections, intoxications, contamination of feed, and low-tech use.

A known method of obtaining a dry tablet form of a probiotic supplement containing a recombinant strain of Bacillus subtilis (Romashova NG and other Biotechnology 2001, No. 2, p. 64-69), lactose with a protective acid-resistant coating of tablets from AFC applied in a "pseudo-boiling layer "

The disadvantage of this method is that the use of the obtained probiotic additives requires special permission due to the presence of a genetically modified strain in it, in addition, it is not technologically advanced in the industrial production of animal feed.

The noted drawbacks determine the relevance and feasibility of developing a probiotic supplement that can increase feed digestibility, improve digestion processes, increase the immunity of animals, poultry and fish, improve the adaptability of the use in animal feed production.

The closest to the proposed invention should be considered a known method for producing a probiotic preparation based on Bacillus subtilis, according to which a tablet form or microcapsules is obtained using activated carbon, carbolong, lactulose, acetylftamelcellulose or aluminum phosphate gel as a sorbent. As auxiliary substances, calcium steorate, talc and / or starch, and / or aerosil, glucose, milk powder and / or methyl cellulose. The finished form of this drug is intended for the treatment and prevention of gastrointestinal diseases in humans (RU 2182009 C1, class A 23 K 1/165, 04/27/02).

When developing a probiotic additive, the tolerance of its constituent components was taken into account, with particular attention paid to the creation of such a preparative form of the additive that is able to ensure the stability of properties at all stages of its preparation, storage and use, as well as manufacturability and ease of use.

The technical result achieved by the implementation of the invention is to increase the efficiency of food production by increasing the viability and survival of young (juvenile) animals, poultry, fish, and increasing their average weight.

The task of developing a probiotic supplement capable of providing the desired result was solved by creating a dry microencapsulated form of a probiotic supplement containing spore-forming bacteria Bacillus subtilis B-2250 and / or Bacillus licheniformis B-2252, hydrophilic and hydrophobic aerosils of grades A and AM (TU 6-18- 185-84) and ion-exchange cation exchangers of the KB-4P-2 and KU-2-8 grades (GOST 20298-78) in the following percentages by weight of dry matter:

- biomass of Bacillus subtilis B-2250 - (0.5-6.0)%;

- and / or Bacillus licheniformis B-2252 - (0.5-6.0)%;

- Aerosil hydrophilic grade A - (0.5-2.5)%;

- Aerosil hydrophobic grade AM - (0.5-2.5)%;

- resin-cation exchange resin ion brand KB-4P-2 - (9-40)%;

- resin-cation exchange resin ion grade KU-2-8 hrs - (50-85)%;

In the proposed probiotic additive, as a result of a high-intensity dispersing mechanical action and the occurring capillary-sorption processes of interaction of the surfaces of microparticles of hydrophilic and hydrophobic aerosils, spore-forming bacteria and particles of resin-cationites, spore-forming bacteria Bacillus subtilis and / or Bacillus licheniformis are encapsulated in microcapsule a system that also includes particles of a moisture-absorbing sorbent (cation resins KB-4P-2 and KU-2-8 hs) and a guide ofobnogo Aerosil providing rheological properties of the system and an optimum flow of the heat-mass-exchange processes.

The spore-forming bacteria Bacillus subtilis B-2250 and / or Bacillus licheniformis B-2252, which are the active biologically active principle of the pro-biotic supplement, have the properties of a probiotic, have a pronounced antagonism to a wide range of pathogenic and conditionally pathogenic microorganisms, and are highly resistant to digestive juices gastrointestinal intestinal tract, and when an animal, bird or fish enters the gastrointestinal tract with food, colonize it and displace pathogenic and conditionally pathogenic bacteria, thereby restoring normal intestinal ecobiomicrocenosis ka.

The strain Bacillus subtilis B-2250 is protected by RF patent No. 2184774 with a priority of 09/21/01, for strain Bacillus licheniformis B-2252 filed application No. 2001126103 with a priority of 09/26/01.

The ion-exchange cation resins KB-4P-2 and KU-2-8 hs are a moisture-consuming filler and have a high sorption capacity. Available in accordance with GOST 20298-78. As part of the destruction-epitoxial system, microcapsules are shielded from adverse environmental interactions. At ratios from 1: 1 to 1: 5, toxins, salts of heavy metals, anabolics, xenobiotics, their metabolic products are sorbed on the surface of the intestine and, increasing intestinal motility, naturally removes them from the body.

Aerosil hydrophilic grade A is a structural substance of microcapsules.

Aerosil hydrophobic brand AM due to hydrophilic-hydrophobic interactions with microcapsules and particles of ion-exchange resins provides stability of the destruction-epitoxial system.

Aerosil hydrophilic grade A and aerosil hydrophobic grade AM are produced according to TU 6-18-185-84.

The value of (8-25)% of the residual moisture in the probiotic additive is determined by the GOST regulated amount of residual moisture in the feed into which the additive is introduced, and can vary in the indicated range.

Probiotic supplement is harmless, side effects and complications when it is used in accordance with the Manual, approved in the prescribed manner, is not observed. The products of animals, poultry, fish treated with the supplement are allowed to be used for food purposes without any restrictions.

Information confirming the possibility of carrying out the invention.

