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Two‐dimensional gel analysis of stress proteins identified in Chironomus flaviplumus (Diptera: Chironomidae) exposed to 4‐nonylphenol

Toxicity of 4‐nonylphenol (4‐NP) in the Chironomidae Chironomus flaviplumus was analyzed using a proteomics approach that involved identifying proteins by two‐dimensional polyacrylamide gel electrophoresis (2‐D PAGE). Proteome analysis of 4‐NP‐treated samples on silver stained gels found alterations in the expression levels of three proteins compared with control samples. Hsp70 proteins, so‐called stress proteins, were studied in Chironomus flaviplumus exposed to different concentrations of 4‐nonylphenol (0, 30, 100, 150, 300 and 600 mg/L) in the laboratory and in the field in captured animals from site 1 (1 km from a chemical factory) and site 3 (16 km from a chemical factory). Hsp70 proteins were found in all samples tested, including controls, but differed in their expression levels. At more polluted sites (site 1), the samples treated with higher concentrations of 4‐NP more strongly expressed Hsp70. 2‐D spots were induced or enhanced in gels following injection of 4‐NP. Therefore, the induction of stress protein expression in Chironomus flaviplumus, in particular Hsp70, can be used as a biomarker for the evaluation of environmental conditions.