Cytochrome P450 and dependent activities in unexposed and PAH-exposed terrestrial annelids

Toxicity, bioaccumulation and metabolism of pesticides in the earthworm

“…of 54, 56 and 58 kDa were identified by SDS-PAGE; the smallest one weakly reacted with the polyclonal antibody to perch hepatic CYP1A in western blotting but with trace EROD activity observed. 114) The largest isoform was separately reported in the same species together with AE and ODM activities. 115) L. terrestris was found to possess at least three CYP isoforms (48, 51 and 53 kDa) only with BROD activity, none of which reacted with the polyclonal antibodies against intact rat liver CYPs or those induced by 3-methylcholanthrene (3MC) or phenobarbital (PB).…”

“…117,122,124,125) However, the difference spectrum of the reduced CO-treated microsomal fraction showed the peak at 420 nm instead of the 450 nm peak typical for CYPs, most likely due to the interference by giant hemoglobin characteristic to the earthworms. 114,122,123,129) The presence of CYP in the microsomal fraction was spectrophotometrically supported after removal of the hemoglobin through successive chromatographic purifications. 114,123,124) The estimated CYP content in E. fetida, 114) L. terrestris 123,124) and Dendrobaena veneta 130) was 0.008-0.16 nmol/mg protein, depending on the purification level and the experimental conditions in the enzyme kinetic assays.…”

“…114,122,123,129) The presence of CYP in the microsomal fraction was spectrophotometrically supported after removal of the hemoglobin through successive chromatographic purifications. 114,123,124) The estimated CYP content in E. fetida, 114) L. terrestris 123,124) and Dendrobaena veneta 130) was 0.008-0.16 nmol/mg protein, depending on the purification level and the experimental conditions in the enzyme kinetic assays. Many studies show that both the CYP content and its activity are lower than those in rats and fish.…”

Earthworm Biomarkers as Tools for Soil Pollution Assessment

“…However, induction of CYP1A in earthworms has demonstrated to be quite difficult to be measured because of interference from endogenous pigments (Liimatainen & Hänninen, 1982) and the identification of non-inducible forms of cytochrome P450 (Milligan et al, 1986). Achazi et al (1998), by utilizing ethoxy-, pentoxy-and benzoxyresorufin as substrates for monooxygenase activity, demonstrated pentoxy-resorufin-Odealkylase (PentROD) and BenzROD activities in Eisenia fetida microsomes, but exposure of the animals for up to four weeks to 100 mg fluoranthene or benzo[a]pyrene kg-1 soil (dry weight) did not induce significant changes in the activity of these monooxygenases. The same authors demonstrated the presence of etoxy-resorufin-Odealkylase (EROD) and PentROD activities in Eisenia crypticus but failed to demonstrate an induction of these activities following xenobiotic exposure.…”

“…On the other hand short-term exposure to benzo[a]pyrene by feeding reduced the EROD activity significantly by 45%, but did not affect PentROD activity. After long-term (8 weeks) exposure to benzo[a]pyrene in the agar-agar medium EROD activity was not changed but PentROD was decreased to zero (Achazi et al, 1998). Glutathione transferases (GSTs) form a ubiquitous superfamily of multi-functional dimeric enzymes (w50 kDa) with roles in phase-II detoxification.…”