The biochemical factors responsible for parathion resistance in a ethyl fenitrothion-selected Yumenoshima I11 (EF-30) strain of the housefly were examined. Great difference (167-fold) in the Iso was observed between the resistant EF-30 (R) and susceptible SRS (S) strains in vitro, suggesting that altered acetylcholinesterase (AChE) in the housefly strain was an important factor in the resistance. The in vitro degradative activity of parathion and paraoxon in both strains was associated with the microsomal and soluble fractions and required NADPH and reduced glutahione (GSH), respectively. The R strain possessed higher activity for GSH S-transferase than the S strain, and this enzyme appears to be important in the resistance mechanism. The R strain was highly resistant to parathion (101,487-fold), but substitution of the methoxy group for ethoxy group decreased the resistance level (25,914-fold) and parathion could be a substrate of GSH S-transferase. It is concluded that the combination of some factors (altered AChE, and enhanced activity of cytochrome P450 dependent monooxygenase and GSH S-transferase) could be sufficient to account for the extremely high level of resistance to parathion and parathion-methyl, although a possible involvement of other factor(s) can not be excluded.