Advertisement

Fly Models of Vibrio cholerae Infection and Colonization

  • Alexandra E. PurdyEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1839)

Abstract

Studies of Vibrio cholerae pathogenesis in the context of novel eukaryotic model systems have expanded our understanding of genes that underlie V. cholerae interactions with humans, as well as host organisms in the environment. These model systems have also helped uncover new functions for many gene products, revealing previously unknown virulence mechanisms. The Drosophila model for V. cholerae infection is a powerful tool for discovering new genetic pathways that govern bacterial physiology and colonization in the arthropod gastrointestinal tract. Assays to measure both virulence and colonization have been established and are easily adopted in labs unfamiliar with Drosophila work. Experiments to compare survival of flies colonized with different bacterial mutants are simple to perform and can be completed in less than a week, allowing colonization to be quantified and localized easily. The availability of molecular and genetic tools for the fly enables further exploration of host factors that restrict V. cholerae colonization and invasive infection. Based on the Drosophila system, a house fly (Musca domestica) model of V. cholerae colonization has also been developed. The new house fly model may prove a useful tool for examining V. cholerae infection dynamics in the context of a host carrying a complex microbial community, with a fundamentally different ecology that may increase its chances of acting as a vector for cholera disease.

Key words

Drosophila melanogaster Fruit fly Vibrio cholerae Virulence Colonization Survival Confocal microscopy House fly Musca domestica 

Notes

Acknowledgments

We would like to thank Scott Keating and John Stoffolano (University of Massachusetts Amherst) for generously providing their protocols for maintaining house fly colonies and Ethan Graf for commenting on the manuscript and sharing the fly food protocol. Many thanks to Paula Watnick for her insights and discussions throughout a postdoctoral fellowship in her laboratory that introduced me to the Drosophila model of Vibrio cholerae infection, and for her continued support.

References

  1. 1.
    Bonfini A, Liu X, Buchon N (2016) From pathogens to microbiota: how Drosophila intestinal stem cells react to gut microbes. Dev Comp Immunol 64:22–38.  https://doi.org/10.1016/j.dci.2016.02.008 CrossRefPubMedGoogle Scholar
  2. 2.
    Dobson AJ, Chaston JM, Newell PD et al (2015) Host genetic determinants of microbiota-dependent nutrition revealed by genome-wide analysis of Drosophila melanogaster. Nat Commun 6:6312. https://doi.org/10.1038/ncomms7312
  3. 3.
    Broderick NA, Buchon N, Lemaitre B (2014) Microbiota-induced changes in Drosophila melanogaster Host gene expression and gut morphology. MBio 5:e01117.  https://doi.org/10.1128/mBio.01117-14 CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Graczyk TK, Knight R, Gilman RH, Cranfield MR (2001) The role of non-biting flies in the epidemiology of human infectious diseases. Microbes Infect 3:231–235CrossRefPubMedGoogle Scholar
  5. 5.
    Greenberg B (1973) Flies and disease. Princeton University Press, Princeton, NJGoogle Scholar
  6. 6.
    Blow NS, Salomon RN, Garrity K et al (2005) Vibrio cholerae infection of Drosophila melanogaster mimics the human disease cholera. PLoS Pathog 1:e8.  https://doi.org/10.1371/journal.ppat.0010008 CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Berkey CD, Blow N, Watnick PI (2009) Genetic analysis of Drosophila melanogaster susceptibility to intestinal Vibrio cholerae infection. Cell Microbiol 11:461–474.  https://doi.org/10.1111/j.1462-5822.2008.01267.x CrossRefPubMedGoogle Scholar
  8. 8.
    Guichard A, Cruz-Moreno B, Aguilar B et al (2013) Cholera toxin disrupts barrier function by inhibiting exocyst-mediated trafficking of host proteins to intestinal cell junctions. Cell Host Microbe 14:294–305.  https://doi.org/10.1016/j.chom.2013.08.001 CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Wang Z, Hang S, Purdy AE, Watnick PI (2013) Mutations in the IMD pathway and mustard counter Vibrio cholerae suppression of intestinal stem cell division in Drosophila. MBio 4:e00337–e00313.  https://doi.org/10.1128/mBio.00337-13 CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Purdy AE, Watnick PI (2011) Spatially selective colonization of the arthropod intestine through activation of Vibrio cholerae biofilm formation. Proc Natl Acad Sci 108:19,737–19,742.  https://doi.org/10.1073/pnas.1111530108 CrossRefGoogle Scholar
  11. 11.
    Hang S, Purdy AE, Robins WP et al (2014) The acetate switch of an intestinal pathogen disrupts host insulin signaling and lipid metabolism. Cell Host Microbe 16:592–604.  https://doi.org/10.1016/j.chom.2014.10.006 CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Soler-Bistué A, Mondotte JA, Bland MJ et al (2015) Genomic location of the major ribosomal protein gene locus determines Vibrio cholerae global growth and infectivity. PLoS Genet 11:e1005156.  https://doi.org/10.1371/journal.pgen.1005156 CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    El Bassiony GML, Luizzi V, Nguyen D et al (2016) Laboratory infection of the adult house fly (Musca domestica) by Vibrio cholerae. Med Vet Entomol 30:392–402CrossRefPubMedGoogle Scholar
  14. 14.
    Fotedar R (2001) Vector potential of houseflies (Musca domestica) in the transmission of Vibrio cholerae in India. Acta Trop 78:31–34CrossRefPubMedGoogle Scholar
  15. 15.
    Gill CA, Lal RB (1931) The epidemiology of cholera, with special reference to transmission. A preliminary report. Indian J Med Res 18:1255–1297Google Scholar
  16. 16.
    Oo KN, Sebastian AA, AYE T (1989) Carriage of enteric bacterial pathogens by house flies in Yangon, Myanmar. J Diarrhoeal Dis Res 7:81–84Google Scholar
  17. 17.
    Scott JG, Warren WC, Beukeboom LW et al (2014) Genome of the house fly, Musca domestica L., a global vector of diseases with adaptations to a septic environment. Genome Biol 15:466.  https://doi.org/10.1186/s13059-014-0466-3 CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    (2003) Arthropod containment levels (ACLs). Vector-Borne Zoonotic Dis 3:75–90.  https://doi.org/10.1089/153036603322163475
  19. 19.
    (2003) Risk assessment for arthropod vectors. Vector-Borne Zoonotic Dis 3:69–73.  https://doi.org/10.1089/153036603322163466
  20. 20.
    Greenberg B (1971) Flies and disease. Princeton University Press, Princeton, NJGoogle Scholar
  21. 21.
    Bansal R, Hulbert S, Schemerhorn B, Reese JC, Whitworth RJ, Stuart JJ, Chen M-S, Ho PL (2011) Hessian fly-associated bacteria: transmission, essentiality, and composition. PLoS ONE 6(8):e23170CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of BiologyAmherst CollegeAmherstUSA

Personalised recommendations