The Use of Xenosurveillance to Detect Human Bacteria, Parasites, and Viruses in Mosquito Bloodmeals

Joseph R. Fauver Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado;

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Alex Gendernalik Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado;

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James Weger-Lucarelli Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado;

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Nathan D. Grubaugh Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado;
Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California;

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Doug E. Brackney Center for Vector Biology and Zoonotic Diseases, Connecticut Agricultural Experiment Station, New Haven, Connecticut

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Brian D. Foy Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado;

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Gregory D. Ebel Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, Colorado;

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Infectious disease surveillance is hindered by several factors, including limited infrastructure and geographic isolation of many resource-poor regions. In addition, the complexities of sample acquisition, processing, and analysis, even in developed regions, can be rate limiting. Therefore, new strategies to survey human populations for emerging pathogens are necessary. Xenosurveillance is a method that utilizes mosquitoes as sampling devices to search for genetic signatures of pathogens in vertebrates. Previously we demonstrated that xenosurveillance can detect viral RNA in both laboratory and field settings. However, its ability to detect bacteria and parasites remains to be assessed. Accordingly, we fed Anopheles gambiae mosquitoes blood that contained Trypanosoma brucei gambiense and Bacillus anthracis. In addition, we determined whether two additional emerging viruses, Middle East Respiratory Syndrome Coronavirus and Zika virus could be detected by this method. Pathogen-specific real-time reverse transcription polymerase chain reaction was used to evaluate the sensitivity of xenosurveillance across multiple pathogen taxa and over time. We detected RNA from all pathogens at clinically relevant concentrations from mosquitoes processed up to 1 day postbloodfeeding. These results demonstrate that xenosurveillance may be used as a tool to expand surveillance for viral, parasitic, and bacterial pathogens in resource-limited areas.

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Author Notes

Address correspondence to Gregory D. Ebel, Department of Microbiology, Immunology, and Pathology, Colorado State University, 1692 Campus Delivery, Fort Collins, CO 80523. E-mail: gregory.ebel@colostate.edu

Authors’ addresses: Joseph R. Fauver, Alex Gendernalik, James Weger-Lucarelli, Brian D. Foy, and Gregory D. Ebel, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO, E-mails: joseph.fauver@colostate.edu, alghobbes@gmail.com, james.weger@colostate.edu, brian.foy@colostate.edu, and gregory.ebel@colostate.edu. Nathan D. Grubaugh, Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, CA, E-mail: nathan.grubaugh@yahoo.com. Doug E. Brackney, Center for Vector Biology and Zoonotic Diseases, Connecticut Agricultural Experiment Station, New Haven, CT, E-mail: doug.brackney@ct.gov.

Financial support: The projected was supported in part by the CSU Infectious Disease Supercluster Grant “Xenosurveillance: A novel approach for interrogating the human-pathogen landscape in sub-Saharan Africa” awarded to DEB, BDF, and GDE. This project was also supported in part by an Armed Forces Health Surveillance Center grant awarded to the Walter Reed Army Institute (subcontract DEB).

  • 1.

    Breman JG, 2001. The ears of the hippopotamus: manifestations, determinants, and estimates of the malaria burden. Am J Trop Med Hyg 64: 111.

  • 2.

    Crump JA et al., 2013. Etiology of severe non-malaria febrile illness in northern Tanzania: a prospective cohort study. PLoS Negl Trop Dis 7: e2324.

  • 3.

    Jones KE, Patel NG, Levy MA, Storeygard A, Balk D, Gittleman JL, Daszak P, 2008. Global trends in emerging infectious diseases. Nature 451: 990993.

  • 4.

    Grubaugh ND, Sharma S, Krajacich BJ, Fakoli Iii LS, Bolay FK, Diclaro Ii JW, Johnson WE, Ebel GD, Foy BD, Brackney DE, 2015. Xenosurveillance: a novel mosquito-based approach for examining the human-pathogen landscape. PLoS Negl Trop Dis 9: e0003628.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 5.

    Yang Y, Garver LS, Bingham KM, Hang J, Jochim RC, Davidson SA, Richardson JH, Jarman RG, 2015. Feasibility of using the mosquito blood meal for rapid and efficient human and animal virus surveillance and discovery. Am J Trop Med Hyg 93: 13771382.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 6.

    Kelly-Hope LA, Hemingway J, McKenzie FE, 2009. Environmental factors associated with the malaria vectors Anopheles gambiae and Anopheles funestus in Kenya. Malar J 8: 268.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 7.

