RU2249615C2 - Strain clostridium cellobioparum useful in probiotic preparation for ruminants - Google Patents

Abstract

FIELD: biotechnology, in particular probiotics for stock-raising.

SUBSTANCE: Strain Clostridium cellobioparum is isolated from caw punch dominant microflora and identified using conventional microbiological methods. Strain Clostridium cellobioparum ferments complex feedstuff polysaccharides, activates processes of digestion, volatile fat acid synthesis, whereby increases cow dairy productivity. Preparation obtained using strain of present invention contains 10⁹-10¹⁰/g of viable cells.

EFFECT: feedstuff additive of improved characteristics.

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Description

The invention relates to the microbiological industry, biotechnology, the production of probiotics and can be used in animal husbandry.

Literary sources reported the significance and isolation of ruminants from the rumen into a pure culture of bacteria of the genus Clostridium, which decompose complex polysaccharides. However, information on their use in the production of probiotics is not provided [1].

A known strain of Lactobacillus acidophilus var. coccoideus M-86, a fermenting starch used in the manufacture of the ruminolact probiotic for ruminants [2].

The aim of the invention is to obtain a new strain of Cl. cellobioparum KEM-537, with the ability to decompose cellulose, starch, which make up the bulk of ruminant feed.

The strain isolated from the dominant microflora of the rumen of cows and is characterized by the following features.

Morphological and cultural properties

Cells are in the form of sticks with a size of 0.8 × 7 μm, are sporadic, form spores located terminally, gram-positive, mobile. The cultivation of the strain is carried out on an agar medium of Omelyansky of the following composition, g / l: KNH ₄ HPO ₄ 0.2; KH ₂ PO ₄ 0.1; CaCl ₂ 0.03; peptone 0.1; MgSO ₄ 0.05; CaCO ₃ 0.5; filter paper 20.0; agar 20.0.

Crops are incubated at (34 ± 1) ° C for 24-36 hours under anaerobic conditions. On a nutrient medium colonies of beige color deep with a diameter of 1-2 mm are formed. In a liquid medium of the same composition form a uniform turbidity or a weak precipitate.

Physiological and biochemical properties of the strain

Anaerobe. Does not dilute gelatin, does not form lecithinase, urease, lipase, hydrogen sulfide. It ferments: cellulose, starch, cellobiose, glucose, mannitol, lactose, dulcite, fructose, galactose, sucrose, trehalose, xylose, ribose, maltose, mannose, melibiosis. The optimum growth temperature is 30-37 ° C, grows at 10 ° C, and can withstand boiling.

The pH value is 6.8-7.0. The strain is non-pathogenic, does not show toxic and toxigenic properties.

The strain is stored in a viscous nutrient medium with the addition of 0.15% agar under anaerobic conditions at a temperature of (4 ± 2) ° C.

Example 1

The strain is grown at a temperature of (34 ± 1) ° C for 16-18 hours on a medium of the following composition, g / l: KH ₂ PO ₄ 0.4; MgSO ₄ 0.04; NaCl 0.6; (NH ₄ ) ₂ SO ₄ 0.9; microcrystalline cellulose 1.4; whey up to 1 l; pH 6.8-7.0. The nutrient medium is sterilized at 1 atm (121 ± 1) ° C for 20 minutes. The prepared medium is cooled to (34 ± 1) ° C and inoculated with a daily culture of Clostridium cellobioparum KEM-537 1-5% by volume of medium.

After the fermentation process is completed, the resulting product is used to prepare a paste-like or dry preparation.

Example 2

The paste is prepared by centrifuging the culture medium. The bacterial mass is mixed with serum concentrate (application No. 2000125930 from 04/26/01) in a ratio of 1: 10-1: 12, getting the number of viable cells 10 ⁹ -10 ¹⁰ in 1 g.

Example 3

A dry preparation is obtained by mixing biomass with an adsorbent in a ratio of 1:10. Flour, starch, glucose, and whey powder can be used as adsorbent. The number of living microbial cells reaches 10 ⁹ -10 ¹⁰ in 1 g of the drug.

The drug is used as a biologically active feed additive for cows.

Strain Cl. cellobioparum KEM-537 has a high enzymatic activity of polysaccharide cleavage, enhances digestive processes, the assimilation of feed nutrients, the synthesis of volatile fatty acids, which are the main sources of energy for ruminants and are precursors of milk constituents.

Sources of information

1. Pivnyak I.G., Tarakanov B.V. Microbiology of the digestion of ruminants. - M .: Kolos, 1982, 247 p.

2. Patents of the Russian Federation No. 2154102 C2, 1993; No. 1541250 A1, 1990.

Claims (1)

The strain Clostridium cellobioparum KEM-537, used to obtain a probiotic for ruminants, is in the collection of the Museum of Pure Microorganism Cultures of the Vologda State Dairy Academy named after N.V. Vereshchagin.