Brown trout as a sentinel organism for organic pollution in the field using catalytic and immunochemical assays of cytochrome P-450 1A

The measurement in some living organisms of adequate biomarkers (e.g. cytochrome P-450) to assess the organic pollution in freshwater ecosystems is well established. However, the sensitivity of this approach depends on the analytical measurement method employed and on the chosen living organism for the biomonitoring. Three analytical methods were compared for measuring cytochrome P-450 1A levels in wild brown trout (Salmo trutta): a catalytic one, based on measurement of the ethoxyresorufin-O-deethylase (EROD) activity, and two immunochemical methods, namely, enzyme-linked immunosorbent assay (ELISA) and western blotting. The P-450 1A levels in those animals from a river located in an industrialized area (Trubia River, Northern Spain) and also from individuals living in a low-contamination reference area have been studied. Significant differences (Mann-Whitney U-test, P < 0.01) between rivers were found (ELISA and EROD assays), with the values for Trubia river being similar to those observed in laboratory experiments with well known P-450 1A inducers. However, no significant differences were observed in terms of sex and age. Western blot analysis confirmed the presence of a single band of 56 kDa (corresponding with P-450 1 A protein) in microsomes of fish caught in the Trubia river. On the other hand, and associated with the chemical analysis of PAHs in the waters of both rivers by SPME-GC-MS, high levels of naphthalene (P-450 1A inducer) in the contaminated river were found. In brief, a wide difference between basal levels and P-450 1A induction levels could be detected in trout living in natural field conditions using both EROD activity assay and immunochemical methods. Therefore, brown trout could constitute a good sentinel organism to biomonitor the exposure to PAHs in rivers using P-450 1A measurements.