Detection of the viable but nonculturable state in Escherichia coli O157:H7

Enterohemorrhagic Escherichia coli O157:H7 (EHEC) is recognized as a frequent cause of gastroenteritis ranging from mild to severe bloody diarrhea. The Shiga-like toxin produced by EHEC can result in hemolytic uremic syndrome, now the major cause of acute kidney failure in children in the United States. Inadequately cooked beef is most commonly implicated in the transmission of EHEC, although only a small fraction of cattle appear to harbor the organism. In several studies EHEC positive herds were detected only in the summer months correlating with the occurrence of human infections. Numerous E. coli strains have been shown to enter a viable but nonculturable (VBNC) state as a result of environmental stresses, including low temperature. Using traditional plating methods and the BacLight Molecular Probe, we monitored EHEC strains incubated in river water (RW) and artificial sea water (ASW) at temperatures of 5C and 25C for their entry into the VBNC state. EHEC strains remained culturable for over 40 days in both ASW and RW incubated at 25C. In ASW, these levels were higher than a non-EHEC control. At 5C, the number of culturable EHEC cells dropped gradually in both RW and ASW. Using the BacLight Molecular Probe, we were able to demonstrate that these cells, though not culturable, were viable indicating entry into the VBNC state. Our study suggests that temperature and not salinity is the primary signal for entry into this state.