The cold growth phenotypes and transcriptional activation of cold stress adaptation genes was evaluated amongst L. monocytogenes strains from human listeriosis cases, food products and associated production environments. Significant cold growth phenotypic variation was observed during growth of such strains in rich (BHI) as well as chemically defined minimal (MDM) nutrient conditions. While all twenty analyzed strains grew in BHI at 4°C, only eight of these strains, mostly those recovered from human listeriosis cases, were also able to grow in MDM under similar cold stress. The cold growth phenotypes observed in BHI were used to define two categories of five strains each, which either displayed enhanced and poor cold tolerance relative to the rest of the strain collection. The first group (GP1) consisted of strains characterized by short lag times, whilst the second group (GP2) comprised of strains displaying prolonged lag times before growth resumption during incubation in BHI cultures at 4°C. Transcription level activation of sigB, cspA and pgpH gene expression associated with cold stress exposure in a selection of GP1 and GP2 strains was assessed. Despite similar cold dependent sigB transcript induction between these two strain groups, there were significant observed in cold stress dependent induction of cspA and pgpH transcripts. Cold tolerant GP1 strains displayed relatively higher transcriptional activation of cspA and pgpH after cold stress exposure compared to the cold sensitive GP2 strains. This study highlights strain variability in cold stress tolerance phenotypes, as well as in strain capacity to activate specific cold adaptation gene expression responses. In addition the study also shows that enhanced and poor cold growth phenotypes are associated with particular strain capacity to activate important cold stress gene expression responses upon transition of L. monocytogenes into low temperature environments.