Identification and Control of Bacterial Contamination in Callus Cultures of Ulmus americana

Bacillus macerans Schardinger appeared on culture media and forceps used in serial transfers of Ulmus americana callus tissue after several contamination-free transfers and may have originated as an endophyte in the leaf tissue used as an explant. Bacteria remained viable on forceps stored for several weeks in 95% ethanol whether or not the excess was burned off in a flame from an alcohol lamp. Bacteria were eliminated from forceps treated similarily with 85% ethanol. The bacterium on artificially contaminated forceps remained viable after immersion up to 4 hr in either 95%, 85%, 80%, or 70% ethanol with or without flaming. Artificial contamination was eliminated, either by autoclaving for 20 min at 121C (185.8F) or exposure to a bunsen burner flame for 6-8 sec. Bacillus macerans remained viable in both naturally and artificially contaminated ethanol at dilutions of 95%, 85%, 80%, and 70%. Thus, forceps may be contaminated by latent bacteria in callus or contaminated ethanol. Ethanol may become contaminated by storage of nonsterile forceps and airborne spores introduced during routine, septic procedures.