Investigation of the effect of temperature on cytotoxicity in poikilothermic cells, exploiting biosensor technology

Abstract

The effect of temperature on the sensitivity of poikilothennic cells to toxicant exposure was investigated, with particular attention to the relationship between temperature induced changes in cellular activity, and cell sensitivity to toxicants. Temperature was shown to have a significant influence on the metabolic activity and sensitivity to toxicants ofthree types ofpoikilothenns: E. coli, a consortium of cells isolated from activated sludge (ASBC), and a genetically modified bioluminescent fish cell line (BF-2Ilucl). The influence of temperature on the ASBC and BF-21/ucl cell sensitivity to toxicants appeared to be related to its effects on toxicant uptake and reactivity. However temperature induced changes in E. coli metabolic activity were shown to have a pronounced influence on its sensitivity to toxicants. The increased metabolic rate supported by higher temperatures was associated with decreases in E. coli sensitivity to narcotic toxicants as were the increases in E. coli metabolic activity that resulted from changes in respiratory substrate solution composition. Subsequent biosensor and growth assays demonstrated that E. coli responded to low concentrations of phenolic toxicants by increasing it respiration rate at the expense of growth. This suggests that the protective effect ofincreased metabolic activity at higher temperatures was related to the energetic costs of toxicant exposure. Additionally an impedance spectroscopy assay was developed and showed that 3,5-DCP caused only limited disruption ofE. coli membrane integrity. This study demonstrated for the first time that temperature effects on E. coli metabolic activity and on E. coli sensitivity to toxicants are directly linked. Further work is needed to develop a fuller understanding ofhow E. coli metabolic activity influences its sensitivity to toxicants.