SU570640A1 - Method of storing pure cultures of methanoxidizing bacteria - Google Patents

Description

one

The invention relates to the field of microbiology and may be used to store pure cultures of methane-acidifying bacteria in museum collections.

In microbiological practice, various methods of storing pure cultures of microorganisms are used.

A known method of storing pure cultures of methane-acidifying bacteria by nuteum of regular transfers (once every two weeks) of these microorganisms on solid or liquid nutrient media in an atmosphere of methane with oxygen (air).

The disadvantage of this method is that the storage of methane-oxidizing bacteria in the active state and by their frequent subcultures is associated with the contamination of the cultures during storage by extraneous microflora, mainly fungal microorganisms, which penetrate into the cultures through moist cotton plugs of test tubes or vessels. methane-air mixture. In addition, during storage of microorganisms by the method of reseeding, a significant change in their initial properties often occurs.

The aim of the invention is to preserve the purity of the cultures and their physiological activity.

This is achieved by keeping pure cultures of methane-oxidizing bacteria in a mineral liquid nutrient medium in the atmosphere of a mixture of methane and oxygen or methane.

and air that is supplied under pressure. At the same time, the partial pressure of methane is 2-4 atm, and the oxygen pressure is 5-7 atm, or the partial pressure of methane is 2-4 atm, and the air pressure is 25

35 atm

Example 1. Suspension of a pure culture of methane-oxidizing bacteria Methylosinus trichosporium, pc. 20 grown in liquid

NMS mineral medium with shaking at a rocking chair at 30 ° C, is poured into test tubes of 5 ml of suspension (concentration - 10 cells / ml) each, subject to the conditions of sterility. The tubes are closed with cotton plugs.

and placed in a metal vessel that can withstand gas pressure up to 200 atm. The vessel is hermetically sealed, methane is fed into it from cylinders under a pressure of 3 atm, and then air under a pressure of 35 atm. Gas pressure

monitored with a pressure gauge. The reservoir is then disconnected from air and methane sources and stored at room temperature. After the storage period, the gas from the vessel is slowly (for 20

30 min) let out, slightly opening the valve, closing one of the outlet channels of the vessel.

Example 2. The process is carried out as in Example 1, but instead of air, pure oxygen is injected into the vessel to a partial pressure of 7 atm.

Culture analysis of Methylosinus trichosporium pcs. 20 after 9 months of storage shows that during storage under pressure of methane with air, the concentration of bacteria in the suspension does not change, contamination of cultures with extraneous microflora is not observed, the cells of the studied bacteria begin to grow a day after sowing them in a 1MS liquid medium and give a good .

Claims (3)

1. A method of storing pure cultures of methane-acidifying bacteria in a mineral liquid In a nutrient medium in an atmosphere of methane and oxygen or methane and air, this is because in order to preserve the purity of the cultures and their physiological activity, the gas mixture is fed under pressure. 2. The method according to claim 1, wherein the partial pressure of methane is 2-4 atm, and the oxygen pressure is 5-7 atm. 3. The method according to claim 1, wherein the partial pressure of methane is 2-4 atm, and air pressure 25-35 atm.