Benzo[a]pyrene cytotoxicity in right whale (Eubalaena glacialis) skin, testis and lung cell lines

Godard, Céline A. J.; Wise, Sandra S.; Kelly, Roxanne; Goodale, Britton C.; Kraus, Scott D.; Romano, Tracy A.; O’Hara, Todd M.; Wise, John Pierce

doi:10.1016/j.marenvres.2006.04.016

Cited by 24 publications

(9 citation statements)

References 10 publications

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“…Most studies on cell cultures of wild cetaceans have thus targeted fibroblasts. In previous studies, cetacean fibroblast cells have been isolated from skin, ,− lungs, testes, muscle, and cartilage . This study proposed a non-invasive and efficient cell culture method using the flipper tip of a stranded cetacean.…”

Section: Resultsmentioning

confidence: 99%

In Vitro Cytotoxicity and Risk Assessments of Environmental Pollutants Using Fibroblasts of a Stranded Finless Porpoise (Neophocaena asiaeorientalis)

Ochiai

Kurihara

Hirano

et al. 2020

Environ. Sci. Technol.

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Cetaceans accumulate high levels of environmental pollutants, yet their toxicological studies have been difficult due to technical and ethical issues. It is essential to identify and fill the current knowledge gaps in the in vitro assays available for cetaceans. The present study establishes a novel in vitro assay that uses the fibroblasts of a finless porpoise (Neophocaena asiaeorientalis) (FF) stranded in the Seto Inland Sea (SIS) to answer questions about the cytotoxicity and risks of environmental pollutants. FF were treated with 17 compounds including polychlorinated biphenyls (PCBs) and dichlorodiphenyltrichloroethane and their metabolites (DDTs) and evaluated for cytotoxicity, viability, and apoptosis. The results of FF were compared with those of human fibroblasts (HF). The relative potencies of the test compounds were comparable between the two species, as EC50 of these compounds significantly correlated for FF and HF. Exposure-activity ratios (EARs) revealed that accumulation of PCBs and DDTs are likely to pose adverse effects at the cellular level in the SIS finless porpoises, as their tissue concentrations exceeded EC50 values obtained in this study. This study successfully evaluated the risks of environmental pollutants using cetacean fibroblasts isolated by a non-invasive method that may be applied to various cetacean species and compounds.

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Section: Resultsmentioning

confidence: 99%

In Vitro Cytotoxicity and Risk Assessments of Environmental Pollutants Using Fibroblasts of a Stranded Finless Porpoise (Neophocaena asiaeorientalis)

Ochiai

Kurihara

Hirano

et al. 2020

Environ. Sci. Technol.

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“…There is a critical need to reliably assess and monitor the effects of environmental pollutants on sea turtle populations within the legal constraints of their threatened or endangered status. Traditional invasive or lethal toxicology testing methods are very seldom suitable to endangered species research. , For this reason, toxicology research in sea turtles so far has focused mainly on contaminant burden analyses in tissues, − with rare forays into animal dosing , or egg exposure studies. , Immortal cell lines were developed for virology in the green sea turtle (Chelonia mydas) almost exclusively from animals affected by fibropapillomatosis − or from embryos . Pioneering in vitro toxicity testing in sea turtles used these cell lines to examine the effect of organochlorines on cytochrome P450 aromatase and test metal sensitivity. , Primary skin fibroblasts and embryonic cultures were recently established and partially characterized in the hawksbill sea turtle (Eretmochelys imbricata) and olive ridley sea turtle (Lepidochelys olivacea). − Primary skin fibroblasts from one hawksbill sea turtle were subsequently evaluated for chromium toxicity and a preliminary assessment of thermal effects on heat shock protein expression was conducted in embryonic cultures in the same species .…”

Section: Introductionmentioning

confidence: 99%

Establishment, Characterization, and Toxicological Application of Loggerhead Sea Turtle (Caretta caretta) Primary Skin Fibroblast Cell Cultures

Webb

Zychowski

Bauman

et al. 2014

Environ. Sci. Technol.

