Use of bioluminescent Escherichia coli O157:H7 to study intra-protozoan survival of bacteria within Tetrahymena pyriformis

Cooper, Alison A.A.; Taylor, Elaine L.; Salisbury, Vyvyan C.; Nelson, Shona

Abstract

A method was developed that enabled real-time monitoring of the uptake and survival of bioluminescent Escherichia coli O157 within the freshwater ciliate Tetrahymena pyriformis. Constitutively bioluminescent E. coli O157 pLITE27 was cocultured with T. pyriformis in nutrient-deficient (Chalkley's) and in nutrient-rich (proteose peptone, yeast extract) media. Non-internalised bacteria were inactivated by addition of colistin, indicated by a decline in bioluminescence. Protozoa were subsequently lysed with Triton X-100 which lead to a further drop in bioluminescence, consistent with release of live internal bacteria from T. pyriformis into the colistin-containing environment. Bioluminescence measurements for non-lysed cultures indicated that internalised E. coli O157 pLITE27 cells were only slowly digested by T. pyriformis, in both media, over the time period studied. The results suggest that bioluminescent bacteria are useful tools in the study of bacterial intra-protozoan survival. © 2003 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

• Journal Article Type: Article
• Publication Date: Jun 6, 2003
• Journal: FEMS Microbiology Letters
• Print ISSN: 0378-1097
• Electronic ISSN: 1574-6968
• Publisher: Oxford University Press (OUP)
• Peer Reviewed: Peer Reviewed
• Volume: 223
• Issue: 1
• Pages: 95-99
• DOI: https://doi.org/10.1016/S0378-1097%2803%2900349-5
• Keywords: bioluminescence, protozoan, intracellular survival, escherichia coli O157:H7
• Public URL: https://uwe-repository.worktribe.com/output/1069818
• Publisher URL: http://dx.doi.org/10.1016/S0378-1097(03)00349-5
• Additional Information: Additional Information : This was the first publication to evidence bioluminescence from live bacteria within protozoan cells. The work reported here led to a NERC funded Environment and Human Health project and also to a BBSRC and DTI funded project using bioluminescent bacterial biosensors for rapid pre-screening of chemotherapy efficacy in mamellian cells.