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Figure S1 - Impact of Sublethal Levels of Environmental Pollutants Found in Sewage Sludge on a Novel Caenorhabditis elegans Model Biosensor

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posted on 2012-10-03, 02:28 authored by Debbie McLaggan, Maria R. Amezaga, Eleni Petra, Andrew Frost, Elizabeth I. Duff, Stewart M. Rhind, Paul A. Fowler, L. Anne Glover, Cristina Lagido

Worm numbers per well in replicate independent experiments with the PE328 strain. Average number of worms per 96-well plate column (n = 8 wells; equivalent to one experimental technical replicate) taken from 2 independent PE328 experiments at the 4 h exposure timepoint (Figure 1E). After the luminescence readings, nematodes were pipetted out of the well (75 µl) and placed on a microscopic slide for counting. An extra 75 µl of S complete plus 0.01% tween was then added to the well and the first tip/pipette used to remove any remaining nematodes from the well for counting. Prior to discarding the tip, it was checked under the microscope for any worms adhering to it (0.01% tween minimises this) and counts were added together to obtain the total count for the well. In this example columns 1, 3, 5 were exposed to 1% DMSO control (average ± SEM: 30.0±1.8, experiment 1; 29±0.8, experiment 2) and columns 2, 4 and 6 to 0.5% SSE (30.2±1.3, experiment 1; 28.2±1.1, experiment 2). The minimum and maximum values observed in a well were: 18, 45 (experiment 1) and 17, 45 (experiment 2). No significant differences in nematode numbers were found between the 6 columns in each experiment (comprising 8 wells each): P>0.05 in experiment 1 and 2. Reducing the number of replicate wells within each column to 5 for the purpose of statistical testing had no effect and differences in worm numbers remained nonsignificant in both experiments (P>0.05). One way ANOVA applied to squared root transformed data.

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