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Pollution is a well-known threat to sea turtles but its impact is poorly understood. In vitro toxicity testing presents a promising avenue to assess and monitor the effects of environmental pollutants in these animals within the legal constraints of their endangered status. Reptilian cell cultures are rare and, in sea turtles, largely derived from animals affected by tumors. Here we describe the full characterization of primary skin fibroblast cell cultures derived from biopsies of multiple healthy loggerhead sea turtles (Caretta caretta), and the subsequent optimization of traditional in vitro toxicity assays to reptilian cells. Characterization included validating fibroblast cells by morphology and immunocytochemistry, and optimizing culture conditions by use of growth curve assays with a fractional factorial experimental design. Two cell viability assays, MTT and lactate dehydrogenase (LDH), and an assay measuring cytochrome P4501A (CYP1A) expression by quantitative PCR were optimized in the characterized cells. MTT and LDH assays confirmed cytotoxicity of perfluorooctanoic acid at 500 μM following 72 and 96 h exposures while CYP1A5 induction was detected after 72 h exposure to 0.1-10 μM benzo[a]pyrene. This research demonstrates the validity of in vitro toxicity testing in sea turtles and highlights the need to optimize mammalian assays to reptilian cells.

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“…CYP1A induction is a recognized and widely used measure of exposure to, and molecular effects of, oil and PAHs in humans, laboratory animals and wildlife species (Godard et al, 2004(Godard et al, , 2006Stegeman et al, 1992;Webb et al, 2014) including birds (Brausch et al, 2010;Brunström et al, 1991, Custer et al, 2000Lee et al, 1985;Lee et al, 1986;Peakall et al, 1989,). Avian species express two forms of CYP1A: CYP1A4 and CYP1A5 (Gilday et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

Reprint of: CYP1A protein expression and catalytic activity in double-crested cormorants experimentally exposed to Deepwater Horizon Mississippi Canyon 252 oil

Alexander

Hooper

Cacela

et al. 2017

Ecotoxicology and Environmental Safety

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Double-crested cormorants (Phalacrocorax auritus, DCCO) were orally exposed to Deepwater Horizon Mississippi Canyon 252 (DWH) oil to investigate oil-induced toxicological impacts. Livers were collected for multiple analyses including cytochrome P4501A (CYP1A) enzymatic activity and protein expression. CYP1A enzymatic activity was measured by alkoxyresorufin O-dealkylase (AROD) assays. Activities specific to the O-dealkylation of four resorufin ethers are reported: benzyloxyresorufin O-debenzylase (BROD), ethoxyresorufin O-deethylase (EROD), methoxyresorufin O-demethylase (MROD), and pentoxyresorufin O-depentylase (PROD). CYP1A protein expression was measured by western blot analysis with a CYP1A1 mouse monoclonal antibody. In study 1, hepatic BROD, EROD, and PROD activities were significantly induced in DCCO orally exposed to 20ml/kg body weight (bw) oil as a single dose or daily for 5 days. Western blot analysis revealed hepatic CYP1A protein induction in both treatment groups. In study 2 (5ml/kg bw oil or 10ml/kg bw oil, 21day exposure), all four hepatic ARODs were significantly induced. Western blots showed an increase in hepatic CYP1A expression in both treatment groups with a significant induction in birds exposed to 10ml/kg oil. Significant correlations were detected among all 4 AROD activities in both studies and between CYP1A protein expression and both MROD and PROD activities in study 2. EROD activity was highest for both treatment groups in both studies while BROD activity had the greatest fold-induction. While PROD activity values were consistently low, the fold-induction was high, usually 2nd highest to BROD activity. The observed induced AROD profiles detected in the present studies suggest both CYP1A4/1A5 DCCO isoforms are being induced after MC252 oil ingestion. A review of the literature on avian CYP1A AROD activity levels and protein expression after exposure to CYP1A inducers highlights the need for species-specific studies to accurately evaluate avian exposure to oil.

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Benzo[a]pyrene cytotoxicity in right whale (Eubalaena glacialis) skin, testis and lung cell lines

Cited by 24 publications

References 10 publications

In Vitro Cytotoxicity and Risk Assessments of Environmental Pollutants Using Fibroblasts of a Stranded Finless Porpoise (Neophocaena asiaeorientalis)

In Vitro Cytotoxicity and Risk Assessments of Environmental Pollutants Using Fibroblasts of a Stranded Finless Porpoise (Neophocaena asiaeorientalis)

Establishment, Characterization, and Toxicological Application of Loggerhead Sea Turtle (Caretta caretta) Primary Skin Fibroblast Cell Cultures

Reprint of: CYP1A protein expression and catalytic activity in double-crested cormorants experimentally exposed to Deepwater Horizon Mississippi Canyon 252 oil

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