A Survey Of Mutations In The Cochliomyia Hominivorax (diptera: Calliphoridae) Esterase E3 Gene Associated With Organophosphate Resistance And The Molecular Identification Of Mutant Alleles

Cochliomyia hominivorax (Calliphoridae) is one of the most important myiasis-causing flies and is responsible for severe economic losses to the livestock industry throughout the Neotropical region. In Brazil, C. hominivorax has been controlled mainly with organophosphate (OP) insecticides, although the inappropriate use of these chemicals can result in the selection of resistant flies. Changes in carboxylesterase activity have been associated with OP insecticides in some arthopodan species. In this work, we isolated and characterized part of the E3 gene in C. hominivorax (ChαE7), which contained the same substitutions responsible for the acquisition of OP hydrolase activity in Lucilia cuprina (Calliphoridae). Digestion of the polymerase chain reaction products with a restriction enzyme that specifically recognized the mutation site unambiguously differentiated wild and mutated esterase alleles. The PCR-RFLP assay therefore provided a fast, reliable DNA-based method for identifying C. hominivorax individuals with a mutation in the esterase gene. Further bioassays to determine the association of this mutation with OP resistance in C. hominivorax should allow the development of more effective strategies for managing this species. © 2006 Elsevier B.V. All rights reserved.14003/04/15344351Altschul, S.F., Maden, T.L., Schäffer, A.A., Zhang, Z., Miller, W., Lipman, D.J., Gapped BLAST and PSI-BLAST: a new generation of protein data base search programs (1997) Nucleic Acids Res., 25, pp. 3389-3402Bigley, W.S., Plapp Jr., F.W., Esterase activity and susceptibility to parathion at different stages in the life cycle of organophosphorus-resistant and susceptible house flies (1961) J. Econ. Entomol., 54, p. 904Campbell, P.M., Newcomb, R.D., Russel, R.J., Oakeshott, J.G., Two different amino acid substitutions in the ali-esterase E3 confer alternative types of organophosphorus insecticide resistance in the sheep blowfly, Lucilia cuprina (1998) Insect Biochem. Mol. Biol., 28, pp. 139-150Campbell, P.M., Trott, J.T., Claudianos, C., Smyth, K.A., Russel, R.J., Oakeshott, J.G., Biochemistry of esterases associated with organophosphate resistance with comparisons to putative orthologues in other Diptera (1997) Biochem. Genet., 35, pp. 17-40Cassanelli, S., Cerchiari, B., Giannini, S., Bizarro, D., Mazzoni, E., Manicardi, G.C., Use of the RFLP-PCR diagnostic test for characterizing MACE and kdr insecticide resistance in the peach potato aphid Myzus persicae (2005) Pest Manage. Sci., 61, pp. 91-96Claudianos, C., Russell, R.J., Oakeshott, J.G., The same amino acid substitution in orthologous esterases confers organophosphate resistance on the house fly and a blowfly (1999) Insect Biochem. Mol. Biol., 29, pp. 675-686Cygler, M., Schrag, J.D., Sussman, J.L., Harel, M., Silman, I., Gentry, M.K., Doctor, B.P., Relationship between sequence conservation and three-dimensional structure in a large family of esterases, lipases, and related proteins (1993) Protein Sci., 2, pp. 366-382Hall, T.A., BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT (1999) Nucleic Acids Symp. Ser., 41, pp. 95-98Hall, M., Wall, R., Myiasis of human and domestic animals (1995) Adv. Parasitol., 35, pp. 257-334Heidari, R., Devonshire, A.L., Campbell, B.E., Bell, K.L., Dorrian, S.J., Oakeshott, J.G., Russel, R.J., Hydrolysis of pyrethroids by carboxylesterases from Lucilia cuprina and Drosophila melanogaster with active sites modified by in vitro mutagenesis (2005) Insect Biochem. Mol. Biol., 35, pp. 597-609Hemingway, J., The molecular basis of two contrasting metabolic mechanisms of insecticide resistance (2000) Insect Biochem. Mol. Biol., 30, pp. 1009-1015Hemingway, J., Karunaratne, S.H.P.P., Mosquito carboxylesterases: