Topical Testing Assay: A Technique to Evaluate the Insecticidal Potential of Dopamine Receptor Antagonists on Adult Female Mosquitoes by Topical Application

Published: April 30, 2023

Abstract

Source: Brito-Sierra, C.A. et al., Protocols for Testing the Toxicity of Novel Insecticidal Chemistries to Mosquitoes. J. Vis. Exp. (2019).

In this video, we demonstrate the topical testing assay to determine the toxicity of a potential insecticide on adult female mosquitoes. Using a dopamine receptor antagonist that causes lethal inhibition of neurotransmission in mosquitoes, this method describes a way to evaluate the toxic effects of insecticides upon topical application.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board 1. Culture of Mosquito Adults NOTE: Insecticide susceptible strains of mosquitoes are available from the Malaria Research and Reagent Reference Repository. Recommended strains are as follows: Aedes aegypti Liverpool (LVP) strain, Anopheles gambiae Kisumu (KISUMU1) strain, and…

Representative Results

Figure 1: Culture of adult mosquitoes. Image showing 20 L plastic bucket used for culture of adult mosquitoes and use of an aspirator to remove mosquitoes. Figure 2. Adult topical assay performed using 4-5-day-old femal…

開示

The authors have nothing to disclose.

Materials

Insecticide susceptible mosquito strains Malaria Research and Reagent Reference Repository https://www.beiresources.org/MR4Home.aspx; Recommended strains: Aedes aegypti Liverpool (LVP) strain, Anopheles gambiae Kisumu (KISUMU1) strain and Culex quinquefasciatus Johannesburg (JHB) strain
Acetone Mallinckrodt Chemical CAS 67-64-1 Use for dilutions and control
Amitriptyline Sigma-Aldrich CAS: 549-18-8 Can be diluted in acetone
Bifenthrin Sigma-Aldrich CAS: 51529-01-2 Synthetic pyrethorid used as positive control
Micro-applicator Burkard Manufacturing Co. Tool needed for topical application experiments
24-well cell culture plate with lid Corning Incorporated 3526
Advantage rubber bands Alliance Rubber Co. Used to seal the paper cups
Glass syringe (1 ml) Becton Dickinson and Co. 512004 Needed for the micro-applications. Glass is better than plastic
Disposable scintillation vials (20 ml) Fisher Scientific 74505-20 Glass vials prevent evaporation
Tulle fabric, white Walmart
Paper cups Dixie Consumer Products LLC. PF15675/13D Used to keep adult mosquitoes in adult topical assays
Petri dishes (150 mm) Corning Life Sciences Used to maintain the mosquitoes "slept" on cold without direct contact with ice
Transfer pipettes Fisher Scientific 13-711-7M Used to sort larvae
Stereo microscope Olympus SZ6045 Used to score larval assays and perform micro-applications
Tweezers (fine) Fontax Used to handle adult mosquitoes

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記事を引用
Topical Testing Assay: A Technique to Evaluate the Insecticidal Potential of Dopamine Receptor Antagonists on Adult Female Mosquitoes by Topical Application. J. Vis. Exp. (Pending Publication), e21170, doi: (2023).

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