A method for isolating and culturing anaerobic bacteria

Abstract

The invention discloses a separation culture method of anaerobic bacteria. The method comprises the following steps: preparing a culture medium; filling nitrogen before sterilizing the culture medium; carrying out enrichment culture of a strain; and carrying out separation culture on the strain, wherein the separation culture on the strain adopts a way of alternate inoculated culture by a liquid culture medium and a solid culture medium, and the liquid culture medium inoculates the solid culture medium by capillary glass tube puncture, a cylindrical anaerobic culture cavity with a smooth inoculating face is formed in the solid culture medium, and round holes formed on the surface of the solid culture medium are sealed after inoculation by adopting an unset culture medium so as to form a sealed anaerobic culture cavity, and the inner wall of the anaerobic culture cavity is a culture face of anaerobic bacteria. According to the method disclosed by the invention, the anaerobic culture cavity with the smooth inoculating face is formed in the solid culture medium to provide an anaerobic environment for culturing anaerobic bacteria, so that the use of anaerobic equipment is reduced, the separating process is simplified, the cost in the separating process is lowered, and the time of acquiring single strain is shortened.

Description

technical field [0001] The invention relates to the field of microorganisms, in particular to a method for isolating and cultivating anaerobic bacteria in oil production reuse water, oil field sewage and oily sludge. Background technique [0002] In the petrochemical industry, corrosion caused by microorganisms leads to huge economic losses. It is reported in the literature that more than 75% of the corrosion in oil wells, and 50% of the failures in buried pipelines and cables come from microbial corrosion (mainly the sulfate reduction process); The loss caused by corrosion to the oil field is as high as 200 million yuan. [0003] As the development of most onshore oilfields enters the middle and later stages, the amount of water injection, steam injection and oilfield reinjection water is increasing year by year in order to increase the recovery rate, and the water content of the produced crude oil is also increasing. In order to prevent microbial corrosion, it is necessar...

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Problems solved by technology

However, this invention utilizes the method of mixing the medium of unsolidified agar (temperature higher than 50°C) and the bacterial enrichment solution to inoculate. For anaerobic bacteria whose growth environment temperature is lower than 50°C, the inoculation process described in this invention will affect Activity of the microorganism to be isolated, even leading to its death