Biomedical subjects

Gareth Lloyd-Jones

Publications and source records attributed to Gareth Lloyd-Jones.

5 recordsLinked to original sources

PubMed2006

Bacterial oxygenases: in vivo enzyme biosensors for organic pollutants.

An in vivo enzyme-based biosensor platform was developed that uses specific oxygenase enzymes to detect aromatic compounds in water. Bacteria capable of degrading highly reduced hydrocarbons initiate substrate oxidation using well-characterised oxygenase enzymes, which due to their specificity, stability and high activity can be applied in vivo as biosensor components. Oxygenase enzyme activity was determined in vivo using BD Oxygen Biosensor plates to measure oxygenase-mediated oxygen depletion in the presence of specific aromatic analytes. The BTEX family of compounds (benzene, toluene, ethylbenzene and the three isomers of xylene) were used as model compounds. Detection limits and sensitivity achieved using this approach (microM detection range) was similar to levels achieved with oxygen electrode-based and some recombinant DNA-based approaches. No significant signals were detected with hydrocarbons that were not substrates of the initial oxygenases.

Bacteria, Aerobic↗

PubMed2006

The Te-Assay: a black and white method for environmental sample pre-screening exploiting tellurite reduction.

We present the tellurite bioassay (Te-Assay) as an alternative approach for quantification of cell viability. The Te-Assay was developed to pre-screen environmental samples for potential bacterial toxicants in which the reduction of tellurite to tellurium is used as a metabolic marker; black phenotype development only occurs in metabolically competent bacteria capable of reducing tellurite (TeO(3)(2-)) to elemental tellurium. The black and white phenotypes equate to nonsignificant or significant impediment of normal metabolic processes, thus permitting the rapid visual assessment of the relative toxicity of environmental samples. Bacterial inocula were exposed in 96-well plates to arrays of diluted analytes or environmental samples before addition of a tellurite to assess cell health/viability. Toxicity was quantified as the analyte concentration at which a 50% reduction in blackness occurred (IC(50)) compared to control wells containing no added analyte. No proprietary strains or reagents are required for Te-Assay, in which characterised strains or recent environmental isolates performed equally well. Strain selection was independent of tellurite-resistance provided that tellurite was reduced intracellularly by active non-growing cells.

Bacteria↗

PubMed2006

Use of protoplasts from paired heterogenic bacterial species to detect tin contaminants: prospects for biosensor development.

Two different bacteria gave different respiratory responses to the test analytes, tributyl tin (TBT) and cadmium as expressed by positive sigmoid responses by Halomonas sp. (slope, +1.71 [TBT]; +1.76 [Cd]) and negative sigmoid responses by Bacillus pumilis (slope, -1.06 [TBT]; -0.59 [Cd]). The EC50 values determined from Hill plots for the response of Halomonas sp. to the TBT and Cd were 1 and 8.5 mM, respectively, which were lower by a factor of 10 than the corresponding values for B. pumilis. With protoplasts of B. pumilis there was a major shift in the signal from sigmoid negative to positive with TBT (+1.35) but not Cd (-0.5), while the signals with the remaining protoplast-analyte combinations remained unchanged. For all four protoplast-analyte combinations the EC50 values were in the order of 10-100-fold lower than those for their whole cell counterparts. When other analytes were tested the protoplasts gave a similar response to tin as for TBT, but detected copper and 2,4-dichlorophenol with similar signal profiles to Cd and with lower sensitivity. The difference in signal and higher sensitivity of the two species protoplast system towards TBT/tin compared to the other analytes tested, suggests that it may feasible to develop this approach for the detection of tin residues.

Bacillus↗

PubMed2006

A resazurin-based biosensor for organic pollutants.

A new rapid biosensor method employing the dye resazurin as an indicator of bacterial respiration has been developed to provide a rapid, facile and specific biosensor for environmental contaminants that does not rely on genetic modification techniques, is suitable for a high-throughput multiwell format, and is ideally suited to resource-constrained environmental monitoring situations. This whole-cell biosensor has been applied to the test analyte toluene using natural toluene-degrading bacteria as the biological component and is competitive with more complex recombinant approaches. The redox-driven biosensor is dependent on the catabolism of a specific compound, concomitantly reducing the redox indicator resazurin to provide the analytical signal in a whole-cell biosensor assay.

Biological Assay↗

PubMed2005

Quantification of the Pseudomonas population in New Zealand soils by fluorogenic PCR assay and culturing techniques.

The genus Pseudomonas contains fast-growing nutritionally versatile bacteria that are able to utilize a wide variety of carbon sources. The ubiquity of the genus has been highlighted by conventional microbiology and the genus is well represented in collections of cultured bacteria. Here we evaluate the Pseudomonas population in New Zealand soils by comparing a culture-independent (real-time PCR combined with fluorescent TaqMan technology) with a culture-dependent (Gould's S1) population estimate. We show that cultivated fluorescent pseudomonads are not numerically dominant and represent a small proportion of <1% of the total Pseudomonas population, and that the total Pseudomonas population itself represents only a small proportion of <1% of the total bacterial population.

Bacteriological Techniques↗