Barcoded Asaia bacteria enable mosquito in vivo screens and identify novel systemic insecticides and inhibitors of malaria transmission

PLoS Biol. 2021 Dec 20;19(12):e3001426. doi: 10.1371/journal.pbio.3001426. eCollection 2021 Dec.


This work addresses the need for new chemical matter in product development for control of pest insects and vector-borne diseases. We present a barcoding strategy that enables phenotypic screens of blood-feeding insects against small molecules in microtiter plate-based arrays and apply this to discovery of novel systemic insecticides and compounds that block malaria parasite development in the mosquito vector. Encoding of the blood meals was achieved through recombinant DNA-tagged Asaia bacteria that successfully colonised Aedes and Anopheles mosquitoes. An arrayed screen of a collection of pesticides showed that chemical classes of avermectins, phenylpyrazoles, and neonicotinoids were enriched for compounds with systemic adulticide activity against Anopheles. Using a luminescent Plasmodium falciparum reporter strain, barcoded screens identified 48 drug-like transmission-blocking compounds from a 400-compound antimicrobial library. The approach significantly increases the throughput in phenotypic screening campaigns using adult insects and identifies novel candidate small molecules for disease control.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetobacteraceae / genetics
  • Animals
  • Anopheles / genetics
  • Anopheles / microbiology
  • Antimalarials / pharmacology
  • DNA Barcoding, Taxonomic / methods*
  • Drug Evaluation, Preclinical / methods*
  • Insecticides
  • Malaria / parasitology
  • Malaria / prevention & control*
  • Malaria / transmission
  • Mosquito Vectors / microbiology
  • RNA, Ribosomal, 16S / genetics


  • Antimalarials
  • Insecticides
  • RNA, Ribosomal, 16S

Grants and funding

This work was financially supported by the Bill and Melinda Gates Foundation ( through grants OPP1067662 and OPP1118462 to KJD. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Under the grant conditions of the Foundation, a Creative Commons Attribution 4.0 Generic License has already been assigned to the Author Accepted Manuscript version.