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Microbiology JoVE (Open Access) Citable · DOI

Forced Salivation As a Method to Analyze Vector Competence of Mosquitoes

DOI: 10.3791/57980-v
What you'll learn
  • ✓ Perform forced salivation assay to detect viral transmission in mosquitoes
  • ✓ Prepare infected mosquitoes via artificial blood meal containing Zika virus
  • ✓ Process and analyze saliva samples for vector competence quantification
  • ✓ Compare transmission potential across Aedes and Culex species
Protocol

For efficient control of mosquito borne virus transmission, the knowledge of the vector potential of respective mosquitoes is of particular interest. We describe forced salivation as a method to analyze vector competence of Aedes albopictus and three different Culex taxa for the transmission of Zika virus.

Difficulty
advanced
Total time
~3 weeks (14–21 days post-infection per cohort)
Model organism
Aedes albopictus, Culex pipiens, Culex modestus, Culex torrentium (mosquitoes)
Biosafety
BSL-2

Steps

1
Provide infected blood meal to mosquitoes

Feed mosquitoes an artificial blood meal containing Zika virus to establish infection. Allow sufficient time for viral replication.

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2
Conduct forced salivation assay

On day 14 or 21 post-infection, perform forced salivation to extract saliva from infected mosquitoes. This measures viral shedding in salivary glands.

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3
Process collected saliva samples

Collect, prepare, and process saliva samples for downstream analysis, including RNA extraction or viral quantification.

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4
Analyze vector competence by species

Compare intrinsic incubation time (IIT) and viral RNA concentration across Aedes and Culex taxa to assess differential transmission potential.

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