A non-destructive sugar-feeding assay for parasite detection and estimating the extrinsic incubation period of Plasmodium falciparum in individual mosquito vectors

Edwige Guissou*, Jessica L. Waite, Matthew Jones, Andrew S. Bell, Eunho Suh, Koudraogo B. Yameogo, Nicaise Djègbè, Dari F. Da, Domonbabele F.D.S. Hien, Rakiswende S. Yerbanga, Anicet G. Ouedraogo, Kounbobr Roch Dabiré, Anna Cohuet, Matthew B. Thomas, Thierry Lefèvre

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review


Despite its epidemiological importance, the time Plasmodium parasites take to achieve development in the vector mosquito (the extrinsic incubation period, EIP) remains poorly characterized. A novel non-destructive assay designed to estimate EIP in single mosquitoes, and more broadly to study Plasmodium–Anopheles vectors interactions, is presented. The assay uses small pieces of cotton wool soaked in sugar solution to collect malaria sporozoites from individual mosquitoes during sugar feeding to monitor infection status over time. This technique has been tested across four natural malaria mosquito species of Africa and Asia, infected with Plasmodium falciparum (six field isolates from gametocyte-infected patients in Burkina Faso and the NF54 strain) and across a range of temperatures relevant to malaria transmission in field conditions. Monitoring individual infectious mosquitoes was feasible. The estimated median EIP of P. falciparum at 27 °C was 11 to 14 days depending on mosquito species and parasite isolate. Long-term individual tracking revealed that sporozoites transfer onto cotton wool can occur at least until day 40 post-infection. Short individual EIP were associated with short mosquito lifespan. Correlations between mosquito/parasite traits often reveal trade-offs and constraints and have important implications for understanding the evolution of parasite transmission strategies.

Original languageEnglish
Article number9344
JournalScientific Reports
Issue number9394
Publication statusPublished - 29 Apr 2021

Bibliographical note

Funding Information:
Thanks to Deonna Soergel for help with dissections, Janet Teeple and Natalie Gahm for mosquito colony rearing assistance, and the Thomas and Read groups, Johanna Ohm, and Elizabeth McGraw for helpful discussion. Thanks to Fhallon Ware-Gilmore for sharing some infected mosquitoes. We thank gametocyte carrier volunteers for participating in this study and the local authorities for their support. Thanks to the infectivity group at the IRSS Bobo Dioulasso for technical support and discussions. MBT, JLW and ES were part-funded by NIH NIAID grant # R01AI110793 and National Science Foundation Ecology and Evolution of Infectious Diseases grant (DEB-1518681). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Publisher Copyright:
© 2021, The Author(s).

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