Fluoroquinolone antibiotics and organophosphate pesticides induce carbonylation on Eisenia fetida muscle proteins

Abstract

Protein carbonylation is an irreversible oxidative modification that has been associated with a decrease in the quality and nutritional value of products of animal origin. Generally, the carbonylation is attributed to processes of slaughter, processing, and cold storage of products. However, in vitro studies have shown that fluoroquinolone and organophosphate pesticides residues at their maximum residue limits (MRL) can promote carbonylation of animal proteins. Though, this effect on in vivo conditions has not yet been evaluated. Thus, Eisenia fetida was chosen as a model to assay their oxidant effect. For this, adult earthworms were exposed to artificial soil contaminated with ciprofloxacin, danofloxacin, fenthion, and diazinon at three concentrations (0.5, 1.0, and 1.5 MRL) for 28 days. Then, these were purged and sacrificed to obtain the muscle region between the anus and preclitellum. The muscle samples were cold macerated to obtain muscle proteins, which were used for protein quantification, determination of carbonyl levels, and carbonyl protein profiles employing Bradford, Dot-blot, and Western blot assays, respectively. The results showed that at each concentration assayed, all pollutants induced significant carbonylation respect to control (p < 0.05). Additionally, mass spectrometry-based analysis (MALDI-TOF/TOF) identified actin as the protein most susceptible to carbonylation promoted by these substances. Therefore, these findings show for the first time the oxidant power of fluoroquinolones and organophosphates pesticides at MRLs concentrations on muscle proteins under in vivo conditions. Fact causes concern due to the homology of muscle proteins in eukaryotes, which allow to hypothesize that this effect could also be experienced by proteins from food-producing animals in the same way that observed in in vitro studies.