Strain of microorganism lactobacillus buchneri 66 for ensilage of maize green mass and preserving flattened grain

FIELD: biotechnology, microbiology, agriculture.

SUBSTANCE: the strain Lactobacillus buchneri 600 is isolated from barley grains at milkwax stage of ripeness. The strain Lactobacillus buchneri 600 is registered in 05. 08. 2002 year in the All-Russian State collection of microorganism strains used in veterinary science and animal husbandry at number Lactobacillus buchneri VGNKI 02.08.04-DEP and deposited in the collection 000 "Biotrof". Invention provides the more effective multiplication of the strain in maize ensilaging green mass and preserving flattened grains with enhanced formation of lactic acid, inhibition of putrid microflora that allows preparing fodder from vegetable raw with enhanced quality. Invention can be used in fodder production for ensilage maize green mass and preserving flattened grains.

EFFECT: valuable properties of strain.

3 tbl, 2 ex

 

The invention relates to forage production, in particular the production of a drug for silage corn and canning rolled grain, and relates to a new strain of bacteria for this drug.

A known strain of Lactobacillus plantarum 60 for silage feed described in the patent of Russian Federation №2168909 (for example, from 14.04.2000), IPC C 12 N 1/20 (12 N 1/20, 12 R 1/25, And 23 To 3/00), registered in the State register of inventions of the Russian Federation on 20 June 2001.

Lactobacillus plantarum 60 deposited in the collection of the all-Russian research Institute for agricultural Microbiology (ARRIAM) 06.04.2000, under N 78 D.

The specified strain used to prepare the drug for silage feed. When this bacteria strain effectively multiply in ferment green mass, synthesize lactic acid and inhibit putrefactive microflora

However, the drug is prepared on the basis of Lactobacillus plantarum 60, lacks enzymatic activity for high-quality silage corn and canning grain rolled oats, as it contains little protein and soluble sugars.

The specified strain selected as a prototype.

The task of the claimed invention to provide a new strain of bacteria that is able to more effectively multiply ferment in the green mass of corn, as well as conserve the SQL flattened grain, to synthesize lactic acid, to suppress putrefactive microflora and get better food from vegetable raw materials.

This problem is solved due to the fact that for silage corn and canning rolled grain used bacterial strain Lactobacillus buchneri 600.

Lactobacillus buchneri 600 isolated from barley grain milky-wax ripeness.

Lactobacillus buchneri 600 registered 05.08.2002, all-Russian state collection of strains of microorganisms, used in veterinary and animal husbandry number of Lactobacillus buchneri VGNKI 02.08.04-DEPT and stored in the collection LLC Biotrop”.

Lactobacillus buchneri 600 is characterized by the following morphological-cultural, physiological and biochemical characteristics.

Cells have the form of short rods, single or in chains. Gram-positive, catalytically. Colonies on nutrient glucose minor, point, deep. Not diluting gelatin, casein is not split. Have a low level of amylase activity. The strain has an antagonistic activity against the fungi Penicillium sp., Aspergillus niger and A. flavus, the yeast genera Torula, Pichia and Saccharomyces. Heterofermentative. Form a gas from glucose and gluconate. The optimum growth - 25-37°C. Grow at 15°With, do not grow at 45°C. Grow in the presence of 6% NaCl. Fermented sugars: arabinose, fructose, glucose, maltose, ribose, xylose, SL is Bo fermented the raffinose. Not fermented to cellobiose, rhamnose, sorbitol, mannitol, galactose, maltose, sucrose, lactose.

Optional gone anaerobic. On meat-peptone agar grows poorly, forming a subtle drop increase over the bar. On meat-peptone agar - growth weak. On wort agar with chalk forms a characteristic colonies (with area enlightenment chalk) - lenticular with smooth edges or rounded surface, pasty consistency, white or cream shade. The colony diameter 0,6-1,2 see Grow in the presence of 6% NaCl.

Milk containing methylene blue (0.1%), no growth. Culture can withstand a temperature of 60°C for 30 minutes, but growth is weak. On media containing 40% bile, the strain grows well. Strain needs growth stimulants.

