In situ and laboratory studies of bacterial survival using a microporous membrane sandwich

A new device and procedure for the study of bacterial survival in an aquatic environment are described. The device uses two appressed presterilized microporous membranes to expose a bacterial cell suspension to the environment at a cell concentration that closely resembles those levels found in natural aquatic ecosystems. The device has been used under laboratory controlled conditions and in situ to study and compare bacterial survival times. In laboratory studies, Escherichia coli and Streptococcus faecalis survived the longest at 12 degrees C, pH 5, and in the presence of iron or calcium ions and cysteine. Cells in mid-stationary growth phase survived longer than those in mid- or late-logarithmic phase, whereas those maintained for a year or more as stock cultures survived for shorter period of time than did recent environmental isolates. In situ studies indicate that 5% of the starting number of E. coli and S. faecalis cells may survive longer than 96 h at 16 degrees C in potable lake water, whereas survival times in polluted lake water were approximately 12 h.