a review of the molecular biology and biochemistry of a major insecticide resistance mechanism (1998) Med. Vet. Entomol., 12, pp. 1-12Hughes, P.B., Mckenzie, J.A., Insecticide resistance in the Australian sheep blowfly, Lucilia cuprina: speculation, science and strategies (1987) Combating Resistance to Xenobiotics: Biological and Chemical Approaches, pp. 162-177. , Ford M.G., Holloman D.W., Khambay B.P.S., and Sawicki R.M. (Eds), Ellis Horwood, ChichesterHughes, P.B., Raftos, D.A., Genetics of an esterase associated with resistance to organophosphorus insecticides in the sheep blowfly, Lucilia cuprina (Wiedemann) (Diptera: Calliphoridae) (1985) Bull. Entomol. Res., 75, pp. 535-544Infante, M.E., Azeredo-Espin, A.M.L., Genetic variability in mitochondrial DNA of screwworm, Cochliomyia hominivorax (Diptera: Calliphoridae), from Brazil (1995) Biochem. Genet., 33, pp. 737-756Lockridge, O., Blong, R.M., Masson, P., Froment, M.T., Millard, C.B., Broomfield, C.A., A single amino acid substitution, Gly 117His, confers phosphotriesterase (organophosphorus acid anhydride hydrolase) activity on human butyrylcholinesterase (1997) Biochemistry, 36, pp. 786-795Newcomb, R.D., Campbell, P.M., Ollis, D.L., Cheah, E., Russel, R.J., Oaheshott, J.G., A single amino acid substitution converts a carboxylesterase to an organophosphate hydrolase and confers insecticide resistance on a blowfly (1997) Proc. Natl. Acad. Sci. U.S.A., 94, pp. 7464-7468Newcomb, R.D., Campbell, P.M., Russell, R.J., Oakeshott, J.G., cDNA cloning, baculovirus-expression and kinetic properties of the esterase, E3, involved in organophosphorus insecticide resistance in Lucilia cuprina (1997) Insect Biochem. Mol. Biol., 27, pp. 15-25Newcomb, R.D., Gleeson, D.M., Yong, C.G., Russell, R.J., Oakeshott, J.G., Multiple mutations and gene duplications conferring organophosphorus insecticide resistance have been selected at the Rop-1 locus of the sheep blowfly, Lucilia cuprina (2005) J. Mol. Evol., 60, pp. 207-220Parker, A.G., Campbell, P.M., Spackman, M.E., Russell, R.J., Oakeshott, J.G., Comparison of an esterase associated with organophosphate resistance in Lucilia cuprina with an orthologue not associated with resistance in Drosophila melanogaster (1996) Pestic. Biochem. Physiol., 55, pp. 85-99Price, N.R., Insect resistance to insecticides: mechanisms and diagnosis (1991) Comp. Biochem. Physiol., 100 (3), pp. 319-326Smyth, K.A., Boyce, T.M., Russell, R.J., Oakeshott, J.G., MCE activities and malathion resistances in field populations of the Australian sheep blowfly (Lucilia cuprina) (2000) Heredity, 84, pp. 63-72Spackman, M.E., Oakeshott, J.G., Smyth, K.A., Medveczky, K.M., Russell, R.J., A cluster of esterase genes on chromosome 3R of Drosophila melanogaster includes homologues of esterase genes conferring insecticide resistance in several Diptera (1994) Biochem. Genet., 32, pp. 39-62Sussman, J.S., Harel, M., Frolov, F., Oefner, C., Goldman, A., Toker, L., Silman, I., Atomic structure of acetylcholinesterase from Torpedo californica: a prototypic acetylcholine-binding protein (1991) Science, 253, pp. 872-879Taskin, V., Kence, M., Göçmen, B., Determination of malathion and diazinon resistance by sequencing the MdαE7 gene from Guatemala, Colombia, Manhattan, and Thailand housefly (Musca domestica) strains (2004) Russ. J. Genet., 40, pp. 377-380Thompson, J.D., Gibson, T.J., Plewniak, F., Jeanmougin, F., Higgins, D.G., The CustalX Windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools (1997) Nucleic Acids Res., 24, pp. 4876-4882Townsend, M.G., Busvine, J.R., The mechanism of malathion resistance in blowfly Chrysomya putoria (1969) Entomol. Exp. Appl., 12, pp. 243-267Van Asperen, K., Oppenoorth, F.J., Organophosphate resistance and esterase activity in houseflies (1969) Entomol. Exp. Appl., 2, pp. 48-57(1965) Myiasis in Man and Animals in the Old World, , Zumpt F. (Ed), London, Butterworths 267 p