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The physical and nutritional requirements of the antibiotic-producing slime mold Physarum gyrosum were examined to develop a liquid medium for this myxomycete. Liquid culture is desired to expedite a useful scale of production of antibiotic materials for ease of isolation and structure study. Culture conditions were selected to favor antibiotic production rather than maximum growth. The medium devised consisted of 0.010 M potassium phosphate buffer (pH 6.0), 2% bakers' yeast, and 0.2% glucose and was supplemented with either 10(-7) M hemoglobin (preferred) or 2.0 ml of live Escherichia coli per 100 ml of culture medium grown to a steady-state population in nutrient broth. The slime mold, which contained some E. coli carried along with the inoculum, was allowed to grow as a surface plasmodium at 20 degrees C in the dark with weekly subculturing for stocks or for 10 days for antibiotic production. P. gyrosum produced the same antibiotic materials when grown in liquid medium as it did when grown on agar plates. A seeded plate disk assay against Bacillus cereus was employed to follow antibiotic activity.

Growth of Physarum gyrosum on Agar Plates and in Liquid Culture