Abstract

BackgroundControl of mosquitoes requires the ability to evaluate new insecticides and to monitor resistance to existing insecticides. Monitoring tools should be flexible and low cost so that they can be deployed in remote, resource poor areas. Ideally, a bioassay should be able to simulate transient contact between mosquitoes and insecticides, and it should allow for excito-repellency and avoidance behaviour in mosquitoes. Presented here is a new bioassay, which has been designed to meet these criteria. This bioassay was developed as part of the Mosquito Contamination Device (MCD) project and, therefore, is referred to as the MCD bottle bioassay.MethodsPresented here are two experiments that serve as a proof-of-concept for the MCD bottle bioassay. The experiments used four insecticide products, ranging from fast-acting, permethrin-treated, long-lasting insecticide nets (LLINs) that are already widely used for malaria vector control, to the slower acting entomopathogenic fungus, Beauveria bassiana, that is currently being evaluated as a prospective biological insecticide. The first experiment used the MCD bottle to test the effect of four different insecticides on Anopheles stephensi with a range of exposure times (1 minute, 3 minutes, 1 hour). The second experiment is a direct comparison of the MCD bottle and World Health Organization (WHO) cone bioassay that tests a subset of the insecticides (a piece of LLIN and a piece of netting coated with B. bassiana spores) and a further reduced exposure time (5 seconds) against both An. stephensi and Anopheles gambiae. Immediate knockdown and mortality after 24 hours were assessed using logistic regression and daily survival was assessed using Cox proportional hazards models.ResultsAcross both experiments, fungus performed much more consistently than the chemical insecticides but measuring the effect of fungus required monitoring of mosquito mortality over several days to a week. Qualitatively, the MCD bottle and WHO cone performed comparably, although knockdown and 24 hour mortality tended to be higher in some, but not all, groups of mosquitoes exposed using the WHO cone.ConclusionThe MCD bottle is feasible as a flexible, low-cost method for testing insecticidal materials. It is promising as a tool for testing transient contact and for capturing the effects of mosquito behavioural responses to insecticides.

Highlights

  • Control of mosquitoes requires the ability to evaluate new insecticides and to monitor resistance to existing insecticides

  • The two methods differ in that the World Health Organization (WHO) tube relies on equipment that must be acquired from a single source, while the Centers for Disease Control (CDC) assay uses glass bottles that are readily available as laboratory equipment and can be prepared on site by the end user

  • The WHO wireball assay and tunnel test can be used to evaluate insecticide treated bed nets (ITNs), but the two methods differ in an important way: the wireball assay is a contact bioassay like the cone test, where mosquitoes are held in contact with the material, while the tunnel test incorporates a behavioural component by releasing a mosquito in a tunnel with a live host behind a piece of experimentally damaged netting

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Summary

Introduction

Control of mosquitoes requires the ability to evaluate new insecticides and to monitor resistance to existing insecticides. The standard tools to test insecticides, insecticide treated materials, and monitor resistance are the World Health Organization (WHO) tube, the WHO cone, the Centers for Disease Control (CDC) bottle assay [2] and, to a lesser extent, the WHO wireball assay and tunnel test [3,4,5]. These methods are used for different kinds of tests, and all of them have a different set of benefits and drawbacks. It is worth noting that all of these tools use a protocol that calls for a relatively long exposure time (depending on the test, 3 minutes to 1 hour exposures or up to 12–15 hours in the case of the tunnel test) and relatively quick read outs – knockdown measured 1 hour post exposure and mortality measured 24 hours post exposure

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