A modification of the method using cellulose overlay agar to isolate and purify cellulolytic cytophagas from enrichment culture

We report here on a modification of the cellulose overlay agar method for isolating and purifying cellulolytic cytophagas from enrichment cultures. We call it the "back-door" method, and it overcomes two existing problems. First, it prevents recontamination with organisms growing on the agar surface. Second, it permits purification of cellulolytic cytophagas that are unable to utilize glucose or that are accompanied by a rapidly growing contaminant with spreading habit. Pure cultures of cellulolytic cytophagas were obtained from five enrichment cultures of nine soil samples examined. Two limitations are that (i) the cytophaga must be able to penetrate the overlay and (ii) some noncellulolytic cytophagas may also penetrate the overlay to yield cocultures. In conclusion, the back-door method can be used both to isolate and purify cellulolytic cytophagas from soils, using only a cellulose-based medium. Key words: back-door method, cellulolytic cytophagas, cellulose overlay agar, cellulolysis.