The wings of the ‘low-crypsis’ targets were uniformly printed with the lighter colour. The high-crypsis targets were expected to be more cryptic than the low-crypsis selleck chemical targets because they better matched the background, and were also potentially disruptive because of the presence of edge-intersecting patches (Stevens & Cuthill, 2006). The pastry in both the high and low-crypsis targets was dyed with 1 mL of black Wilton® gel icing colour
(http://www.wilton.com/) per 500 g pastry. The wings of the white palatable controls had no colour pattern printed on them, and the pastry (white in colour) was not dyed. The remaining two prey types were modified to have either a low (0.6 g quinine hydrochloride, 1.2 g ground mustardseed, BTK activity 0.012 g Bitrex per 500 g pastry) or high (1.5 g quinine hydrochloride, 3 g ground mustardseed, 0.3 g Bitrex per 500 g pastry) level of unpalatability. Quinine hydrochloride has been shown to be aversive to wild avian predators when combined with pastry (Speed et al., 2000), and is chemically similar to quinine compounds found in species of aposematic insects, arachnids and other arthropods (Eisner, Eisner & Siegler, 2005). Quinine
compounds are not toxic to birds, but are bitter tasting and elicit an emetic response at high doses (Alcock, 1970). Bitrex is a bitter-tasting chemical that has been shown to elicit an aversive response in birds (Skelhorn & Rowe, 2009, 2010), but
is not toxic or emetic even at very large doses (Schafer, Bowles & Hurlbut, 1983), so its only role was to provide an unpleasant or aversive taste to predators. The low and high unpalatability treatments were given conspicuous wings coloured either red or yellow depending on the site, to control for possible pre-existing predator colour find more preferences. In sites 1 and 2, the prey with a low level of unpalatability were given yellow wings while highly unpalatable prey were given red wings; these colours were reversed in sites 3 and 4. Both types of unpalatable pastry were dyed with 1 mL of orange Wilton® gel icing colour per 500 g pastry. Trials were conducted for 5 weeks. Each week, one transect was laid in each of the four sites. Each transect contained 12 replicates of the five prey types, for a total of 60 prey items per transect, or 240 per week over all four sites. Individual prey targets were stapled to tree trunks at a height of 2 m, with the paper wings covering the pastry bodies. Only deciduous trees with a diameter greater than 10 cm were used, and trees with prey targets were a minimum of 3 m apart. Transects were left out for 4 days, and prey targets were surveyed at 24, 48, 72 and 96 h for signs of predation by avian predators.