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Title: Biomonitoring of aquatic sediments from Estonia with a fish liver cell line (PLHC-1)

Abstract

Sediments were collected from lake Vorstjaerv and River Narva, Estonia, and extracted with dichloromethane. PLHC-1 cells were exposed to the extracts of the dry sediments. Cytotoxicity (total protein content) and CYP1A inducibility (EROD activity) was detected in the exposed cells. The contents of selected PAHs in the R. Narva sediments were also analyzed. The sediments collected from R. Narva were more toxic and caused higher EROD induction in the cells than the sediments from L. Vortsjaerv. The sediments collected from the northern part of the L. Vortsjaerv showed cytotoxicity. The sediments collected from the southern part of L. Vortsjaerv showed slight induction in EROD activity. In R. Narva, the most toxic sediments were collected downstream from the city of Narva and the Baltic Thermal Power Plant. At sublethal level of the sediment concentrations, also the highest EROD activities were seen at these downstream sites. At the sites heavily polluted with PAHs, also the EROD activities were high. The correlation between the PAH contents and EROD activities was not so evident when the sediments contained less PAHs. The PLHC-1 cells could serve as a sensitive method for screening cytotoxic and enzyme-inducing effects of the sediment extracts. This helps in evaluating themore » potential hazard of the compounds bound in the sediments for aquatic life. Further, the PLHC-1 cells can be used in comparing the properties of the sediments collected from different lakes and rivers.« less

Authors:
; ; ;  [1];
  1. Univ. of Kuopio (Finland). Dept. of Physiology
Publication Date:
OSTI Identifier:
452044
Report Number(s):
CONF-961149-
Journal ID: ISSN 1087-8939; TRN: IM9715%%76
Resource Type:
Conference
Resource Relation:
Conference: 17. annual meeting of the Society of Environmental Toxicology and Chemistry: partnerships for the environment - science, education, and policy, Washington, DC (United States), 17-21 Nov 1996; Other Information: PBD: 1995; Related Information: Is Part Of SETAC 17. annual meeting -- Abstract book. Partnerships for the environment: Science, education, and policy; PB: 378 p.
Country of Publication:
United States
Language:
English
Subject:
54 ENVIRONMENTAL SCIENCES; 56 BIOLOGY AND MEDICINE, APPLIED STUDIES; BIOLOGICAL INDICATORS; WATER POLLUTION; POLYCYCLIC AROMATIC HYDROCARBONS; ESTONIA; FISHES; MONITORING; OXIDOREDUCTASES; ENZYME ACTIVITY; BIOLOGICAL MARKERS

Citation Formats

Huuskonen, S., Ritola, O., Koponen, K., Lindstroem-Seppae, P., and Tuvikene, A. Biomonitoring of aquatic sediments from Estonia with a fish liver cell line (PLHC-1). United States: N. p., 1995. Web.
Huuskonen, S., Ritola, O., Koponen, K., Lindstroem-Seppae, P., & Tuvikene, A. Biomonitoring of aquatic sediments from Estonia with a fish liver cell line (PLHC-1). United States.
Huuskonen, S., Ritola, O., Koponen, K., Lindstroem-Seppae, P., and Tuvikene, A. Sun . "Biomonitoring of aquatic sediments from Estonia with a fish liver cell line (PLHC-1)". United States. doi:.
@article{osti_452044,
title = {Biomonitoring of aquatic sediments from Estonia with a fish liver cell line (PLHC-1)},
author = {Huuskonen, S. and Ritola, O. and Koponen, K. and Lindstroem-Seppae, P. and Tuvikene, A.},
abstractNote = {Sediments were collected from lake Vorstjaerv and River Narva, Estonia, and extracted with dichloromethane. PLHC-1 cells were exposed to the extracts of the dry sediments. Cytotoxicity (total protein content) and CYP1A inducibility (EROD activity) was detected in the exposed cells. The contents of selected PAHs in the R. Narva sediments were also analyzed. The sediments collected from R. Narva were more toxic and caused higher EROD induction in the cells than the sediments from L. Vortsjaerv. The sediments collected from the northern part of the L. Vortsjaerv showed cytotoxicity. The sediments collected from the southern part of L. Vortsjaerv showed slight induction in EROD activity. In R. Narva, the most toxic sediments were collected downstream from the city of Narva and the Baltic Thermal Power Plant. At sublethal level of the sediment concentrations, also the highest EROD activities were seen at these downstream sites. At the sites heavily polluted with PAHs, also the EROD activities were high. The correlation between the PAH contents and EROD activities was not so evident when the sediments contained less PAHs. The PLHC-1 cells could serve as a sensitive method for screening cytotoxic and enzyme-inducing effects of the sediment extracts. This helps in evaluating the potential hazard of the compounds bound in the sediments for aquatic life. Further, the PLHC-1 cells can be used in comparing the properties of the sediments collected from different lakes and rivers.},
doi = {},
journal = {},
number = ,
volume = ,
place = {United States},
year = {Sun Dec 31 00:00:00 EST 1995},
month = {Sun Dec 31 00:00:00 EST 1995}
}

