Objective To explore the feasibility of two-dimensional speckle tracking imaging evaluating healthy adult canines’ left ventricular fractional strain and rotation,and the relationship between strain parameters and left ventricular systolic function.Methods We analyzed the fractional strain and rotation(RS,CS,Rot)of 27 healthy adult canines by using 2DT online analysis software,and evaluated the correlation of these parameters and left ventricular systolic function with Simpson method.Results Left ventricular systolic RS was positive，and RS of papillary muscle level were significantly increased compared with basal and apical level.CS was negative,and the absolute value of the basal segment to apical segment gradually increased.The maximum Rot level was apical level,while the minimum level was papillary muscle level (P<0.05).RS,CS and Rot of different segments of inner layers showed higher than those of the outer layers (all P＜0.05).GRS,GCS and Rot correlated well with left ventricular systolic function by Simpson method (r values were 0.72,0.87,0.80 respectively,all P <0.05).Conclusions Two-dimensional speckle tracking imaging can evaluate healthy adult canine’s left ventricular fractional strain and rotation accurately.Fractional global strain and rotation reflect left ventricular systolic function well.