Rapid toxicity testing based on yeast respiratory activity

Haubenstricker, M. E.; Meier, Peter G.; Mancy, K. H.; Brabec, Michael J.

doi:10.1007/bf01701786

Cited by 16 publications

(7 citation statements)

References 13 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Mitochondria have provided data that have been correlated with cytotoxicity parameters derived from cell cultures and whole organisms (Blondin et al, 1987;Knobelock et al, 1990). Microorganisms, like yeasts (Haubenstricker et al, 1990) and bacteria (Sikkena et al, 1992;Carneiro de Melo et al, 1996), have also been used in toxicity tests that resulted in toxicity correlations concerning energy metabolism and cell viability (Sikkena et al, 1992;Donato et al, 1997b). A strain of the thermophilic eubacterium Bacillus stearothermophilus has been used as a model to study the cytotoxicity of several lipophilic drugs (Luxo et al, 2000;Rosa et al, 2000;Monteiro et al, 2003) and pollutants (Donato et al, 1997a,b;Martins et al, 2003) and a remarkable parallelism of effects on this prokaryotic model and other model systems (Donato et al, 1997a) has been observed.…”

Section: Introductionmentioning

confidence: 99%

Toxicity of methoprene as assessed by the use of a model microorganism

Monteiro

Jurado

Moreno

et al. 2005

Toxicology in Vitro

View full text Add to dashboard Cite

Methoprene is an insect juvenile growth hormone mimic, commonly used as a pesticide. Although widely used for the control of several pests, toxic effects on organisms of different phyla have been reported. These events triggered studies to clarify the mechanisms of toxicity of this insecticide putatively involved in ecological issues. Here we show the effect of methoprene on the normal cell growth and viability of a strain of the thermophilic eubacterium Bacillus stearothermophilus, previously used as a model for toxicological evaluation of other environment pollutants. Respiration studies were also carried out attempting to identify a putative target for the cytotoxic action of methoprene. Cell growth was affected and a decrease of the number of viable cells was observed as a result of the addition of methoprene to the growth medium, an effect reverted by the presence of Ca(2+). Methoprene also inhibited the redox flow of B. stearothermophilus protoplasts before the cytochrome oxidase segment, an effect further studied by individually assessing the enzymatic activities of the respiratory complexes. This study suggests that methoprene membrane interaction and perturbation of cell bioenergetics may underlie the mechanism of toxicity of this compound in non-target organisms.

show abstract

Section: Introductionmentioning

confidence: 99%

Toxicity of methoprene as assessed by the use of a model microorganism

Monteiro

Jurado

Moreno

et al. 2005

Toxicology in Vitro

View full text Add to dashboard Cite

show abstract

“…Saccharomyces cerevisiae has been used previously to assess toxicity through the use of an amperometric gas diffusion (oxygen) electrode, which quantifies changes in culture respiration (1,7,8,9,17). Alternatively, the effect of a compound on S. cerevisiae cultures was measured directly through inhibition of maximum growth rates (2,10).…”

mentioning

confidence: 99%

Design and Application of a Biosensor for Monitoring Toxicity of Compounds to Eukaryotes

