A novel spectral representation based on fractional Fourier transform (FrFT) is proposed and applied to birdcall analysis. The FrFT-based spectrogram of a signal is derived and compared with its FT-based counterpart, and the spectrum gathering method is used to show the energy distribution related to the pitch frequency. The fixed transform order and adaptive orders for FrFT are tested. The fixed order can be obtained empirically or calculated according to the known chirp rate. The adaptive optimal orders are determined by using ambiguity function. Experimental results with birdcalls show that the FrFT-based spectrogram with an optimal transform order has higher resolution than its STFT-based counterpart, and the better performance can be achieved if adaptive orders are used.