Rubidium marking of Anopheles mosquitoes detectable by field-capable X-ray spectrometry

We present a mosquito marking technique suitable for mark-release-recapture that can be used with a hand-held, portable X-ray fluorescence (XRF) spectrometer, which is practical for field measurements. Third instar Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) and Anopheles stephensi Liston larvae were cultured to pupation in water containing rubidium (Rb) Cl at concentrations up to 1000 p.p.m. Rb. Anopheles gambiae larvae survived to adulthood at concentrations as high as 1000 p.p.m. Rb but suffered pupal mortality and reduced adult longevity at high concentrations. We were able to culture An. stephensi at Rb concentrations as high as 300 p.p.m. The presence of Rb in adults was evaluated using a portable XRF analyser, and we were able to reliably detect Rb above background levels in 10-day-old females and 4-day-old males at concentrations causing minimal pupal or adult mortality. We observed that Rb marking was not permanent, and the concentration declined significantly as adults aged. The low cost of labelling with RbCl and the field portability of the spectrometer provide a useful means for labelling mosquitoes via breeding sites or in the laboratory for mark-release-recapture experiments.