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Background  Prior to a major release campaign of sterile insects, including the sterile insect technique, male mosquitoes must be marked and released (small scale) to determine key parameters including wild population abundance, dispersal and survival. Marking insects has been routinely carried out for over 100 years; however, there is no gold standard regarding the marking of specific disease-transmitting mosquitoes including  Anopheles arabiensis ,  Aedes aegypti  and  Aedes albopictus . The research presented offers a novel dusting technique and optimal dust colour and quantities, suitable for small-scale releases, such as mark-release-recapture studies.    Methods  We sought to establish a suitable dust colour and quantity for batches of 100 male  An. arabiensis , that was visible both by eye and under UV light, long-lasting and did not negatively impact longevity. A set of lower dust weights were selected to conduct longevity experiments with both  Ae. aegypti  and  Ae. albopictus  to underpin the optimal dust weight. A further study assessed the potential of marked male  An. arabiensis  to transfer their mark to undusted males and females.    Results  The longevity of male  An. arabiensis  marked with various dust colours was not significantly reduced when compared to unmarked controls. Furthermore, the chosen dust quantity (5 mg) did not negatively impact longevity ( P  = 0.717) and provided a long-lasting mark. Dust transfer was found to occur from marked  An. arabiensis  males to unmarked males and females when left in close proximity. However, this was only noticeable when examining individuals under a stereomicroscope and thus deemed negligible. Overall, male  Ae. aegypti  and  Ae. albopictus  displayed a greater sensitivity to dusting. Only the lowest dust weight (0.5 mg) did not significantly reduce longevity ( P  = 0.888) in  Ae. aegypti , whilst the lowest two dust weights (0.5 and 0.75 mg) had no significant impact on longevity ( P  = 0.951 and 0.166, respectively) in  Ae. albopictus .    Conclusion  We have devised a fast, inexpensive and simple marking method and provided recommended dust quantities for several major species of disease-causing mosquitoes. The novel technique provides an evenly distributed, long-lasting mark which is non-detrimental. Our results will be useful for future MRR studies, prior to a major release campaign.

A standardised method of marking male mosquitoes with fluorescent dust