Esterase-mediated spinosad resistance in house flies Musca domestica (Diptera: Muscidae)

  • Yi Zhang
  • Mingcheng Guo
  • Zhuo Ma
  • Chunmei You
  • Xiwu GaoEmail author
  • Xueyan ShiEmail author


Although esterase-mediated spinosad resistance has been proposed for several insects, the associated molecular mechanism remains poorly understood. In this study, we investigated the mechanism of esterase-based spinosad resistance in house flies using a susceptible strain (SSS) and a spinosad-resistant, near-isogenic line (N-SRS). Combined with the synergistic effect of DEF on spinosad in the N-SRS strain, decreased ali-esterase activity in the spinosad-resistant strain has implicated the involvement of mutant esterase in spinosad resistance in house flies. Examination of the carboxylesterase gene MdαE7 in the two strains revealed that four non-synonymous mutations (Trp251-Leu, Asp273-Glu, Ala365-Val, and Ile396-Val) may be associated with spinosad resistance in house flies. Single nucleotide polymorphism analysis further indicated a strong relationship between these four mutations and spinosad resistance. Moreover, quantitative real-time PCR revealed a female-linked MdαE7 expression pattern in the N-SRS strain, which may contribute to sex-differential spinosad resistance in house flies.


Musca domestica Spinosad resistance Caboxylesterase MdαE7 



We thank the National Natural Science Foundation of China and the National Key Research and Development Program of China for the financial support of this study.


This study was funded by the National Natural Science Foundation of China (No. 31672045) and the National Key Research and Development Program of China (2018YFD0200408).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants performed by any of the authors.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Supplementary material

