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A Simplified Method of Obtaining Differential and Total Viable Anaerobic Counts of Rumen Bacteria Employing Sealed Small-Diameter Glass Tubing¹

Departments of Dairy Science and Microbiology, University of Kentucky, Lexington

ABSTRACT

Two major obstacles encountered when attempting to culture rumen bacteria are the difficulty in maintaining anaerobic conditions throughout, and the limited number of samples which can be handled within a reasonable length of time. Sections of small glass tubing (4 mm. outside diameter, cut into 12-in. lengths) are employed in much the same way as are Burri tubes, in an effort to overcome these obstacles. These hold 1 ml. of culture medium and allow about 2 in. of gas space at both ends of the column of medium.

The glass tubing sections are placed in a canister and sterilized in an autoclave before use. A 2-ml. sterile hypodermic syringe, fitted with a sterilized rubber adapter, is used to pull the medium into the tubing. The syringe is first filled with CO₂ gas and then fitted to one end of a section of glass tubing. The other end of the tubing is inserted in the mouth of a test tube containing medium with the desired dilution of whole rumen content, and the CO₂ in the syringe is exhausted through the tubing to remove all air.

¹ The investigation reported in this paper is in connection with a project of the Kentucky Agricultural Experiment Station and is published with the approval of the Director.