    Coetzee M, Craig M, le Sueur D, 2000. Distribution of African malaria mosquitoes belonging to the Anopheles gambiae complex. Parasitol Today 16: 7477.

  • 8.

    Pappa V, Reddy M, Overgaard HJ, Abaga S, Caccone A, 2011. Estimation of the human blood index in malaria mosquito vectors in Equatorial Guinea after indoor antivector interventions. Am J Trop Med Hyg 84: 298301.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 9.

    Harbison JE, Mathenge EM, Misiani GO, Mukabana WR, Day JF, 2006. A simple method for sampling indoor-resting malaria mosquitoes Anopheles gambiae and Anopheles funestus (Diptera: Culicidae) in Africa. J Med Entomol 43: 473479.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 10.

    Bayoh MN et al., 2014. Persistently high estimates of late night, indoor exposure to malaria vectors despite high coverage of insecticide treated nets. Parasit Vectors 7: 380.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 11.

    Barbazan P, Thitithanyanont A, Misse D, Dubot A, Bosc P, Luangsri N, Gonzalez JP, Kittayapong P, 2008. Detection of H5N1 avian influenza virus from mosquitoes collected in an infected poultry farm in Thailand. Vector Borne Zoonotic Dis 8: 105109.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 12.

    Ng TF, Willner DL, Lim YW, Schmieder R, Chau B, Nilsson C, Anthony S, Ruan Y, Rohwer F, Breitbart M, 2011. Broad surveys of DNA viral diversity obtained through viral metagenomics of mosquitoes. PLoS One 6: e20579.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 13.

    Brugman VA, Hernández-Triana LM, Prosser SWJ, Weland C, Westcott DG, Fooks AR, Johnson N, 2015. Molecular species identification, host preference and detection of myxoma virus in the Anopheles maculipennis complex (Diptera: Culicidae) in southern England, UK. Parasit Vectors 8: 421.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 14.

    Kading RC, Biggerstaff BJ, Young G, Komar N, 2014. Mosquitoes used to draw blood for arbovirus viremia determinations in small vertebrates. PLoS One 9: e99342.

  • 15.

    Fernández de Marco M, Brugman VA, Hernández-Triana LM, Thorne L, Phipps LP, Nikolova NI, Fooks AR, Johnson N, 2016. Detection of Theileria orientalis in mosquito blood meals in the United Kingdom. Vet Parasitol 229: 3136.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 16.

    Meyers JI, Gray M, Kuklinski W, Johnson LB, Snow CD, Black WC 4th, Partin KM, Foy BD, 2015. Characterization of the target of ivermectin, the glutamate-gated chloride channel, from Anopheles gambiae. J Exp Biol 218: 14781486.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 17.

    Hirumi H, Hirumi K, 1989. Continuous cultivation of Trypanosoma brucei blood stream forms in a medium containing a low concentration of serum protein without feeder cell layers. J Parasitol 75: 985989.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 18.

    Weger-Lucarelli J et al., 2016. Vector competence of American mosquitoes for three strains of Zika virus. PLoS Negl Trop Dis 10: e0005101.

  • 19.

    Jensen BC, Sivam D, Kifer CT, Myler PJ, Parsons M, 2009. Widespread variation in transcript abundance within and across developmental stages of Trypanosoma brucei. BMC Genomics 10: 482.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 20.

    Qi Y, Patra G, Liang X, Williams LE, Rose S, Redkar RJ, DelVecchio VG, 2001. Utilization of the rpoB gene as a specific chromosomal marker for real-time PCR detection of Bacillus anthracis. Appl Environ Microbiol 67: 37203727.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 21.

    Lu X, Whitaker B, Sakthivel SK, Kamili S, Rose LE, Lowe L, Mohareb E, Elassal EM, Al-sanouri T, Haddadin A, Erdman DD, 2014. Real-time reverse transcription-PCR assay panel for Middle East respiratory syndrome coronavirus. J Clin Microbiol 52: 6775.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 22.

    Robert SL, Olga LK, Janeen JL, Jason OV, Amy JL, Alison JJ, Stephanie MS, Mark RD, 2008. Genetic and serologic properties of Zika virus associated with an epidemic, Yap State, Micronesia, 2007. Emerg Infect Dis J 14: 1232.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 23.

