Characterisation of the fluorescence from freshwater, planktonic bacteria.

Amino acid-like fluorescence has been used as an indicator of biological activity in wastewater effluent and in natural waters, and can be detected using fluorescence spectroscopy. Little or no work has been able to state conclusively that these so called 'amino acid-like' fluorophores are associated with proteins present as a result of bacterial activity. This work aims to ascertain whether bacteria are one possible cause of these 'amino acid-like' peaks observed in natural waters. In addition, fluorescence derived solely from one bacterial source was determined as a function of the growth time and temperature. The bacterium Pseudomonas aeruginosa was isolated from the urban River Tame, Birmingham, UK, and planktonic bacteria were grown in sterile, sealed glass jars, in 100 ml growth media. Bacteria were grown at 11, 25 and 37 degrees C, over a maximum of 10 days. A 3D Excitation-Emission Matrix (EEM) plot was generated from fluorescence analysis of the samples. Both tryptophan and tyrosine-like fluorescence, resembling that observed in natural and waste waters, was observed in these samples, indicating that observed fluorescence signals from aquatic systems in the literature were of biotic origin. Significant differences in fluorescence signals were obtained from planktonic cells grown at different temperatures. At 25 and 37 degrees C, cells were found to produce predominantly tryptophan-like fluorescence, with some tyrosine-like fluorescence also detected. A further unknown fluorophore was also detected (emission wavelength of approximately 460 nm, with three excitation centres at 225, 260 and 390 nm), likely to be a bacterially produced metabolite. At 11 degrees C, a more environmentally realistic temperature in temperate environments, quantitative and qualitative differences were observed in fluorescence signals when compared with the higher temperatures, indicating that laboratory observations conducted at higher temperatures may not be easily used to interpret environmental processes.