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Table 3 Guidelines on repellent testing adapted from [78]

From: Plant-based insect repellents: a review of their efficacy, development and testing

WHOPES approved repellent testing methodology

Laboratory Testing

Use 20% deet in ethanol as a positive comparison

Human subjects preferable to reflect the end user

Before the test the test area of skin should be washed with unscented soap then rinsed with 70% ethanol / isopropyl alcohol

Mosquitoes should be reared under standard 27 ± 2 C temperature, ≥80 ± 10% relative humidity, and a 12:12 (light:dark) photoperiod.

Mosquitoes should be 3 to 5 days old, nulliparous females, starved for 12 hours preceding the test

Tests should be conducted with three or more species

40 x 40 x 40 cm cages with 50 – 100 mosquitoes for effective dose testing

40 x 40 x 40 cm cages with 200 - 250 mosquitoes for complete protection time testing

Control arms should be used to estimate mosquito readiness to feed

Treatment arms should be offered to mosquitoes after avidity has been measured

Field Testing

Use 20% deet in ethanol as a positive comparison

Human subjects preferable to reflect the end user

Before the test the test area of skin should be washed with unscented soap then rinsed with 70% ethanol / isopropyl alcohol

Volunteers should sit >20 metres apart

Design should be completely randomised

Trials should be conducted with medium biting pressures of representative vector species

All participants should be recruited on informed consent from the local area and be provided with malaria prophylaxis

In all testing monitoring of adverse effects should be carried out