Gills are major targets for acute metal toxicity in fish, due to their permanent contact with aquatic pollutants. To assess the effects of metals on gills of the Leuciscus cephalus (chub), fish individuals were collected from two sites in the Tur River, Romania, in upstream (site 1) and downstream (site 2) of a metal pollution source. Quantitative and hyperspectral analyses showed that Zn, Sr, and Fe concentrations were significantly higher in gills from site 2 compared with site 1. Malondialdehyde and advanced oxidation protein products levels increased 17 and 28%, respectively, whereas reduced glutathione level diminished significantly in the gills of fish collected from site 2 compared to site 1. The activities of superoxide dismutase, catalase, and glutathione-S-transferase increased significantly at 41, 21, and 28%, respectively. Proliferating cell nuclear antigen (PCNA) protein levels, as well as the amount of DNA damage, were significantly increased for site 2 compared with site 1. The induced oxidative stress generated hyperplasia, hypertrophy, and inflammation in the epithelial cells and apoptosis. Hence, this could suggest that gill cells have tried to counteract the oxidative stress-induced DNA fragmentation by PCNA up-regulation, but the PCNA expression decreased on longer time due to the low level of GSH, resulting in apoptosis.

This study assessed the distribution of heavy metals in the gills, kidney, and liver, correlated with the severity of histopathological changes, of three fish species with different feeding habitats (Barbus barbus, Squalius cephalus, and Chondrostoma nasus) from the Crișul Negru river, Romania. The levels of copper (Cu), chromium (Cr), cadmium (Cd), lead (Pb), and zinc (Zn) in fish tissues were measured by atomic absorption spectrophotometry. Histopathology and the expressions of TNF-α and proliferation cell nuclear antigen (PCNA) were investigated by immunohistochemistry and Western blot. Our data suggest a significant correlation between the bioconcentration level of metals and structural changes. The carnivorous species was the most affected compared to the omnivorous and herbivorous ones, and the most affected organ was the kidney. Moreover, the correlation of tissue damage with the PCNA and TNF-α expression levels revealed that the herbivorous species presented less extended lesions, likely due to higher activated repair mechanisms and lower levels of inflammation. In conclusion, our data and the subsequent statistical analysis suggest that feeding behavior could be correlated with the histopathological alterations and might be used for a more profound evaluation of aquatic environment safety and analysis of aquatic ecosystems.

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