Hollis

Killham

Glover

2000

Appl Environ Microbiol

View full text Add to dashboard Cite

Here we describe an alternative approach to currently used cytotoxicity analyses through applying eukaryotic microbial biosensors. The yeast Saccharomyces cerevisiae was genetically modified to express firefly luciferase, generating a bioluminescent yeast strain. The presence of any toxic chemical that interfered with the cells' metabolism resulted in a quantitative decrease in bioluminescence. In this study, it was demonstrated that the luminescent yeast strain senses chemicals known to be toxic to eukaryotes in samples assessed as nontoxic by prokaryotic biosensors. As the cell wall and adaptive mechanisms of S. cerevisiae cells enhance stability and protect from extremes of pH, solvent exposure, and osmotic shock, these inherent properties were exploited to generate a biosensor that should detect a wide range of both organic and inorganic toxins under extreme conditions.Many luminescent bacterial biosensors have been produced which detect a wide range of pollutants while simultaneously assessing bioavailability in environmental samples. Saccharomyces cerevisiae has been used previously to assess toxicity through the use of an amperometric gas diffusion (oxygen) electrode, which quantifies changes in culture respiration (1,7,8,9,17). Alternatively, the effect of a compound on S. cerevisiae cultures was measured directly through inhibition of maximum growth rates (2, 10). However, such toxicity assays are time-consuming and expensive in comparison to luminometry analysis. Walmsley et al. (18) created an S. cerevisiae biosensor that induces green fluorescent protein expression on exposure to genotoxic agents. The luminescent biosensor designed in the present study works on a different principal (reduction in reporter gene product activity) and complements the biosensor designed by Walmsley et al. (18) by detecting a wide range of toxins, not just genotoxic agents.For the novel S. cerevisiae biosensor described here, a luminescence detection system was constructed using firefly luciferase (luc) from Photinus pyralis. The firefly luciferase light reaction relies on ATP being supplied by actively metabolizing cells. This dependence on endogenous energy supplies enables a luciferase assay system to report directly on cell health upon exposure to toxins. The luciferin substrate for P. pyralis luciferase is an amphipathic molecule with a charged carboxyl group at physiological pH. This prevents easy passage of luciferin across cell membranes, leading to problems during the exogenous addition for in vivo assays. This was overcome through the development of a novel assay system where the biosensor preparation was acidified after exposure to the toxicant and before luminescence quantification. MATERIALS AND METHODSStrains, media, and chemicals. S. cerevisiae strain W303-1B 15 trp1-1 can1-100 ade2-1 ura3-1) (16) was the host for the chromosomal luciferase-expressing construct. Chromosomal insertion was carried out using a published method (5). S. cerevisiae was grown in synthetic complete medium (15)

show abstract

“…Respect to the methodology already established [1,2,5,9,10] other variations have been introduced: 1) Using the serial dilution plate technique, cells counting was performed on Petri dishes with culture media of different composition ( Table 1). The composition allowing obtaining a number of cells suitable for respirometric test was selected.…”

Section: Improvements In Sensitivity: Lysis Of the Cellular Wallmentioning

confidence: 99%

“…Since 1990s biosensors have been studied and developed to test and screen inorganic and organic substances [1][2][3]. A biosensor consists of 1) a biological recognition element, namely the biomediator, and 2) a transduction element, namely the sensor.…”

Section: Introductionmentioning

confidence: 99%

Sensor with Intact or Modified Yeast Cells as Rapid Device for Toxicological Test of Chemicals

Dragone¹,

Frazzoli²,

Grasso³

et al. 2014

JACEN

View full text Add to dashboard Cite

Aerobic catabolism of S. cerevisiae (cell respiration) is a rapid, cost-effective, and reproducible toxicological endpoint of the whole cells biosensor. To increase the signal intensity, a protocol for the immobilization and modification of the yeast cells is described. In particular, the enzymatic treatment of the immobilized yeast cells allows removing the cell wall and obtaining structurally modified cells namely spheroplasts. Both immobilization and exposure of sensitive cells like spheroplasts confirmed to improve the method's sensitivity vs. the chemicals. The present paper reports the test of different chemicals (including Mercury and wood preservative like Tanalith) present in consumer products, performed both by sensor with intact and modified whole cells.

show abstract

Rapid toxicity testing based on yeast respiratory activity

Cited by 16 publications

References 13 publications

Toxicity of methoprene as assessed by the use of a model microorganism

Toxicity of methoprene as assessed by the use of a model microorganism

Design and Application of a Biosensor for Monitoring Toxicity of Compounds to Eukaryotes

Sensor with Intact or Modified Yeast Cells as Rapid Device for Toxicological Test of Chemicals

Contact Info

Product

Resources

About