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Area of Science:

  • Virology
  • Arthropod-borne viruses
  • Molecular biology

Background:

  • Entebbe bat virus (ENTV) is a bat-associated flavivirus lacking a known arthropod vector.
  • Research on ENTV biology and potential vector transmission is limited by a lack of molecular tools.
  • The complete 3' untranslated region of ENTV was sequenced, and an infectious clone was developed.

Purpose of the Study:

  • To develop molecular tools for studying Entebbe bat virus (ENTV).
  • To investigate the potential for arthropod vector-borne transmission of ENTV.
  • To assess ENTV replication in mosquito cell lines and whole mosquitoes.

Main Methods:

  • Sequencing of the complete 3' untranslated region of ENTV.
  • Development of an infectious clone of ENTV.
  • Replication studies in vertebrate and mosquito cell lines (Aedes aegypti, Aedes albopictus, Culex tarsalis).
  • Oral exposure and intrathoracic inoculation of Aedes mosquitoes with ENTV.
  • Phylogenetic and bioinformatics analyses.

Main Results:

  • ENTV replicated similarly to the parental isolate in vertebrate cells.
  • High-titer replication of ENTV was observed in Aedes aegypti and Aedes albopictus cell lines, but not in Culex tarsalis cells.
  • Phylogenetic analysis suggested a potential association between ENTV and mosquitoes.
  • Oral exposure of Aedes mosquitoes to ENTV resulted in negative infection, despite high replication after intrathoracic inoculation.
  • ENTV replicated in the mosquito body after IT inoculation but did not disseminate into saliva.

Conclusions:

  • Aedes aegypti and Aedes albopictus are unlikely to be competent vectors for ENTV transmission due to strong midgut infection barriers.
  • ENTV exhibits high molecular compatibility with Aedes cell lines but faces significant barriers to transmission.
  • The developed infectious clone and findings facilitate further research into ENTV transmission and maintenance mechanisms.