Anti-cholinesterase resistance is in many cases caused by modified acetylcholinesterase (MACE). A comparison was made of toxicological data and AChE activity gathered from 21 field populations and nine laboratory strains of houseflies, Musca domestica L., to elucidate the best way of generating data to provide advice for management strategies and gathering information for resistance risk assessment on the organophosphates azamethiphos and dimethoate and the carbamate methomyl, which have been the primary insecticides used against adult houseflies in Denmark. Cluster analysis was performed and > 2000 houseflies were assigned to one of three phenotypes based on total acetylcholinesterase activity as well as inhibition by azamethiphos, methomyl or omethoate. A cluster, i.e. a phenotype, with high total AChE activity and high sensitivity to azamethiphos and less sensitivity to inhibition by methomyl and omethoate was shown to be linked to methomyl resistance. It was not possible to define any clusters that could be linked to azamethiphos or dimethoate resistance. The five mutations V180L, G262A, G262V, F327Y and G365A causing anticholinesterase resistance in houseflies were all identified in the Danish housefly strains. The data are very heterogeneous, and a correlation of molecular genetic background and resistance of phenotypes is not obvious with the available data.
Title: Resistance pattern and point mutations of insensitive acetylcholinesterase in a carbamate-resistant strain of housefly (Musca domestica) Liming T, Mingan S, Jiangzhong Y, Peijun Z, Chuanxi Z, Zhenhua T Ref: Pesticide Biochemistry and Physiology, 86:1, 2006 : PubMed
To examine the resistance pattern and point mutations present in the carbamate-resistant strain of housefly (SH-CBR), Musca domestica, from Shanghai, China, the flys resistance spectrum and ki ratios to organophosphate (OP) and carbamate (CB) insecticides were determined. The cDNA encoding acetylcholinesterase (AChE) from both a susceptible control strain (SH-S) and the SH-CBR strain of housefly were cloned and sequenced using RT-PCR. Based on two major patterns of target site resistance suggested by Russell et al. [R. J. Russell, C. Claudianos, P. M. Campbell, I. Horne, T. D. Sutherland, J. G. Oakeshott, Two major classes of target site insensitivity mutations confer resistance to organophosphate and carbamate, Pestic. Biochem. Physiol. 79 (2004), 8493.], the present results indicate a Pattern I resistance: resistance to CBs was greater than that to OPs; the SH-S/SH-CBR ki ratios were also greater for CBs than for OPs. The cDNA was 2079 nucleotides long, encoding a protein of 693 amino acids. The cDNA deduced amino acid sequence of the housefly had a high degree of identity with previously described insect AChEs and exhibited the same structural feature as vertebrate TcAChE. Three mutations in the region of the active site, V261L, G343A, and F408Y, which were identical to those identified in the 77M and YBOL housefly strains, were identified in the AChE from the SH-CBR strain. Additionally, a novel mutation D422L, was found at the outer surface of the protein where the residue presumably could not interact directly with amino acid residues lining the active site gorge. Homologous modelling of housefly AChE, based on the high resolution crystal structure of fruit fly (Drosophila melanogaster) AChE, indicated that D422 may be involved in a salt bridge with H341. The distance between D422 and H341 is predicted to be 3.8 Angstrom capital A, ring, and modification of the charge on the side chain of D422 (D to V) would likely affect the profile of the acyl pocket via the Y339 component. Such a structural change in the acyl pocket in the mutant may underlie the decreased affinity of AChE for CBs.
