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Fficient quality (low background noise) for further sound analysis. Each song of black redstarts can be divided into three distinct parts (part A, B and C, see Fig. 1 and e.g. [47]) with a pause of varying duration between part A and B. We measured the duration of parts A, B and C, the total song and 1326631 the duration of pauses between A and B (Fig. 1). We counted the number of LY-2409021 site elements of part A and C (mean of max. 20 songrenditions). We also determined the frequency JW-74 bandwidth and the maximum frequency of part A, B and C using the automatic parameter measurement function (threshold 220 dB) in Avisoft (mean of max. 10 renditions of high-quality songs). Data were analyzed with R version 2.9.1 [55]. Song before, during, and after the STI was analyzed using general linear mixed models with bird identity as a random effect to control for repeated measures. We analyzed whether the dependent variables (number of songs, song duration, duration of part A, B, C and the pause between part A and B, the number of elements in part A and C and maximum frequency and bandwidth of all parts) were influenced by the Flut/Let treatment, the context (testing phase of the STI) and their interaction. In all cases, dependent variables where transformed if assumptions of normality and/or equality of variances were not met. Significance was accepted at a #0.05 (two-tailed).Testosterone Affects Song ModulationResults Song in SpringMales sang significantly fewer songs during the STI than when singing spontaneously before and after the STI (Fig. 2a, Table 1). Song duration significantly changed in placebo-implanted males, with songs during the STI being shorter than before or after the STI. In Flut/Let-males, song duration remained constant before, during and after the STI (Table 1). The shortening of the song in placebo-implanted males was mainly due to a significantly shorter pause between song part A and B (Table 1), because the durations of the three song parts (A, B and C) did not differ significantly before, during and after the STI (Table 1). Both placebo-implanted and Flut/Let-males sang significantly more elements in song parts A and C during and after the STI than before the STI (Table 1, Figs. 3c, d). This element increase resulted from an increase in the number of elements of the trilled phrases of part A or C, respectively (Fig. 1). By definition, part B did not change with respect to this measure because it consisted of one element only (Fig. 1). Flut/Let-males sang part A with a significantly lower maximum frequency during and after the STI than before the STI. In contrast, the maximum frequency of part A did not change before, during and after the STI in placebo-implanted males (Table 1, Fig. 3a). Both treatment groups sang part B with a significantly higher maximum frequency during the STI than before the STI. Furthermore, the maximum frequency of this part tended to remain high after the STI in placebo-implanted males but not in Flut/Let-males (Table 1). Consequently, placebo-implanted males sang part B with a significantly larger frequency bandwidth during and after the STI than before the STI, while frequency bandwidth of part B did not change in Flut/Let-implanted males (Table 1, Fig. 3b). Maximum frequency and the frequency bandwidth of part C did not change in response to the STI or Flut/Let-treatment (Table 1).increased the number of elements sung in parts of their song in response to the STI. However, Flut/Let males decreased the maximum acousti.Fficient quality (low background noise) for further sound analysis. Each song of black redstarts can be divided into three distinct parts (part A, B and C, see Fig. 1 and e.g. [47]) with a pause of varying duration between part A and B. We measured the duration of parts A, B and C, the total song and 1326631 the duration of pauses between A and B (Fig. 1). We counted the number of elements of part A and C (mean of max. 20 songrenditions). We also determined the frequency bandwidth and the maximum frequency of part A, B and C using the automatic parameter measurement function (threshold 220 dB) in Avisoft (mean of max. 10 renditions of high-quality songs). Data were analyzed with R version 2.9.1 [55]. Song before, during, and after the STI was analyzed using general linear mixed models with bird identity as a random effect to control for repeated measures. We analyzed whether the dependent variables (number of songs, song duration, duration of part A, B, C and the pause between part A and B, the number of elements in part A and C and maximum frequency and bandwidth of all parts) were influenced by the Flut/Let treatment, the context (testing phase of the STI) and their interaction. In all cases, dependent variables where transformed if assumptions of normality and/or equality of variances were not met. Significance was accepted at a #0.05 (two-tailed).Testosterone Affects Song ModulationResults Song in SpringMales sang significantly fewer songs during the STI than when singing spontaneously before and after the STI (Fig. 2a, Table 1). Song duration significantly changed in placebo-implanted males, with songs during the STI being shorter than before or after the STI. In Flut/Let-males, song duration remained constant before, during and after the STI (Table 1). The shortening of the song in placebo-implanted males was mainly due to a significantly shorter pause between song part A and B (Table 1), because the durations of the three song parts (A, B and C) did not differ significantly before, during and after the STI (Table 1). Both placebo-implanted and Flut/Let-males sang significantly more elements in song parts A and C during and after the STI than before the STI (Table 1, Figs. 3c, d). This element increase resulted from an increase in the number of elements of the trilled phrases of part A or C, respectively (Fig. 1). By definition, part B did not change with respect to this measure because it consisted of one element only (Fig. 1). Flut/Let-males sang part A with a significantly lower maximum frequency during and after the STI than before the STI. In contrast, the maximum frequency of part A did not change before, during and after the STI in placebo-implanted males (Table 1, Fig. 3a). Both treatment groups sang part B with a significantly higher maximum frequency during the STI than before the STI. Furthermore, the maximum frequency of this part tended to remain high after the STI in placebo-implanted males but not in Flut/Let-males (Table 1). Consequently, placebo-implanted males sang part B with a significantly larger frequency bandwidth during and after the STI than before the STI, while frequency bandwidth of part B did not change in Flut/Let-implanted males (Table 1, Fig. 3b). Maximum frequency and the frequency bandwidth of part C did not change in response to the STI or Flut/Let-treatment (Table 1).increased the number of elements sung in parts of their song in response to the STI. However, Flut/Let males decreased the maximum acousti.

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