Use of a highly sensitive recombinant hepatoma cell method to determine dioxin concentrations in samples of fish and crab from a hotspot area

Abstract

A new and easy fast-screening test (the Ad-DR (adenoviral vector-dioxin response) bioassay) for dioxins in biological samples from highly dioxin-contaminated areas was developed. The aryl-hydrocarbon-receptor (AhR) reporter system was utilized to transport a dioxin-responsive-element (DRE) via an adenovirus vector into rat hepatoma (H4IIE) cells before each experiment; these DRE-H4IIE cells were utilized in the Ad-DR bioassay. Biological extracts were simultaneously analyzed by the Ad-DR bioassay and high resolution gas chromatography/high resolution mass spectrometry (HRGC/HRMS). A good correlation was found between the results of the HRGC/HRMS assay and those of the Ad-DR bioassay (R(2) = 0.920, p < 0.001). The bio-analytical equivalent (BEQ) value found in fish or crab caught in the abandoned pentachlorophenol plant (AP) was extremely high compared with the BEQ in fish or crab caught in two rivers nearby this abandoned plant. Dioxins were more heavily bioaccumulated in fish viscera than in fish muscles or in the whole fish. Two-way analysis of variance tests identified the significant effects of fish collection site, fish or crab tissue sample and the interaction between them on dioxin levels in the tissues of these aquatic animals. In conclusion, the Ad-DR bioassay is a useful tool to determine dioxin levels in samples of fish and crab. Compared with fish tissues, where a sample is taken (in the PCP plant or nearby rivers) is the most important factor to determine bioaccumulation of dioxins in fish.

Polychlorinated Dibenzo-p-dioxin and Dibenzofuran (PCDD/F) Emission Behavior during Incineration of Laboratory Waste. Part 1: Emission Profiles Obtained Using Chemical Assay and Bioassay

“…Therefore, a highly sensitive recombinant hepatoma cell method (adenoviral vector-dioxin response (Ad-DR) bioassay), which takes much less time, was also adopted to analyze the PCDD/F concentration of specimens. Ad-DR bioassay has been demonstrated as a possible screening tool and has been shown to produce results consistent with those of high-resolution gas chromatography/ high-resolution mass spectrometry (HRGC/HRMS) (Chao et al, 2010(Chao et al, , 2012Lin et al, 2013). This study confirms the feasibility of this quick method for ash samples.…”

“…According to the linear regressions, the BEQ can serve as an index of PCDD/F emission to quickly predict the approximate PCDD/F concentration for HRGC/HRMS analysis. In addition, the PCDD/F distributions for I-TEQs and BEQs have similar profiles (Chao et al 2010(Chao et al , 2012Lin et al, 2013). Therefore, Ad-DR bioassay may be a promising method for investigating PCDD/F emission behavior with reduced time and the cost.…”

“…The dioxin bioassay (Ad-DR bioassay) used rat hepatoma cells (H4IIE cells) infected with AdEasy virus expressing TATA-4 × DRE-Luc (a luciferase reporter gene driven by a promoter with four copies of dioxin-responsive element (DRE)). Ad-DR bioassay with the virus construct has been proven to be useful for the quantitative analysis of dioxins and dioxin-like compounds (Lin et al, 2013). Details of the bioassay procedure can be found elsewhere (Lin et al, 2013).…”

“…Ad-DR bioassay with the virus construct has been proven to be useful for the quantitative analysis of dioxins and dioxin-like compounds (Lin et al, 2013). Details of the bioassay procedure can be found elsewhere (Lin et al, 2013). The luciferase activity was expressed as relative light units (RLUs)/well.…”

Combination of a Fast Cleanup Procedure and a DR-CALUX® Bioassay for Dioxin Surveillance in Taiwanese Soils

“…The quality assurance/quality control (QA/QC) of this effective extraction and cleanup system meets the criteria in U.S. EPA Method 1613B, 1668A, 1614, 4425, and 4435 as well as the Taiwanese EPA Method [13]. The fast extraction approach coupled with cleanup processes have been applied to various matrices, including soils, sediments, and fish [25,26,40]. …”