Development of a novel, FTIR (Fourier transform infrared spectroscopy) based, yeast bioassay for toxicity testing and stress response study.
Fourier transform infrared spectroscopy (FTIR) was used to analyze the metabolomic alterations caused to yeast cells by four chemical compounds: ethanol, sodium hypochlorite, sodium chloride and sulfur dioxide, each tested at five different concentrations. The complex of four stressing agents at different concentrations, inducing cell mortalities ranging from 1% to 100%, has given the opportunity to prove that FTIR can individuate the presence of a stress before the cells start dying. A series of "Stress Indexes" was calculated with an expressly designed "R" script, to estimate the level of stress induced by the chemical agents at different concentrations. These estimation procedures allowed the direct comparison of the stress induced by the four agents at different concentrations. The response spectra, calculated as difference between the spectrum of the cells under stress and that of the cells maintained in water, showed different shapes in the diverse experimental conditions, suggesting a specificity of the response and the possibility to classify it. The contribution of five different spectral regions (fatty acids, amides, mixed zone, carbohydrates and typing region) could be calculated separately, gaining additional information on the stressing effects. Spectral alterations were detected at concentrations as low as 10% ethanol, 20 ppm bleach, 1 M NaCl and 100 mg L(-1) SO(2). These preliminary findings suggest that FTIR technology and a series of simple algorithms can be employed to study response of cells to various stressing situations, not limited to chemical agents. The ease and rapidity of the FTIR analysis suggest that this approach could be used as a bioassay in several applications and particularly in ecotoxicology and in environmental microbiology.
Corte
Laura
L
Department of Applied Biology-Microbiology, University of Perugia, Borgo 20 Giugno, 74, I-06121 Perugia, Italy.
Rellini
Paolo
P
Roscini
Luca
L
Fatichenti
Fabrizio
F
Cardinali
Gianluigi
G
D000431:Ethanol / Q000633:toxicity; D055432:Metabolomics; D012441:Saccharomyces cerevisiae / Q000187:drug effects* / Q000378:metabolism; D012965:Sodium Chloride / Q000633:toxicity; D012973:Sodium Hypochlorite / Q000633:toxicity; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*; D013312:Stress, Physiological / Q000187:drug effects*; D013458:Sulfur Dioxide / Q000633:toxicity; D018675:Toxicity Tests
D013458:Sulfur Dioxide; D000431:Ethanol; D012965:Sodium Chloride; D012973:Sodium Hypochlorite