RU1784641C - Strain of bacterium lactobacillus plantarum having nitrareductase activity and used for siloing fodders - Google Patents

Abstract

Usage: agriculture, feed production. SUMMARY OF THE INVENTION: Lactobaclllus plantarum VKPM B-5337 strain of lactic acid bacteria isolated from silage. It has the ability to reduce nitrates, a wide range of fermentable carbohydrates, antagonistic activity against causative agents of silage damage. The introduction of the strain into the silage mass improves the quality of the resulting feed. 2 tab.

Description

The invention relates to the microbiological industry, in particular to the preparation of a bacterial preparation for silage of feeds and relates to a new strain of lactic acid bacteria having nitrate reductase activity.

In the process of spontaneous fermentation of plant mass, partial or complete decomposition of nitrates occurs, which is largely due to the activity of nitrate-reducing microorganisms, the development of which reduces the quality of the resulting feed. The introduction of chemical preservatives that help to suppress unwanted microorganisms and preserve food nutrients often inhibits the decomposition of nitrates in the silage.

The purpose of the invention is to provide a new strain capable of reducing nitrates and improving the quality of silage feed.

A new strain of Lactobacillus plantarum was isolated from silage prepared from cane ovule by deep plating of silage chatter on Petri dishes with agarized cabbage medium to which 10% ethanol and 2% CaCO3 were added.

The resulting strain of Lactobacillus plantarum 3766 deposited in the All-Union collection of industrial microorganisms under the number VKPM B-5337.

The strain is characterized by the following properties.

Morphological and cultural properties.

Short (0.8-1 x 2.8-3.2 microns) gram positive sticks, single or pairwise linked, rarely - chains of 3-4 cells; motionless, do not form a dispute. When deep plated on MRS-arap with CaCO3 form a clear zone around the colonies; in a liquid medium (stationary culture in MRS) turbidity is first observed

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medium, tiny growth along the tube wall, then a homogeneous white precipitate forms. Injection growth in MRS-arape is uniform, does not reach the surface of the medium.

Phaeological and biochemical properties.

Aero-tolerant microorganism, acid-resistant, fermentation type metabolism. It grows at 15 ° C, at 45 ° C there is no growth. The optimum growth temperature is 2b-30 ° C, pH 6.5-6.8. -h -,.,

Step 1 doesn’t have activity, gelatin does not dilute, does not contain cytochrome-containing respiratory systems, does not form ammonia from arginine, reduces nitrates to nitrites on modified BBL medium, does not form gas from glucose and Na gluconate.

When fermented, glucose forms predominantly lactic acid, as well as acetic acid. Ethanol does not form.

In addition to glucose, he uses sucrose, arabinose, cellobiose, and esculin as a carbon source. Na-gluconate, lactose, maltose, mannitol, melecitose, melibiosis, raffinose, sorbitol, trehalose with accumulation of acids, weakly hydrolyzes starch.

For normal strain growth, substrates with complex organic forms of nitrogen (protein hydrolysates, amino acid mixtures), as well as vitamins and growth factors, are required.

The strain shows antagonistic activity against Pseudomonas aeruglnosa, Bacillus subtllls, Escherlchla coli, PenlcU Hum purpurogenum, Asperglllus terreus.

The strain is maintained in an active state by periodic (1 time in 20-30 days) transfers to a liquid wort, semi-agarized MRS medium with a glucose content reduced to 0.5%. CaCO3 (2%) was added to the storage media.

To extend the shelf life, the culture was lyophilized and stored at 4 ° C.

Strain L plantarum 3766 is neither toxic nor pathogenic ..

Example. The seed of the strain was grown in MRS liquid medium with an initial pH of 6.5 at 28 ± 2 ° C. Growth time 24 hours. Seed in an amount of 5% by volume was added to the medium. Containing corn extract (1.5%), molasses (2%), pH 6.5. The culture was grown in 2 L volumetric flasks at 28 ± 2 ° C for 48 hours. The crushed vegetable was treated with the obtained microbial suspension (at the rate of 0.01%)

mass, tightly packed in 3-liter containers, closed and stored at a temperature of 19 ± 2 ° C. At the same time, control options were put - without preservatives, with the chemical preservative VIC.

At certain time intervals, an analysis of the resulting silage was carried out using conventional methods.

The results of the analysis of silage samples are presented in table. 1,2. indicate that the introduction of the L. plantarum 3766 culture into the silage mass of corn and galega allows optimizing the ratio of organic acids, increasing the safety of feed nutrients, and reducing the nitrate content. The nitrite nitrogen formed as a result of nitrate reduction under conditions of high active acidity in the silo is unstable, quickly utilized and is not found in the finished silo.

Strain L plantarum 3766 has been proposed for use in feed silage to reduce nitrate nitrogen and improve silage quality.

The invention provides a number of advantages over other preservatives: a strain of microorganisms is obtained that is capable of reducing nitrates under certain conditions; the strain has antagonistic activity against the causative agents of silage damage, high acid formation energy, a wide range of fermentable carbohydrates, including pentose Sugars, starch, which leads to a positive effect of the L plantarum 3766 culture on microbiological and biochemical processes during the siloing of difficultly growing plants; the use of the proposed strain for silage of plant mass containing an increased amount of nitrate nitrogen allows one to lower the nitrate content below the MPC level while maintaining the high quality of the obtained feed.

The socially beneficial advantages of the proposed strain are that its use, unlike chemical preservatives, does not lead to environmental pollution and does not adversely affect the health of people and agricultural animals.

Claims (1)

SUMMARY OF THE INVENTION Bacterial strain Lactobaclllus plantarum VKPM B-5337, having nitrate reductase activity and used in feed silage. 51784641b Table 1 Characterization of corn silage after six months of storage In the initial green mass of corn, the nitrate content was 691.3 mg / kg. Characteristics of silage from galega after three months of storage table 2