title: Occurrence of pharmaceutical products in a municipal effluent and toxicity to rainbow trout (Oncorhynchus mykiss) hepatocytes.
abstract: Pharmaceutical and personal care products (PPCPs) are found in municipal effluents and represent the major sources of contamination for the aquatic environment. A preliminary chemical analysis of wastewater identified several compounds associated with PPCPs, including caffeine, ibuprofen, naproxen, oxytetracycline, novobiocin, carbamazepine, gemfibrozil, bezafibrate, trimethoprim, sulfamethoxazole, and sulfapyridine. The purpose of this study was to examine the cytotoxic and oxidative effects of these products and other wastewater-related products (i.e., coprostanol, cotinine, estradiol-17beta, nonylphenol, and cholesterol) in primary cultures of rainbow trout hepatocytes. The redox activity of various PPCPs in trout (Oncorhynchus mykiss) liver microsomes was investigated in vitro by tracking the rate of oxidation of reduced nicotinamide adenine dinucleotide phosphate (NADPH) and the formation of lipid peroxidation (LPO) after a 60-min incubation period. In addition, primary cultures of rainbow trout hepatocytes were exposed to various drugs identified in the municipal effluent for 48 h at 15 degrees C. Our results show that most PPCPs (83%) accelerated the rate of NADPH oxidation in the presence of microsomes and 72% of them increased LPO in microsomal membranes. LPO levels were significantly correlated (R = 0.5; P<0.05) with the number of functional groups on the molecule's backbone (i.e., number of O, S, N, P/number of C and H) and negatively so (R = -0.44; P<0.05) with the octanol/water partition coefficient, suggesting that nucleophilicity and hydrophobicity are related to oxidative activity for these compounds. Exposure of trout hepatocytes to these products leads in many cases to decreased cell viability, increased CYP3A-related monooxygenase activity (benzylether resorufin dealkylase), and LPO. No induction of CYP1A1-related activity (7-ethoxyresorufin O-deethylase) was observed. Moreover, municipal effluent extracts (ethanol) were able to increase all the above responses in a dose-dependent manner. These results suggest that the basic redox properties of PPCPs could influence oxidative metabolism in liver cells and lead to oxidative damage. These products have the potential to produce a toxic response in aquatic organisms and the above biomarkers were shown to respond readily to PPCPs in aquatic organisms.
authors.affiliation: St. Lawrence Centre, Environment Canada, 105 McGill St., Montreal, Qc., Canada H2Y 2E7. francois.gagne@ec.gc.ca
authors.forename: F
authors.initials: F
authors.lastname: Gagné
authors.forename: C
authors.initials: C
authors.lastname: Blaise
authors.forename: C
authors.initials: C
authors.lastname: André
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003358:Cosmetics / Q000009:adverse effects* / Q000032:analysis / Q000737:chemistry; D051544:Cytochrome P-450 CYP3A / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D064420:Drug-Related Side Effects and Adverse Reactions; D022781:Hepatocytes / Q000187:drug effects* / Q000473:pathology; D007425:Intracellular Membranes / Q000187:drug effects / Q000378:metabolism; D015227:Lipid Peroxidation / Q000187:drug effects; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D000442:Octanols / Q000737:chemistry; D017686:Oncorhynchus mykiss; D018384:Oxidative Stress / Q000187:drug effects; D004364:Pharmaceutical Preparations / Q000032:analysis / Q000737:chemistry; D012722:Sewage / Q000009:adverse effects* / Q000032:analysis / Q000737:chemistry; D012995:Solubility; D018675:Toxicity Tests; D014867:Water / Q000737:chemistry; D014874:Water Pollutants, Chemical / Q000032:analysis / Q000633:toxicity*
chemical_list: D003358:Cosmetics; D000442:Octanols; D004364:Pharmaceutical Preparations; D012722:Sewage; D014874:Water Pollutants, Chemical; D014867:Water; D009249:NADP; D051544:Cytochrome P-450 CYP3A