10646_2019_2125_MOESM1_ESM.pdf (692 kb)
Supplementary Information


  1. Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 7:248–254CrossRefGoogle Scholar
  2. Claudianos C, Crone E, Coppin C, Russell RJ, Oakeshott JG (2002) A genomics perspective on mutant aliesterases and metabolic resistance to organophosphates. Acs Symp Ser 808:90–101CrossRefGoogle Scholar
  3. Claudianos C, Russell RJ, Oakeshott JG (1999) The same amino acid substitution in orthologous esterases confers organophosphate resistance on the housefly and a blowfly. Insect Biochem Mol Biol 29:657–686CrossRefGoogle Scholar
  4. Devonshire AL, Heidari R, Bell KL, Campbell PM, Campbell BE, Odgers WA et al. (2003) Kinetic efficiency of mutant carboxylesterases implicated in organophosphate insecticide resistance. Pestic Biochem Physiol 76:1–13CrossRefGoogle Scholar
  5. Dong K, Scott JG (1994) Linkage of kdr-type resistance and the para-homologous sodium channel gene in German cockroaches (Blattella germanica). Insect Biochem Mol Biol 24:647–654CrossRefGoogle Scholar
  6. Heidari R, Devonshire AL, Campbell BE, Bell KL, Dorrian SJ, Oakeshott JG et al. (2004) Hydrolysis of organophosphorus insecticides by in vitro modified carboxylesterase E3 from Lucilia cuprina. Insect Biochem Mol Biol 34:353–363CrossRefGoogle Scholar
  7. Herron GA, Gunning RV, Cottage ELA, Borzatta V, Gobbi C (2014) Spinosad resistance, esterase isoenzymes and temporal synergism in Frankliniella occidentalis (Pergande) in Australia. Pestic Biochem Physiol 114:32–37CrossRefGoogle Scholar
  8. Højland DH, Jensen KMV, Kristensen M (2014) Expression of xenobiotic metabolizing cytochrome P450 genes in a spinosad-resistant Musca domestica L. strain. PLoS one 9:e103689CrossRefGoogle Scholar
  9. Kakani EG, Zygouridis NE, Tsoumani KT, Seraphides N, Zalom FG, Mathiopoulos KD (2010) Spinosad resistance development in wild olive fruit fly Bactrocera oleae (Diptera: Tephritidae) populations in California. Pest Manag Sci 66:447–453Google Scholar
  10. Kaufman PE, Scott JG, Rutz DA (2001) Monitoring insecticide resistance in house flies (Diptera: Muscidae) from New York dairies. Pest Manag Sci 57:514–521CrossRefGoogle Scholar
  11. Liu SS, Li ZM, Liu YQ, Feng MG, Tang ZH (2007) Promoting selection of resistance to spinosad in the parasitoid Catesia plutellae by integrating resistance of hosts to the insecticide into the selection process. Biol Control 41:246–255CrossRefGoogle Scholar
  12. Loughner RL, Warnock DF, Cloyd RA (2005) Resistance of greenhouse, laboratory and native populations of western flower thrips to spinosad. Hort Sci 40:146–149CrossRefGoogle Scholar
  13. Markussen MDK, Kristensen M (2012) Spinosad resistance in female Musca domestica L. from a field-derived population. Pest Manag Sci 68:75–82CrossRefGoogle Scholar
  14. Mckenzie JA, Whitten MJ, Adena MA (1982) The effect of genetic background on the fitness of the diazinon resistance genotypes of the Australian sheep blowfly, Lucilia cuprina. Heredity 49:1–9CrossRefGoogle Scholar
  15. Oppenoorth FJ, van Asperen K (1960) Allelic genes in the housefly producing modified enzymes that cause organophosphate resistance. Science 132:298–299CrossRefGoogle Scholar
  16. Pavela R (2008) Insecticidal properties of several essential oils on the house fly (Musca domestica L.). Phytother Res 22:274–278CrossRefGoogle Scholar
  17. Rehan A, Freed S (2014) Selection, mechanism, cross resistance and stability of spinosad resistance in Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae). Crop Prot 56:10–15CrossRefGoogle Scholar
  18. Reyes M, Rocha K, Alarcón L, Siegwart M, Sauphanor B (2012) Metabolic mechanisms involved in the resistance of field populations of Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) to spinosad. Pestic Biochem Physiol 102:45–50CrossRefGoogle Scholar
  19. Sabourault C, Guzov VM, Koener JF, Claudianos C, Plapp FWJ, Feyereisen R (2001) Overproduction of a P450 that metabolizes diazinon is linked to a loss-of-function in the chromosome 2 ali-esterase (MdaE7) gene in resistant house flies. Insect Mol Biol 10:609–618CrossRefGoogle Scholar
  20. Salgado VL (1998) Studies on the mode of action of spinosad: Insect symptoms and physiological correlates. Pestic Biochem Physiol 60:91–102CrossRefGoogle Scholar
  21. Scott JG (1998) Toxicity of spinosad to susceptible and resistant strains of house flies, Musca domestica. Pestic Sci 54:131–133CrossRefGoogle Scholar