    Smith CE, 1957. Diagnostic procedures for virus and rickettsial diseases (2nd ed.). Am J Public Health Nations Health 47: 249250.

  • 24.

    Turell MJ, Knudson GB, 1987. Mechanical transmission of Bacillus anthracis by stable flies (Stomoxys calcitrans) and mosquitoes (Aedes aegypti and Aedes taeniorhynchus). Infect Immun 55: 18591861.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 25.

    Waggoner JJ, Gresh L, Vargas MJ, Ballesteros G, Tellez Y, Soda KJ, Sahoo MK, Nuñez A, Balmaseda A, Harris E, Pinsky BA, 2016. Viremia and clinical presentation in Nicaraguan patients infected with Zika virus, chikungunya virus, and dengue virus. Clin Infect Dis 63: 15841590.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 26.

    Corman VM et al., 2016. Viral shedding and antibody response in 37 patients with Middle East respiratory syndrome coronavirus infection. Clin Infect Dis 62: 477483.

  • 27.

    Ross R, Thomson D, 1910. A case of sleeping sickness showing regular periodical increase of the parasites disclosed. BMJ 1: 15441545.

  • 28.

    Minard G, Mavingui P, Moro CV, 2013. Diversity and function of bacterial microbiota in the mosquito holobiont. Parasit Vectors 6: 146.

  • 29.

    Horn D, 2014. Antigenic variation in African trypanosomes. Mol Biochem Parasitol 195: 123129.

  • 30.

    Downe AER, Goring NL, West AS, 1963. The influence of size and source of blood meals on rate of digestion of vertebrate serum proteins in mosquitoes (Diptera: Culicidae). J Kans Entomol Soc 36: 200206.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 31.

    Irby WS, Apperson CS, 1989. Immunoblot analysis of digestion of human and rodent blood by Aedes aegypti (Diptera: Culicidae). J Med Entomol 26: 284293.

  • 32.

    West AS, Eligh GS, 1952. The rate of digestion of blood in mosquitoes. Precipitin test studies. Can J Zool 30: 267272.

  • 33.

    Quinones ML, Lines JD, Thomson MC, Jawara M, Morris J, Greenwood BM, 1997. Anopheles gambiae gonotrophic cycle duration, biting and exiting behaviour unaffected by permethrin-impregnated bednets in The Gambia. Med Vet Entomol 11: 7178.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 34.

    Martínez-de la Puente J, Ruiz S, Soriguer R, Figuerola J, 2013. Effect of blood meal digestion and DNA extraction protocol on the success of blood meal source determination in the malaria vector Anopheles atroparvus. Malar J 12: 109.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 35.

    Niare S, Berenger J-M, Dieme C, Doumbo O, Raoult D, Parola P, Almeras L, 2016. Identification of blood meal sources in the main African malaria mosquito vector by MALDI-TOF MS. Malar J 15: 87.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 36.

    CDC, 2014. National Notifiable Diseases Surveillance System (NNDSS). Available at: http://wwwn.cdc.gov/nndss/script/nedss.aspx. Accessed February 19, 2015.

    • PubMed
    • Export Citation
  • 37.

    CDC, 2014. BioSense: National Syndromic Surveillance Program. Available at: http://www.cdc.gov/nssp/biosense/index.html. Accessed February 19, 2015.

    • PubMed
    • Export Citation
  • 38.

    Department of Defense, 2016. Global Emerging Infections Surveillance and Response System. Available at: https://health.mil/Military-Health-Topics/Health-Readiness/Armed-Forces-Health-Surveillance-Branch/Global-Emerging-Infections-Surveillance-and-Response. Accessed October 25, 2016.

    • PubMed
    • Export Citation
  • 39.

    Espino J, Wagner M, Tsui F, Wilson T, 2014. RODS: Real-Time Outbreak and Disease Surveillance. Available at: www.rods.pitt.edu/site/component/option,com_frontpage/Itemid,34/. Accessed February 19, 2015.

    • PubMed
    • Export Citation
  • 40.

    Wolfe N, 2014. Global Viral Forecasting Initiative. Available at: http://globalviral.org/. Accessed February 19, 2015.

    • PubMed
    • Export Citation
  • 41.

    Organization WH, 2014. GOARN: Gloval Outbreak Alert and Response Network. Available at: http://www.who.int/csr/outbreaknetwork/en/. Accessed February 19, 2015.

    • PubMed
    • Export Citation
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