Biochemical diagnosis of organophosphate-insensitivity with neural acetylcholinesterase extracted by sonication from the adult tick synganglion

A sonication method for the homogeneous extraction of acetylcholinesterase (AChE) from the synganglia of adult ticks is described. The method provides for the extraction of sufficient AChE for multiple assays of enzyme activity in the presence of specific organophosphate (OP) inhibitors for the rapid diagnosis of OP-insensitivity and assignment of homozygous susceptible (SS), heterozygous resistant (RS), and homozygous resistant (RR) genotypes to individual ticks. A single synganglion from adult ticks of either gender and various stages of feeding can successfully be used for AChE extraction. The study presents the results of diagnostic screening of four Boophilus microplus strains for OP-insensitivity. The extraction method and these findings should find utility in support of researchers involved in the mitigation of acaricide resistance in tick populations worldwide, and in particular, the Cattle Fever Tick Surveillance and Quarantine Program maintained by USDA-APHIS/Veterinary Services along the south Texas border that prevents reentry of Boophilus spp. into the United States from endemic populations in Mexico.