Recent Developments in Microbiotesting and Early Millennium Prospects

Blaise, C.; Gagné, François; Bombardier, Manon

doi:10.1007/978-94-011-4369-1_2

Cited by 9 publications

(9 citation statements)

References 37 publications

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“…The intensity of green fluorescence exhibited by live cells depends primarily on the esterase activity of the cell. Using flow cytometry, the reduction of esterase activity on the algal cells, after exposure to different toxicants, was evaluated (Arsenault et al 1993;Blaise and Menard 1998;Cote et al 1998;Blaise et al 2000;Franklin et al 2001;Regel et al 2002;Liu et al 2008;Debenest et al 2011). In the present work, a low-cost bioassay, performed in a 96-well microplate format, based on the inhibition of the esterases in P. subcapitata, was optimized.…”

Section: Discussionmentioning

confidence: 99%

“…As the toxic effect increases, the enzyme activity is reduced in a dose-dependent manner. Therefore, 1 h (Franklin et al 2001;Regel et al 2002), 3 h (Franklin et al 2005), 4 h (Blaise et al 2000), 24 h Regel et al 2002;Franklin et al 2005;Prado et al 2009;Debenest et al 2011), 27 h (Arsenault et al 1993), or 28 h assay (Gala and Giesy 1994), based on the evaluation of the toxic effect on the esterase activity of algal cells, measured by flow cytometry, was reported.…”

Section: Introductionmentioning

confidence: 99%

“…Esterases have been recognized as a potential useful biomarker and alternative endpoint for the detection of early responses in a variety of microorganisms (Dorsey et al 1989;Blaise et al 2000;Bitton 2005; Peperzak and Brussaard 2011) to different toxicants. Esterases were used to evaluate the toxicity of heavy metals, such as Cd, Cu, Hg, Pb, and Zn (Arsenault et al 1993;Snell et al 1996;Franklin et al 2001), organic compounds such as phenol and naphthol (Snell et al 1996), brominated flame retardant (Debenest et al 2011), pesticides and insecticides (Gilbert et al 1992;Snell et al 1996), and in natural samples like surface seawater (Gilbert et al 1992), freshwater sediments , interstitial water (Cote et al 1998), and acid mine drainage (Regel et al 2002).…”

Section: Introductionmentioning

confidence: 99%

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Optimization of a Microplate-Based Assay to Assess Esterase Activity in the Alga Pseudokirchneriella subcapitata

Machado

Soares

2012

Water Air Soil Pollut

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The present work describes the optimiza-tion 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 diac-etate (FDA) for 40 min allowed discrimination be-tween metabolic active and inactive cells. The short-term 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 subse-quently stained with FDA, using the optimized proce-dure. For Cu, the 3-and 6-h EC 50 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 concentra-tion 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 toxic-ity of a high number of environmental samples.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Optimization of a Microplate-Based Assay to Assess Esterase Activity in the Alga Pseudokirchneriella subcapitata

Machado

Soares

2012

Water Air Soil Pollut

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“…Since that time, the development of microbiotests has proceeded apace, and their importance is becoming ever greater in the determination of environmental pollution levels and risk assessment. Hitherto, organisms from all trophic levels have been applied to such purposes (Blaise et al 2000). Originally, these tests were applicable to only to the aquatic environment; now, there are microbiotests available for assessing the toxicity of soils and sediments (Casado-Martinez et al 2007).…”

Section: Introductionmentioning

confidence: 99%

Determination of EC 50 toxicity data of selected heavy metals toward Heterocypris incongruens and their comparison to “direct-contact” and microbiotests

Kudłak

Wolska

Namieśnik

2010

Environ Monit Assess

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The sensitivity of Heterocypris incongruens to selected heavy metal ions is discussed. Although the Ostracodtoxkit ® has been present on the market for a few years, data on its selectivity and sensitivity to toxicants is scarce; such data is indispensable when interpreting the results of sediment toxicity. The LC(50) and EC(50) results with the Ostracodtoxkit ® are compared with those obtained with other commercially available "direct-contact" tests (utilizing Hyalella azteca and Chironomus riparius) and microbiotests for assessing the toxicity of aqueous samples (Vibrio fischeri, Daphnia magna, and Selenastrum capricornatum). The sensitivity of H. incongruens to metal ions (Cd(2+)> >Hg(2+)> Cu(2+)>Cr(6+ )>Ni(2+) ≈ Mn(7+)>Zn(2+)>Pb(2+)>Li(1+ )> Fe(3+)) was found to be similar to that of H. azteca and of C. riparius. The Ostracodtoxkit ® has shown itself to be an efficient and reliable element of test batteries for toxicity determination.

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“…Enzymes, namely, esterases, have been recognised as potentially useful for the detection of early responses to toxicants in a variety of microorganisms (Dorsey et al 1989;Blaise et al 2000;Bitton 2005;Peperzak and Brussaard 2011). Esterase activity has been evaluated using fluorescein diacetate (FDA).…”

Section: Introductionmentioning

confidence: 99%

Use of a fluorescence-based approach to assess short-term responses of the alga Pseudokirchneriella subcapitata to metal stress

Machado

Soares

2014

J Appl Phycol

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This work explores the use of fluorescent probes to evaluate the responses of the green alga Pseudokirchneriella subcapitata to the action of three nominal concentrations of Cd(II), Cr(VI), Cu(II) and Zn(II) for a short time (6 h). The toxic effect of the metals on algal cells was monitored using the fluorochromes SYTOX Green (SG, membrane integrity), fluorescein diacetate (FDA, esterase activity) and rhodamine 123 (Rh123, mitochondrial membrane potential). The impact of metals on chlorophyll a (Chl a) autofluorescence was also evaluated. Esterase activity was the most sensitive parameter. At the concentrations studied, all metals induced the loss of esterase activity. SG could be used to effectively detect the loss of membrane integrity in algal cells exposed to 0.32 or 1.3 μmol L −1 Cu(II). Rh123 revealed a decrease in the mitochondrial membrane potential of algal cells exposed to 0.32 and 1.3 μmol L −1 Cu(II), indicating that mitochondrial activity was compromised. Chl a autofluorescence was also affected by the presence of Cr(VI) and Cu(II), suggesting perturbation of photosynthesis. In conclusion, the fluorescence-based approach was useful for detecting the disturbance of specific cellular characteristics. Fluorescent probes are a useful diagnostic tool for the assessment of the impact of toxicants on specific targets of P. subcapitata algal cells.

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Recent Developments in Microbiotesting and Early Millennium Prospects

Cited by 9 publications

References 37 publications

Optimization of a Microplate-Based Assay to Assess Esterase Activity in the Alga Pseudokirchneriella subcapitata

Optimization of a Microplate-Based Assay to Assess Esterase Activity in the Alga Pseudokirchneriella subcapitata

Determination of EC 50 toxicity data of selected heavy metals toward Heterocypris incongruens and their comparison to “direct-contact” and microbiotests

Use of a fluorescence-based approach to assess short-term responses of the alga Pseudokirchneriella subcapitata to metal stress

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