Energy cyclotourisme and the main products of fermentation of lactic acid bacteria determined in the process of their cultivation in wort with subsequent determination of organic acids by the method of Wigner.

Table 1 presents the results of a study of the growth and synthesis of organic acids by strains of bacteria (L. plantarum 60 - prototype, L. buchneri 600 - patented strain).

Table 1

The breeding activity of lactic acid bacteria and the formation of acids
Options experienceThe number is about bacteria million/mlThe acid content, % lactic/acetic
Age, days
1213510
L.plantarum 6060010003,7/0,53,3/0,46,5/0,33,4/0,5
L.buchneri 6006509503,6/0,43,4/0,56,6/0,33,6/0,5

As can be seen from the table. 1, the strain L.buchneri 600 has about the same speed reproduction, as the strain L.plantarum 60, and the same capacity to produce mainly lactic acid.

Table 2 presents the results of studying the antagonism of the above strains of lactic acid bacteria against putrefactive bacteria found in the silo.

Table 2

Antagonistic properties of lactic acid bacteria (zone of inhibition in mm)

OptionsThe culture test
Century megatheriumCentury mesentericusB. subtilisCentury mycoides
Control0000
L.plantarum 6022,022,020,025,0
L.buchneri 60021,023,019,026,0

As can be seen from the table. 2, the strain L.buchneri 600 has about the same antagonistic properties against putrefactive bacteria of various kinds, like strain L.plantarum 60, and the same ability to form zones of inhibition.

Strain L.buchneri 600 is supported on the wort with chalk and re-seeding after 3 months. Stored in the refrigerator at a temperature of +(4...5)°C.

Example of getting a drug for silage using Lactobacillus buchneri 600.

Preparation of uterine culture. In a sterile nutrient medium containing (in%vol.) molasses - 1.0, Biotin - 0,5, meat and bone meal to 0.9, sodium chloride - 6,0, water water - the rest, making the bacteria in the amount of 1-2% vol. and grow within 24-30 hours at a temperature of 25-30°C.

Industrial cultivation of the strain in the fermenter. Prepare production environment, sterilized, cooled to 30-32°and contribute 4% vol. seed (mother culture)obtained by the above method. The cultivation is carried out at 28-30°C for 20-24 hours at a pressure of 0.3 to 0.5 atmospheres without blowing air. The stirrer incorporate immediately after application of inoculum for 3-5 min, and ZAT is m every 3-5 hours of cultivation and before sampling.

Ferment for silage is a liquid light brown with sediment nutrient medium and contains lactic acid bacteria in the amount of 100 million/ml. Activity of lactic acid bacteria (acidity by Turner) - 25-30°C.

An example of using the obtained bacterial ferments.

For silage use corn in the stage of milky-wax ripeness. Chopped vegetable mass in the amount of 1.5 kg, put in a glass vessel with a volume of 3 l, add bacterial yeast in an amount of 0.01% by weight of green mass. The mass ferment thoroughly stirred, RAM, banks cover with plastic wrap and pour the melted wax.

Similarly, use sourdough for canning rolled grain collected in the stage of wax ripeness. Silage quality determined after 90 days.

The results of using the proposed strain compared with its prototype is presented in table. 3.

Table 3

The indicators of the composition of the feed
IndicatorsCorn with the addition of:Rolled grain with the addition of:
No additivesL.

plantarum 60
L.

buchneri 600
Without

doba is OK
L.

plantarum 60
L.

buchneri 600
PH5,204,604,55the 5.654,854,50
Lactic acid, %3,253,804,203,303,50the 3.65
Acetic acid, %0,751,000,900,450,440,48
Butyric acid, %000000
The amount of red. sugars0,741,301,650,750,750,90
Ammonia, mg/kg136,094,080,0120,0105,090,0
Crude protein, %to 12.013,214,113,214,014,8

The bacterial strain Lactobacillus buchneri 600 VGNKI 02.08.04-DEPT for ensilage of green mass of corn and canning rolled grain.



 

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