Conference:
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  • Planar halogenated hydrocarbons (PHHs) elicit dioxin-like effects in fish at extremely low doses. However, the relative potencies of PHHs in fish differ from mammals in both absolute magnitude and relative rank order. Therefore, the relative potencies of polychlorinated dibenzofurans (PCDFs) were examined in PLHC-1 fish hepatoma cells. The response of PHHs in this cell line correlates with their toxicity to cause early life stage toxicity in fish. Twenty PCDFs were selected by means of a multivariate chemical characterization in combination with a 2{sup 4}-factorial design and separated into a 10 congener training set and 10 congener validation set. Toxic equivalencymore » factors (TEFs) were determined for all 20 PCDF congeners based on ethoxyresorufin-O-deethylase (EROD) induction in the PLHC-1 cells. A quantitative structure-activity relation-ship (QSAR) model was constructed by partial least square modelling (PLS) of the physiochemical descriptor variables for the PCDFs in the training set and their EROD responses in the PLHC-1 cells. A predictive model was developed which accurately assessed the TEFs of the validation set of PCDFs. The model accounted for 87% (R{sup 2}) of the variance in TEFs and a cross-validated variance (Q{sup 2}) of 79%. From this QSAR, TEF values were predicted for all of the tetra through octa-chlorinated PCDFs in fish. Interestingly, there was a high degree of correlation among TEF values calculated in the PLHC-1 cells and the TEF values calculated in H411 E rat hepatoma cells.« less
  • Benzo(a)pyrene (BaP) can be transformed to 7,8 diol-9,10 epoxide BaP which is a major carcinogen. The cytochrome P45OIAI isoenzyme is playing an important role in this bioactivation. The coplanar PCB{number_sign}126 is a strong inducer of CYPIAI in vivo but also in the PLHC-1 fish cell line. This CYPIAI inducing property of PCB{number_sign}L26 could increase the BaP induced DNA-damage. In this study dose-response relationships for PCB{number_sign}126, BaP and their combination are presented, measured either as CYPIAI (EROD) induction or as DNA strand breaks. Strand breaks were measured by means of an alkaline unwinding assay. BaP is readily metabolized in this cellmore » line and induces strand breaks dose-dependently. PCB induces EROD in a dose-dependant way but does not cause DNA strand breaks significantly. PLHC-1 cells were pre-treated with PCB{number_sign}126 (LOEL,ED50 and maximal EROD induction) and then dosed with BaP (LOEL,ED5 and maximal strand break induction). DNA-strand breaks in the pre-treated cells were compared to cells treated with BaP alone. Results of these combined exposures will be presented and discussed.« less
  • The development of the multidrug resistance (MDR) phenotype in mammals is often mediated by the overexpression of the P-glycoprotein1 (Pgp, ABCB1) or multidrug resistance-associated protein (MRP)-like ABC transport proteins. A similar phenomenon has also been observed and considered as an important part of the multixenobiotic resistance (MXR) defence system in aquatic organisms. We have recently demonstrated the presence of ABC transporters in the widely used in vitro fish model, the PLHC-1 hepatoma cell line. In the present study we were able to select a highly resistant PLHC-1 sub-clone (PLHC-1/dox) by culturing the wild-type cells in the presence of 1 {mu}Mmore » doxorubicin. Using quantitative PCR a 42-fold higher expression of ABCB1 gene was determined in the PLHC-1/dox cells compared to non-selected wild-type cells (PLHC-1/wt). The efflux rates of model fluorescent Pgp1 substrates rhodamine 123 and calcein-AM were 3- to 4-fold higher in the PLHC-1/dox in comparison to the PLHC-1/wt cells. PLHC-1/dox were 45-fold more resistant to doxorubicin cytotoxicity than PLHC-1/wt. Similarly to mammalian cell lines, typical cross-resistance to cytotoxicity of other chemotherapeutics such as daunorubicin, vincristine, vinblastine, etoposide and colchicine, occurred. Furthermore, cyclosporine A, verapamil and PSC833, specific inhibitors of Pgp1 transport activity, completely reversed resistance of PLHC-1/dox cells to all tested drugs, resulting in EC50 values similar to the EC50 values found for PLHC-1/wt. In contrast, MK571, a specific inhibitor of MRP type of efflux transporters, sensitized PLHC-1/dox cells, neither to doxorubicin, nor to any other of the chemotherapeutics used in the study. These data demonstrate for the first time that a specific Pgp1-mediated doxorubicin resistance mechanism is present in the PLHC-1 fish hepatoma cell line. In addition, the fact that low micromolar concentrations of specific inhibitors may completely reverse a highly expressed doxorubicin resistance points to the fragility of Pgp1-mediated MXR defence mechanism in fish.« less
  • Lincoln Creek 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 bioassay-derived 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TCDD-EQ) ranged from 1,300 to 6,600 pg TCDD-EQ/g SPMD formore » 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{sup 2} of 0.802. Empirical evidence suggests that AhR-active PAHs can account for about 20 to 50% of the potency observed.« less
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