Association between overexpression of cytochrome P450 genes and deltamethrin resistance in Rhipicephalus microplus

Cytochrome P450 monooxygenases mediated metabolic detoxification has been recognized as one of the mechanisms involved in resistance to pyrethroids, which is a class of pesticides that includes acaricides such as deltamethrin. Several cytochrome P450 (CYP) genes were identified in arthropod pests which are upregulated in response to exposure to pesticides used as acaricides. However, to date, limited information is available with respect to CYP genes and their response to acaricide exposure in ticks. We cloned and sequenced four CYP genes, the CYP41, CYP3006G8, CYP319A1 and CYP4W1 from reference susceptible IVRI-I strain of Rhipicephalus microplus. The expression pattern of the genes was investigated using qPCR in reference susceptible IVRI-I, pyrethroid-resistant IVRI-IV and multi-acaricide resistant IVRI-V strains. The effect of a single exposure of deltamethrin, at a concentration of 2600 mu g/mL and 299.7 mu g/mL on IVRI-IV and IVRI-V strains, respectively, on the expression of the four CYP genes was evaluated. In IVRI-IV strain, the CYP41 gene was highly overexpressed (FC 8.72) while CYP3006G8 was underexpressed with FC of 0.06. All the four genes were overexpressed in IVRI-V strain. After exposure to deltamethrin, the CYP3006G8 transcript levels were significantly upregulated at all time intervals in both resistant strains with the highest FC of 11.62 at 12 h in IVRI-IV and 13.38 at 3 h in IVRI-V. Our results suggest that the constitutive overexpression of CYP41 and deltamethrin induced upregulation of CYP3006G8 contribute to the development of pyrethroid resistance, specifically deltamethrin, in these two reference strains.