In this work, echo-planar spectroscopic imaging (EPSI) with flyback readout gradient-echo train was implemented in a preclinical MR scanner. The aim of this study is to visualize and quantify the ghost spectral lines produced by two, three and four interleaved echo trains with different amplitudes of the readout gradients, and to investigate the feasibility of the flyback data acquisition in micro-imaging of small animals. Applied multi-slice EPSI sequence utilizes asymmetric gradient-echo train that combines the shortest possible rewind gradients with readout gradients. It simplifies data processing because all echoes are acquired with the same polarity of the readout gradient. The approach with four interleaved gradient-echo trains and with four echoes in each train provides broad spectral bandwidth in combination with narrow receiver bandwidth and a good water-fat signal separation. It improves signal-to-noise ratio without the undesired consequence of water-fat shift artifacts that are eliminated during data processing. Position, number, and intensity of the ghost spectral lines can be controlled by the suitable choice of spectral bandwidth, number of echo train interleaves, and the number of echoes in each interleave. This study demonstrates that high-spatial resolution EPSI with interleaved flyback readout gradient-echo trains is feasible on standard preclinical scanners.