Revival and subsequent isolation of heat-injured bacteria by a membrane filter technique

Abstract

Cultures containing mixed flora from raw milk were heated at 62.8 C for 15, 20, 25, and 30 min. Dilutions were filtered through membrane filters, and the filters were incubated on Trypticase soy broth (TSB) and on TSB plus NaCl (TSBS). The TSB count indicated the total population which survived heating and included injured and uninjured cells. The colonies on TSBS indicated the uninjured cells and were marked by perforating the membrane near the colony. This membrane was then transferred to fresh TSB and incubated further. The injured organisms recovered and formed colonies which could be distinguished from previous colonies of uninjured organisms. Transfer counts on TSB were not substantially different from the initial TSB counts at 15, 20, 25, and 30 min of heating.