A better understanding of the oviposition behaviour of malaria vectors might facilitate the development of new vector control tools. However, the factors that guide the aquatic habitat selection of gravid females are poorly understood. The present study explored the relative attractiveness of similar artificial ponds (0.8 m ) aged at varying lengths prior to opening in such a way that wild Anopheles arabiensis could choose between ponds that were freshly set up, or were aged 4 or 17 days old, to lay eggs. Physicochemical parameters, bacterial profile and volatile organic compounds emitted from ponds were investigated over three experimental rounds. Fresh ponds contained on average twice as many An. arabiensis instar larvae (mean 50, 95% confidence interval (CI) = 29-85) as the ponds that had aged 4 days (mean = 24, 95% CI = 14-42) and 17 days (mean = 20, 95% CI: 12-34). Fresh ponds were associated with a significantly higher turbidity combined with higher water temperature, higher nitrite levels and a lower pH and chlorophyll level than the older ponds. Round by round analyses suggested that bacteria communities differed between age groups and also that 4-heptanone, 2-ethylhexanal and an isomer of octenal were exclusively detected from the fresh ponds. These characteristics may be useful with respect to developing attract and kill strategies for malaria vector control.

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The male butterfly Pieris napi produces the anti-aphrodisiac pheromone methyl salicylate (MeS) and transfers it to the female during mating. After mating she releases MeS, when courted by conspecific males, which decreases her attractiveness and the duration of male harassment, thus increasing her time available for egg-laying. In previous studies we have shown that males produced MeS from the amino acid L-phenylalanine (L-Phe) acquired during larval stage. In this study we show that adult males of P. napi can utilize L-Phe and aromatic flower volatiles as building blocks for production of anti-aphrodisiac pheromone and transfer it to females during mating. We demonstrate this by feeding butterflies with stable isotope labelled molecules mixed in sugar solutions, and, to mimic the natural conditions, we fed male butterflies with floral nectar of Bunias orientalis plants treated with labelled L-Phe. The volatiles from butterflies and plants were collected and identified by solid phase micro extraction, gas chromatography and mass spectrometry techniques. Since P. napi is polygamous, males would gain from restoring the titre of MeS after mating and the use of aromatic precursors for production of MeS could be considered as an advantageous trait which could enable butterflies to relocate L-Phe for other needs.