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Development of an improved rapid enzyme inhibition bioassay with marine and freshwater microalgae using flow cytometry.
Arch Environ Contam Toxicol. 2001 May; 40(4):469-80.AE

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 microg Cu L(-1) (95% confidence limits 88-143) and 51 microg Cu L(-1) (95% confidence limits 38-70) for S. capricornutum and 47 microg Cu L(-1) (95% confidence limits 43-51) and 9.1 microg 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 microg 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 microg 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.

Authors+Show Affiliations

Franklin NM
Centre for Advanced Analytical Chemistry, CSIRO Energy Technology, Bangor, New South Wales, Australia. natasha.franklin@det.csiro.au
Adams MS
No affiliation info available
Stauber JL
No affiliation info available
Lim RP
No affiliation info available

MeSH

Biological AssayCopperCost-Benefit AnalysisEnvironmental MonitoringEsterasesEukaryotaFlow CytometryFluoresceinsLethal Dose 50Xenobiotics

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

11525489

Citation

Franklin, N M., et al. "Development of an Improved Rapid Enzyme Inhibition Bioassay With Marine and Freshwater Microalgae Using Flow Cytometry." Archives of Environmental Contamination and Toxicology, vol. 40, no. 4, 2001, pp. 469-80.
Franklin NM, Adams MS, Stauber JL, et al. Development of an improved rapid enzyme inhibition bioassay with marine and freshwater microalgae using flow cytometry. Arch Environ Contam Toxicol. 2001;40(4):469-80.
Franklin, N. M., Adams, M. S., Stauber, J. L., & Lim, R. P. (2001). Development of an improved rapid enzyme inhibition bioassay with marine and freshwater microalgae using flow cytometry. Archives of Environmental Contamination and Toxicology, 40(4), 469-80.
Franklin NM, et al. Development of an Improved Rapid Enzyme Inhibition Bioassay With Marine and Freshwater Microalgae Using Flow Cytometry. Arch Environ Contam Toxicol. 2001;40(4):469-80. PubMed PMID: 11525489.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development of an improved rapid enzyme inhibition bioassay with marine and freshwater microalgae using flow cytometry. AU - Franklin,N M, AU - Adams,M S, AU - Stauber,J L, AU - Lim,R P, PY - 2001/8/30/pubmed PY - 2001/9/14/medline PY - 2001/8/30/entrez SP - 469 EP - 80 JF - Archives of environmental contamination and toxicology JO - Arch Environ Contam Toxicol VL - 40 IS - 4 N2 - 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 microg Cu L(-1) (95% confidence limits 88-143) and 51 microg Cu L(-1) (95% confidence limits 38-70) for S. capricornutum and 47 microg Cu L(-1) (95% confidence limits 43-51) and 9.1 microg 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 microg 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 microg 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. SN - 0090-4341 UR - https://www.unboundmedicine.com/medline/citation/11525489/Development_of_an_improved_rapid_enzyme_inhibition_bioassay_with_marine_and_freshwater_microalgae_using_flow_cytometry_ DB - PRIME DP - Unbound Medicine ER -