Continuous pollution monitoring using Photobacterium phosphoreum

Photobacterium phosphoreum is a marine bacterium which is used extensively as a bioluminescent indicator of pollutants, where the presence of toxicants diminishes light output. To evaluate the utility of cell immobilisation in continuous toxicity testing, the sensitivity of P. phosphoreum to five gelling agents was evaluated relative to the retention of bioluminescence in 3% NaCl-glycerol suspensions. Following storage at 4°C, the control cultures retained light output for up to 2 weeks before significant decline; alginate-glycerol suspensions were stable for up to 4 weeks and bioluminescence was detectable for up to 6 weeks. Cells stored in agar were no more stable than the control, whereas cells gelled in agarose and low-melting point agarose showed a significant decline in bioluminescence within 2 weeks of storage. Bioluminescence was totally retained in alginate-glycerol suspensions stored at -80°C for up to 12 weeks. P. phosphoreum was successfully immobilised in strontium alginate and showed a dose-related response to four of the five heavy metal ions, SDS and pentachlorophenol tested when responses were followed over a time-course. A flow-through system for Sr-alginate immobilised cells was developed and conditions for operation were optimised. When cells were exposed to a pulse of 4-nitrophenol or salicylate then the nutrient feed continued, bioluminescence declined in response (pulse of 4-6 min) to these pollutants then recovered to a new stable rate of decline which was faster than the pre-exposure rate. These results demonstrate the potential of using immobilised P. phosphoreum in a continuous flow-through system for real-time environmental monitoring of water quality.