  22. Scott JG, Alefantis TG, Kaufman PE, Rutz DA (2000) Insecticide resistance in house flies from caged-layer poultry facilities. Pest Manag Sci 56:147–153CrossRefGoogle Scholar
  23. Scott JG, Georghiou GP (1985) Rapid development of high-level permethrin resistance in a field-collected strain of house fly (Diptera: Muscidae) under laboratory selection. J Econ Entomol 78:316–319CrossRefGoogle Scholar
  24. Shan C, Zhang Y, Ma Z, Gao X (2016) Inheritance of propoxur resistance in a near-isogenic line of Musca domestica (Diptera: Muscidae). J Econ Entomol 109:873–878CrossRefGoogle Scholar
  25. Shi J, Zhang L, Gao X (2011) Characterisation of spinosad resistance in the housefly Musca domestica (Diptera: Muscidae). Pest Manag Sci 67:335–340CrossRefGoogle Scholar
  26. Shono T, Kasai S, Kamiya E, Kono Y, Scott JG (2002) Genetics and mechanisms of permethrin resistance in the YPER strain of house fly. Pestic Biochem Physiol 73:27–36CrossRefGoogle Scholar
  27. Shono T, Scott JG (2003) Spinosad resistance in the housefly, Musca domestica, is due to a recessive factor on autosome I. Pestic Biochem Physiol 75:1–7CrossRefGoogle Scholar
  28. Sparks TC, Dripps JE, Watson GB, Paroonagian D (2012) Resistance and cross-resistance to the spinosyns—a review and analysis. Pestic Biochem Physiol 102:1–10CrossRefGoogle Scholar
  29. Taskin V, Kence M (2004) The genetic basis of malathion resistance in housefly (Musca domestica L.) strains from Turkey. Genetika 40:1475–1482Google Scholar
  30. van Asperen K (1964) Biochemistry and genetics of esterases in houseflies (Musca domestica) with special reference to the development of resistance to organophosphorus compounds. Ent Exp Appl 7:205–214CrossRefGoogle Scholar
  31. Wang D, Qiu X, Ren X, Niu F, Wang K (2009a) Resistance selection and biochemical characterization of spinosad resistance in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae). Pestic Biochem Physiol 95:90–94CrossRefGoogle Scholar
  32. Wang D, Qiu X, Ren X, Zhang W, Wang K (2009b) Effects of spinosad on Helicoverpa armigera (Lepidoptera: Noctuidae) from China: tolerance status, synergism, and enzymatic responses. Pest Manag Sci 65:1040–1046CrossRefGoogle Scholar
  33. Wang W, Mo J, Cheng J, Zhuang P, Tang Z (2006) Selection and characterization of spinosad resistance in Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae). Pestic Biochem Physiol 84:180–187CrossRefGoogle Scholar
  34. Young SJ, Gunning RV, Moores GD (2005) The effect of piperonyl butoxide on pyrethroids-resistance-associated esterases in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae). Pest Manag Sci 61:397–401CrossRefGoogle Scholar
  35. Zhang L, Gao X, Liang P (2007) Beta-cypermethrin resistance associated with high carboxylesterase activities in a strain of house fly, Musca domestica (Diptera: Muscidae). Pestic Biochem Physiol 89:65–72CrossRefGoogle Scholar
  36. Zhang L, Shi J, Shi X, Liang P, Gao J, Gao X (2010) Quantitative and qualitative changes of the carboxylesterase associated with beta-cypermethrin resistance in the housefly, Musca domestica (Diptera: Muscidae). Comp Biochem Physiol B Biochem Mol Biol 156:6–11CrossRefGoogle Scholar
  37. Zhang Y, Li J, Ma Z, Shan C, Gao X (2018) Multiple mutations and overexpression of the MdaE7 carboxylesterase gene associated with male-linked malathion resistance in housefly, Musca domestica (Diptera: Muscidae). Sci Rep 8:224CrossRefGoogle Scholar
  38. Zhang Y, Wang Y, Ma Z, Zhai D, Gao X, Shi X (2019) Cytochrome P450 monooxygenases-mediated sex-differential spinosad resistance in house flies Musca domestica (Diptera: Muscidae). Pestic Biochem Physiol 157:178–185CrossRefGoogle Scholar
  39. Zhao JZ, Li YX, Collins HL, Gusukuma-Minuto L, Mau RFL, Thompson GD et al. (2002) Monitoring and characterization of diamondback moth (Lepidoptera: Plutellidae) resistance to spinosad. J Econ Entomol 95:430–436CrossRefGoogle Scholar

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© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.State Key Lab Breeding Base for Zhejiang Sustainable Plant Pest Control, MOA Key Lab for Pesticide Residue Detection, Institute of Quality and Standard for Agro-productsZhejiang Academy of Agricultural SciencesHangzhouChina
  2. 2.Department of EntomologyChina Agricultural UniversityBeijingChina
  3. 3.Institute for the Control of AgrochemicalsMinistry of Agriculture and Rural AffairsBeijingChina
  4. 4.Dongcheng Center for Diseases Prevention and ControlBeijingChina

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