Example 1. The composition of the probiotic supplement with Bacillus subtilis B-2250

Component The content by weight of dry matter,
wt.% Liquid culture of Bacillus subtilis B-2250 1,0 Resin KB-4P-2 80.0 Resin KU-2-8chs 15.0 Aerosil brand A 2.0 Aerosil brand AM 2.0

Example 2. The composition of the probiotic supplements with Bacillus subtilis B-2250 and Bacillus licheniformis B-2252

Component The content by weight of dry matter, wt.% Liquid culture of Bacillus subtilis B-2250 0.5 Liquid culture of Bacillus licheniformis B-2252 0.5 Resin KB-4P-2 15.0 Resin KU-2-8chs 80.0 Aerosil brand A 2.0 Aerosil brand AM 2.0

Example 3. The composition of the probiotic supplement with Bacillus licheniformis B-2252

Component The content by weight of dry matter, wt.% Liquid culture of Bacillus licheniformis B-2252 1,0 Resin KB-4P-2 15.0 Resin KU-2-8chs 80.0 Aerosil brand A 2.0 Aerosil brand AM 2.0

Example 4. The implementation of the method of producing probiotic supplements.

Obtaining a probiotic additive is carried out in a highly intensified mixer-grinder with a predominantly convective bias mode with a fill factor of 0.4-0.5. The pre-weighed portions of a moisture-absorbing sorbent and a finely divided filler are loaded into the mixer tank and mixed for at least 2.5 minutes. When the mixer is operating, the liquid culture of Bacillus subtilis B-2250 and / or Bacillus licheniformis B-2252 with a flow rate of OD-0.2 dm ³ · min ^{-1 is} transferred from the measuring device by gravity or by a peristaltic pump. Upon completion of the transfer of liquid culture, mixing is continued for at least 3 minutes for further dispersion and microencapsulation of the liquid culture and its simultaneous dehydration by capillary-chemical method. At the end of the mixing process, the probiotic additive is discharged into the collection tank with the mixer working.

The finished probiotic additive is a mechanical mixture of a microencapsulated liquid culture of Bacillus subtilis B-2250 and / or Bacillus licheniformis B-2252 with a moisture-absorbing sorbent with a value of moisture and biological activity, regulated by the ratio of liquid culture / moisture-absorbing sorbent, with an inhomogeneity coefficient of not more than 8% and stable physical and mechanical characteristics.

In accordance with the proposed method, 10 series were prepared, the characteristics of which confirm the high reproducibility of the method for producing probiotic additives and the stability of its quality indicators.

The effectiveness of the proposed probiotic supplements based on Bacillus subtilis B-2250 and Bacillus licheniformis B-2252 was tested on newborn calves and pigs with acute intestinal diseases against the background of a difficult epizootic situation on the farm. Animals of the control groups were kept according to the zootechnical technology accepted at the farm. The calves and piglets of the experimental groups were additionally given the proposed probiotic supplement orally with a small amount of water 30 minutes before feeding in a single dose per head for 25 g of calves and 30 g of piglets three times a day for three days. Observations showed that in the experimental groups, one day after giving the probiotic supplement, the general condition of the animals improved, and after another two days all the animals were practically healthy. The death of animals in the experimental groups was not. In the control groups, the condition of the animals remained severe, mortality was 12% in calves and 28% in piglets.

The proposed probiotic supplement based on Bacillus subtilis B-2250 was tested in the Biserovsky fish factory in the Moscow region on juvenile trout. Conducted 6 courses of probiotic supplements from May to October 2002. The results are shown in table 1.

Table 1. Test results of probiotic supplements on juvenile trout.

date May, 23rd 21st of June July, 12 August 14 August, 26th September 18th October 15th Experience, g 0.3 4.69 13.0 20.54 28.6 52.78 74.13 Control g 0.3 4.75 11.5 16,0 24.56 42.18 60.21

As a result of the use of probiotic additives in experimental cages 1193 kg of trout were grown. In control cages, where the additive was not used, 969 kg were grown. The increase in live weight of trout in the experiment compared with the control amounted to 224 kg (23% more). The income, with the price of planting material 250 rubles / kg and the cost of a probiotic supplement 246 rubles, amounted to 56,000 rubles.

The proposed probiotic supplement, please call “Subtilis dry microencapsulated”.

Claims (2)

1. A probiotic additive containing the biomass of spore-forming bacteria Bacillus subtilis, a sorbent carrier and an auxiliary substance, characterized in that it contains a biomass of bacteria Bacillus subtilis B-2250 and / or Bacillus licheniformis B-2252, aerosil hydrophilic brand as a sorbent carrier A and hydrophobic brand AM, excipients - cation exchange resin-ion exchange grade KB-4P-2 and KU-2-8 hrs, with the following ratio of components by weight of dry matter, wt.%:  Bacillus subtilis B-2250 biomass 0.5-6.0  and / or biomass of Bacillus licheniformis B-2252 0.5-6.0  Aerosil hydrophilic grade A 0.5-2.5  Aerosil hydrophobic grade AM 0.5-2.5  ion-exchange resin-cation exchange grade KB-4P-2 9-40  resin-cation exchange resin ion grade KU-2-8 hrs 50-85. 2. A method of producing a probiotic additive, comprising mixing the biomass of the spore-forming bacteria Bacillus subtilis, a sorbent carrier and auxiliary substances and dewatering the resulting mixture, characterized in that the bacterial biomass Bacillus subtilis B-2250 and / or Bacillus licheniformis B- is used 2252, the adsorbent carrier is hydrophilic aerosil grade A and hydrophobic brand AM, excipients are ion exchange cation resins of the KB-4P-2 and KU-2-8 hs grades, the mixture is dehydrated by capillary-sorption drying moisture content in the final product 8-25%.