Utilize este identificador para referenciar este registo: http://hdl.handle.net/10400.22/3371
Título: Optimization of a microplate-based assay to assess esterase activity in the alga Pseudokirchneriella subcapitata
Autor: Machado, Manuela D.
Soares, Eduardo V.
Palavras-chave: Bioassay
Fluorescein diacetate (FDA)
Growth phase
Microplate assay
Metabolic activity
Selenastrum capricornutum
Data: 2013
Editora: Springer
Relatório da Série N.º: Water, Air, & Soil Pollution; Vol. 224, Issue 1
Resumo: The present work describes the optimization of a short-term assay, based on the inhibition of the esterase activity of the alga Pseudokirchneriella subcapitata, in a microplate format. The optimization of the staining procedure showed that the incubation of the algal cells with 20 μmolL−1 fluorescein diacetate (FDA) for 40 min allowed discrimination between metabolic active and inactive cells. The shortterm assay was tested using Cu as toxicant. For this purpose, algal cells, in the exponential or stationary phase of growth, were exposed to the heavy metal in growing conditions. After 3 or 6 h, cells were subsequently stained with FDA, using the optimized procedure. For Cu, the 3- and 6-h EC50 values, based on the inhibition of the esterase activity of algal cells in the exponential phase of growth, were 209 and 130 μg L−1, respectively. P. subcapitata cells, in the stationary phase of growth, displayed higher effective concentration values than those observed in the exponential phase. The 3- and 6-h EC50 values for Cu, for cells in the stationary phase, were 443 and 268 μgL−1, respectively. This short-term microplate assay showed to be a rapid endpoint for testing toxicity using the alga P. subcapitata. The small volume required, the simplicity of the assay (no washing steps), and the automatic reading of the fluorescence make the assay particularly well suited for the evaluation of the toxicity of a high number of environmental samples.
Peer review: yes
URI: http://hdl.handle.net/10400.22/3371
ISSN: 0049-6979
Versão do Editor: http://link.springer.com/article/10.1007/s11270-012-1358-3
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