Use of a methylene blue azide medium for isolation of enterococci

A methylene blue azide medium (MBA), developed by Schaedler, Dubos, and Costello to isolate enterococci from the gastrointestinal tract of animals, was evaluated. This was done by comparing the isolation of enterococci from feces and saliva on the medium. Fifty-two catalase-negative, gram-positive cocci from human feces isolated from MBA were classified as enterococci. All strains grew in S F, 6.5% NaCl, and streptomycin broths, and all fermented mannitol. The isolates were provisionally subdivided into Streptococcus faecalis and S. faecium groups. S. faecalis-like strains fermented glycerol and pyruvate aerobically and produced acid in Snyder's medium (initial pH, 4.8). The S. faecium group fermented raffinose. Among all strains, several tests were variable. These included growth at 45 C, in 0.1% tellurite and in methylene blue milk. Three methods were employed to isolate and identify enterococci from the oral cavity. Direct streaking of MBA with saliva failed to produce any growth on the medium. Two other methods, with the use of various selective broths to promote the recovery of oral enterococci, failed to produce any bacteria capable of growing on MBA. The MBA-isolated fecal strains and oral viridans streptococci were generally indistinguishable on Mitis-Salivarius and K F agars. In experiments with fecal material, no gram-negative bacilli were found among the isolates selected. The MBA medium was judged as a high selectivity-low yield medium, and may provide a means of separating fecal and nonfecal enterococci.     

Studies on Survival of Bacteria: Rhythmic Response of Microorganisms to Freeze-Drying Additives

Various materials were mixed with suspensions of Serratia marcescens and other organisms. Samples were removed and frozen at intervals after mixing; the number of cells that survived both freeze-drying and exposure to air varied rhythmically as a function of time between mixing and freezing. When assayed before or immediately after drying there were essentially no fluctuations. The response was evident only when these dried samples were exposed to air. In a typical experiment, the number of cells surviving in the sample frozen 30 sec after adding propyl gallate was at least 10 times that in samples frozen either 20 sec earlier or 20 sec later. Other "peaks" in survival were observed at approximately 125 and 450 sec, but the times at which the peaks were observed were not consistent from one experiment to the next. Although we have been unable to control or predict the time at which maxima in resistance occur, we have shown that the phenomenon does occur with Escherichia coli and Saccharomyces cerevisiae as well as with S. marcescens. Furthermore, a rhythmic response also was obtained after a change in pH or cell concentration. It appears that microorganisms respond physiologically and synchronously to changes in their environment, and some of these responses have survival value.