A NOVEL TOXICITY FINGERPRINTING METHOD FOR POLLUTANT IDENTIFICATION WITH lux-MARKED BIOSENSORS

Abstract

A novel technique is described for the identification and quantification of environmental pollutants based on toxicity fingerprinting with a metabolic lux-marked bacterial biosensor. This method involved characterizing the toxicity-based responses of the biosensor to seven calibration pollutants as acute temporal-dose response fingerprints. An algorithm is described to allow comparisons of responses of an unknown pollutant to be made against the calibration data. This is based on predicting pollutant concentration at each of six different time points over the course of a 5-min assay. If the prediction is consistent between the unknown pollutant and a calibration pollutant at the 95% test level, this is considered to be a positive identification. All seven calibration pollutants could be successfully distinguished from each other with this technique. Environmental samples, individually spiked with single concentrations of pollutants, were compared in this way against the calibration pollutants. An 83% identification success was achieved, with no false positives at the 95% test level. This is a simple and rapid technique that potentially can be applied to monitoring of industrial wastewater or as a screening tool for regulators.

On line biomonitors used as a tool for toxicity reduction evaluation of in situ groundwater remediation techniques

“…Gu et al (1999) and Gu and Gil (2001) used differently reacting recombinant Escherichia coli strains in an on line system for identification of substances in a mixture. Turner et al (2001) also used the combination of the lux gene with an E. coli strain to use the fingerprint like reactions of the E. coli to the substances to detect them in a mixture as did Kobatake et al (1995) who wanted to use bacteria tests instead of chemical methods for the detection of benzene and its derivatives in environmental samples. Disadvantage for on line measurement in this work was the premise of an incubation of the sample with the bacteria at 37 • C for several hours.…”

The use of multiple-strain algal sensor chips for the detection and identification of volatile organic compounds

“…By application of non-selective biological components combined with the analysis of response patterns, possibly, the number of detectable targets can be increased without concomitantly increasing the number of probes (biological components) accordingly. This principle was shown by Turner et al (2001), who detected several toxicants with a luminescent bacterial biosensor by analysis of luminescence fingerprints. Since the identification of toxic compounds by analysing the response patterns of different algal strains was demonstrated here with only three strains and two VOCs, an increase in the number of chip-immobilized strains and VOCs to be tested seems appropriate.…”

Contamination and screening level toxicity of sediments from remediated and unremediated wetlands near Sydney, Australia

“…The toxicity of pore‐water samples was assessed by the luminescent bacterium E. coli HB101 pUCD607 lux biosensor assay [6,28]. The luminescent bacterium E. coli HB101 is marked with the lux CDABE genes, isolated from Vibrio fischeri , using the multicopy plasmid pUCD607.…”

“…Luminescent bacterial biosensors, including MicrotoxH (Strategic Diagnostics) and other lux-marked biosensors, are rapid and convenient tools for determining the toxicity of contaminants in water, soil, and sediment samples [3][4]. Metabolic lux biosensors have been applied successfully to a range of organic and inorganic contaminants and shown to be sensitive, reproducible, and not limited by a narrow pH range [5][6][7][8][9][10][11]. As it is difficult to assess toxicity of whole sediment to various species in terms of bioavailability, pore water has often been used in toxicity screening since it is a key exposure route for contaminants to benthic organisms [12][13][14].…”