Exogenously provided carnitine (beta-hydroxy-L-tau-N-trimethyl aminobutyrate) was found to stimulate aerobic growth of enterohaemorrhagic Escherichia coli O157:H7 in a medium of inhibitory osmotic strength. Its osmoprotective ability is comparable with that of betaine. As carnitine is an important compound in mammalian tissues, it is suggested that it might play a role in the growth of the pathogen on low water activity (aw) meat products. Using specific uptake mutants of E. coli K-12, it was established that, under osmotic stress, carnitine accumulates in the cytoplasm following import through the ProP and ProU transport systems. Betaine and carnitine also protect E. coli cells while growing anaerobically at inhibitory osmolarity. Under these conditions, an E. coli K-12 strain with lesions in both proP and proU accumulates low levels of L-carnitine but fails to accumulate betaine when these compounds are supplied in the external medium. This is probably a result of uptake of L-carnitine by the secondary transporter CaiT. The caiT gene forms part of the caiTABCDE operon which encodes the carnitine pathway, and is transcribed during anaerobic growth in the presence of carnitine. However, further experiments revealed that the carnitine pathway, including CaiT, does not play a significant role in osmoregulation of E. coli during anaerobiosis. Together, the results indicate that ProP and ProU are the sole transport systems involved in carnitine influx, both in aerobically and anaerobically osmotically stressed E. coli cells.