The oil industry is a major source of contamination in Peru, and wastewater and sediments containing oil include harmful substances that may have acute and chronic effects. This study determined polycyclic aromatic hydrocarbon (PAH) concentrations by GC/MS, mutagenicity using TA98 and TA100 bacterial strains with and without metabolic activation in the Muta-ChromoPlate™ test, and Microtox® 5-min EC50 values of Peruvian crude oil, and water and sediment pore water from the vicinity of San José de Saramuro on the Marañón River and Villa Trompeteros on the Corrientes River in Loreto, Peru. The highest total PAH concentration in both areas was found in water (Saramuro = 210.15 μg/ml, Trompeteros = 204.66 μg/ml). Total PAH concentrations in water from San José de Saramuro ranged from 9.90 to 210.15 μg/ml (mean = 66.48 μg/ml), while sediment pore water concentrations ranged from 2.19 to 70.41 μg/ml (mean = 24.33 μg/ml). All water samples tested from Saramuro and Trompeteros sites, and one out of four sediment pore water samples from Trompeteros, were found to be mutagenic (P < 0.001). One sediment pore water sample in Saramuro was determined to have a measurable toxicity (Microtox EC50 = 335.1 mg/l), and in Trompeteros, the EC50 in water and sediment pore water ranged from 25.67 to 133.86 mg/l. Peruvian crude oil was mutagenic using the TA98 strain with metabolic activation, and the EC50 was 17.18 mg/l. The two areas sampled had very high PAH concentrations that were most likely associated with oil activities, but did not lead to acute toxic effects. However, since most of the samples were mutagenic, it is thought that there is a greater potential for chronic effects.

Induction of the cytochrome P450 enzymes in fish, measured as ethoxyresorufin-O-deethylase (EROD) activity, has been extensively used as a biomarker in assessing exposures and responses of aquatic organisms to contaminants. This study focused on whether exposure to Troy (Alabama) wastewater treatment plant (TWWTP) effluent at the Walnut Creek mixing zone, induced transcription of mRNA for cytochrome P4501A1 enzyme production and increased EROD activity in channel catfish (Ictalurus punctatus). Water from Big Creek (Barbour County, AL), regarded as least impacted by pollutants, was used as a reference source for administration of a positive control chemical (PCB – Aroclor 1254). All water samples were transported from field sites to the Troy University laboratory for conducting the study. Reverse transcriptase-polymerase chain reaction (RT-PCR) indicated the presence of compounds capable of inducing transcription of CYP1A1 mRNA in catfish liver. Increased transcription of mRNA for cytochrome P4501A1 enzyme production, confirmed by a 3-fold induction of EROD activity, was found in catfish exposed to water from the TWWTP mixing zone on Walnut Creek compared to catfish exposed to Walnut Creek Upstream. Catfish exposed to water from Big Creek treated with PCBs were found to have only slightly higher enzyme activities than those exposed to water from Big Creek Control, but a 2.7-fold EROD level was found in catfish exposed to Big Creek Control compared to Walnut Creek Upstream. Determining the induction of cytochrome P450 and subsequent enzymatic activity in catfish and other fish species common to this region could be useful early molecular warnings of possible pollution effects, beyond those methods typically used to characterize water quality.

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The objectives of this study were as follows: 1) to establish a baseline ethoxyresorufin-O-deethylase (EROD) activity level in channel catfish (Ictalurus punctatus), 2) to assess changes in induction of cytochrome P450 enzyme in channel catfish following exposure to creek water at the discharge point from the Troy (Alabama) Wastewater Treatment Plant (TWWTP) compared to upstream samples from Walnut Creek, 3) to compare EROD activity in populations maintained in laboratory and field settings, and 4) to quantify cytochrome P450 gene expression. Enzyme activity was measured fluorometrically and CYP1 gene expression was analyzed by quantitative real-time reverse transcription polymerase chain reaction. A mean EROD baseline was established at 0.03 nmol/min/µg of protein. The overall mean field effluent (TF) EROD had a significant 5-fold increase over field upstream (UF) exposed catfish; and overall mean laboratory effluent (TL) exposed catfish EROD had a significant 1.8-fold increase over laboratory upstream (UL) exposed catfish. Field exposures generally showed more robust enzyme induction over laboratory exposures on all sampling days. Ex- pression of the CYP1B gene following TF exposure was 6-fold over UF. Results suggested that in situ exposure to wastewater pollutants using caged test organisms provided a much more sensitive local monitor of pollutant exposure and biological impact than ex situ toxicological studies