Development of an improved rapid enzyme inhibition bioassay with marine and freshwater microalgae using flow cytometry

Franklin, NM; Adams, MS; Stauber, JL; Lim, RP

Development of an improved rapid enzyme inhibition bioassay with marine and freshwater microalgae using flow cytometry

Franklin, NM Adams, MS Stauber, JL Lim, RP

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Publication Type:: Journal Article
Citation:: Archives of Environmental Contamination and Toxicology, 2001, 40 (4), pp. 469 - 480
Issue Date:: 2001-06-09

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Field	Value	Language
dc.contributor.author	Franklin, NM	en_US
dc.contributor.author	Adams, MS	en_US
dc.contributor.author	Stauber, JL	en_US
dc.contributor.author	Lim, RP https://orcid.org/0000-0002-4916-5688	en_US
dc.date.issued	2001-06-09	en_US
dc.identifier.citation	Archives of Environmental Contamination and Toxicology, 2001, 40 (4), pp. 469 - 480	en_US
dc.identifier.issn	0090-4341	en_US
dc.identifier.uri	http://hdl.handle.net/10453/3678
dc.description.abstract	A rapid toxicity test based on inhibition of esterase activity in marine and freshwater microalgae (Selenastrum capricornutum, Chlorella sp., Dunaliella tertiolecta, Phaeodactylum tricornutum, Tetraselmis sp., Entomoneis cf. punctulata, Nitzschia cf. paleacea) was developed using flow cytometry. Uptake of fluorescein diacetate (FDA) was optimized for each species by varying the substrate concentration, incubation time, and media pH. Propidium iodide (PI) was utilized to assess membrane integrity. The optimized FDA/PI staining procedure was then used to assess the toxicity of copper in short-term exposures (1-24 h). Esterase activity was a sensitive indicator of copper toxicity in S. capricornutum and E. cf. punctulata. As copper concentrations increased, esterase activity decreased in a concentration-dependent manner. The 3- and 24-h EC50 values (based on mean activity states) were 112 μg Cu L-1 (95% confidence limits 88-143) and 51 μg Cu L-1 (95% confidence limits 38-70) for S. capricornutum and 47 μg Cu L-1 (95% confidence limits 43-51) and 9.1 μg Cu L-1 (95% confidence limits 7.6-11) for E. cf. punctulata, respectively. This enzyme inhibition endpoint showed similar sensitivity to chronic growth rate inhibition in E. cf. punctulata (48-h and 72-h EC50 values of 17 and 18 μg L-1, respectively) but was less sensitive compared to growth for S. capricornutum (48-h and 72-h EC50 values of 4.9 and 7.5 μg L-1, respectively). For the other five species tested, inhibition of FDA fluorescence was relatively insensitive to copper, even at copper concentrations that severely inhibited cell division rate. These short-term bioassays that detect sublethal endpoints may provide a more rapid and cost-effective way of monitoring contaminant impacts in natural waters.	en_US
dc.relation.ispartof	Archives of Environmental Contamination and Toxicology	en_US
dc.relation.isbasedon	10.1007/s002440010199	en_US
dc.subject.classification	Toxicology	en_US
dc.subject.mesh	Copper	en_US
dc.subject.mesh	Fluoresceins	en_US
dc.subject.mesh	Esterases	en_US
dc.subject.mesh	Xenobiotics	en_US
dc.subject.mesh	Biological Assay	en_US
dc.subject.mesh	Flow Cytometry	en_US
dc.subject.mesh	Lethal Dose 50	en_US
dc.subject.mesh	Environmental Monitoring	en_US
dc.subject.mesh	Cost-Benefit Analysis	en_US
dc.subject.mesh	Eukaryota	en_US
dc.title	Development of an improved rapid enzyme inhibition bioassay with marine and freshwater microalgae using flow cytometry	en_US
dc.type	Journal Article
utslib.citation.volume	4	en_US
utslib.citation.volume	40	en_US
utslib.for	0602 Ecology	en_US
utslib.for	050102 Ecosystem Function	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
utslib.copyright.status	closed_access
pubs.issue	4	en_US
pubs.publication-status	Published	en_US
pubs.volume	40	en_US

Abstract:

A rapid toxicity test based on inhibition of esterase activity in marine and freshwater microalgae (Selenastrum capricornutum, Chlorella sp., Dunaliella tertiolecta, Phaeodactylum tricornutum, Tetraselmis sp., Entomoneis cf. punctulata, Nitzschia cf. paleacea) was developed using flow cytometry. Uptake of fluorescein diacetate (FDA) was optimized for each species by varying the substrate concentration, incubation time, and media pH. Propidium iodide (PI) was utilized to assess membrane integrity. The optimized FDA/PI staining procedure was then used to assess the toxicity of copper in short-term exposures (1-24 h). Esterase activity was a sensitive indicator of copper toxicity in S. capricornutum and E. cf. punctulata. As copper concentrations increased, esterase activity decreased in a concentration-dependent manner. The 3- and 24-h EC50 values (based on mean activity states) were 112 μg Cu L-1 (95% confidence limits 88-143) and 51 μg Cu L-1 (95% confidence limits 38-70) for S. capricornutum and 47 μg Cu L-1 (95% confidence limits 43-51) and 9.1 μg Cu L-1 (95% confidence limits 7.6-11) for E. cf. punctulata, respectively. This enzyme inhibition endpoint showed similar sensitivity to chronic growth rate inhibition in E. cf. punctulata (48-h and 72-h EC50 values of 17 and 18 μg L-1, respectively) but was less sensitive compared to growth for S. capricornutum (48-h and 72-h EC50 values of 4.9 and 7.5 μg L-1, respectively). For the other five species tested, inhibition of FDA fluorescence was relatively insensitive to copper, even at copper concentrations that severely inhibited cell division rate. These short-term bioassays that detect sublethal endpoints may provide a more rapid and cost-effective way of monitoring contaminant impacts in natural waters.

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http://hdl.handle.net/10453/3678