Aryl Hydrocarbon Receptor-Mediated Toxic Potency of Dissolved Lipophilic Organic Contaminants Collected From Lincoln Creek, Milwaukee, Wisconsin, Usa, to PLHC-1 (Poeciliopsis Lucida) Fish Hepatoma Cells

Abstract

Abstract-LincolnCreek is a severely degraded urban stream located in Milwaukee County, Wisconsin, USA. As part of a comprehensive study on effects of urban storm water runoff on the stream biota, an in vitro bioassay with PLHC-1 (Poeciliopsis lucida) fish hepatoma cells was used to assess potential toxic potency of aryl hydrocarbon receptor (AhR)-active compounds, collected by semipermeable membrane devices (SPMDs) exposed to Lincoln Creek water. Dialysates from SPMDs exposed to Lincoln Creek water caused marked cytochrome P4501A induction in PLHC-1. Toxic potency of dialysates, expressed as bioassayderived 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TCDD-EQ) ranged from 1,300 to 6,600 pg TCDD-EQ/g SPMD for 14-d exposures. Dialysates from SPMDs exposed to stream water at base flow had potencies consistently lower than those exposed to storm-flow (high-flow) events that occurred during the same 14-d period. Polychlorinated biphenyls were not detectable in the dialysates. Gas chromatography-mass spectrometry analysis identified polycyclic aromatic hydrocarbons (PAHs) as major contaminants in the dialysates. A log-log correlation of total PAHs and TCDD-EQ yielded an r 2 of 0.802. Empirical evidence suggests that AhR-active PAHs can account for about 20 to 50% of the potency observed.

The Use of Semipermeable Membrane Devices to Concentrate Chemicals in Oil Refinery Effluent on the Mackenzie River

“…the oil sands wastewater-exposed SPMDs. SPMDs deployed for 14 days in Lincoln Creek, Milwaukee, which receives PAH-contaminated stormwater runoff, had 1300-6600 pg TCDD-EQ/g SPMD (Villeneuve et al, 1997), a potency very similar to those of our SPMDs from Norman Wells refinery effluent.…”

“…Such samples , 1996). SPMDs have been used to concentrate MFOinducing chemicals from pulp mill effluents; from oil sands, mining, and refining wastewaters; and from river and stream waters naturally high in PAHs (Parrott et al, 1996a,b;Villeneuve et al, 1997).…”

Integrated Biomonitoring of Dioxin-like Compounds for Waste Management and Environment.

“…In vitro micro-EROD assays have been simplified using culture cells such as H4IIE rat hepatoma cell line, mouse Hepa lclc7 cell line, the PLHC-1 fish hepatoma cell line, rainbow trout liver cell line RTL-W1 and human hepatoma HepG2 cells in 96-well microplates for the purpose more related to analysis of PHAHs [9][10][11][12][13][14][15][16][17][18][19] . For in vitro assay, induction of EROD activity caused by test samples can be compared with the induction curve of TCDD standard to determine biological TEQ (EROD-TEQ) values in samples.…”

Mechanisms of action for aryl hydrocarbon receptor ligands in the PLHC-1 cell line

“…These cells have been used for investigations of cytotoxicity (Babich et al 1991b;Ryan and Hightower 1994;Bruschweiler et al 1995;Huuskonen et al 1998a;Fent and Batscher 2000) and effects of AHR ligands (Bruschweiler et al I996a;Bruschweiler et al 1996b;Celander et al 1996;Celander et al 1997;Villeneuve et al 1997;Huuskonen et al 1998b;Huuskonen et al 1998c;Smeets et al 1999;Huuskonen et al 2000). PLHC-I cells show induction of CYPIA (an AHR-mediated response) in response to AHR agonists (Hahn et al 1993;Hahn et al 1996a;Hahn et al 1996b;Hestermann et al in press).…”