Abstract

BackgroundThe World Health Organization (WHO) cone bioassay plays an integral role in the evaluation of the efficacy of long-lasting insecticidal nets as well as insecticides used in indoor residual spraying. The test is used on a variety of treated substrates, such as pieces of bed nets, mud, cement and wood. The cone setup assumes a wide variety of angles under different settings in which it is applied. However, the guidelines provided for the performance of the assay do not specify the angle at which the test must be performed.MethodsLaboratory colonies of Anopheles gambiae Kisumu-1 and Anopheles stephensi STI were tested in the WHO cone bioassay at four different angles (0°, 45°, 60° and 90°) following the WHO guidelines against net pieces of Olyset Plus and Netprotect. The tests were repeated after 20 washes of the nets. Individual mosquitoes were also exposed at 0° and 60° and the amount of time each spent in contact with the net was recorded.ResultsMosquitoes spent more time on the net at 60° as compared to 0° (coefficient = 45.8, 95 % CI 34.6–55.6, p < 0.001) and were more likely to die when the test was done at 45° (OR 3.3, 95 % CI 1.7–6.3, p = 0.001), 60° (OR 3.1, 95 % CI 1.7–5.9, p < 0.001) and 90° (OR 6.0, 95 % CI 1.9–18.5, p = 0.002) as compared to 0°.ConclusionThe angle at which the test is performed significantly affects the amount of time mosquitoes spend resting on the nets, and subsequently mortality. Angle must thus be considered as an important component in the performance of the assay and duly incorporated into the guidelines.Electronic supplementary materialThe online version of this article (doi:10.1186/s12936-016-1303-9) contains supplementary material, which is available to authorized users.

Highlights

  • The World Health Organization (WHO) cone bioassay plays an integral role in the evaluation of the efficacy of long-lasting insecticidal nets as well as insecticides used in indoor residual spraying

  • Insecticide-treated bed nets (ITNs) have been shown to effectively reduce malaria morbidity and mortality [1] and the last years have seen many intervention programs being put in place to distribute and promote the use of long-lasting insecticidal nets (LNs) in malaria endemic countries [2,3,4,5]

  • The results from the current study provide evidence that the angle at which the WHO cone bioassay is performed considerably affects the time mosquitoes spend in contact with the net, and subsequently 24 h mortality

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Summary

Introduction

The World Health Organization (WHO) cone bioassay plays an integral role in the evaluation of the efficacy of long-lasting insecticidal nets as well as insecticides used in indoor residual spraying. WHO instructs that nets would first have to meet the criteria of WHO cone bioassay before they are passed to go through phase II testing [7] It is the recommended assay for testing the efficacy and irritant or excito-repellent properties of insecticide-treated substrates. It plays a pivotal role in IRS as it is used to test formulations of insecticides on various substrates such as mud, cement, plywood and other materials commonly used for building [8]. In 2013, the WHO Pesticide Evaluation Scheme (WHOPES) published the latest version of the Guidelines for laboratory and field-testing of long-lasting insecticidal nets [7] which replaced the earlier version published in 2005 [9]

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