Natural fibre–reinforced composites are more sustainable than other composites with respect to the raw materials. Their properties are attractive due to high specific properties, and especially so wherever high damping is valued. As the interphase between fibre and matrix is the region of highest stresses, a strong bond between fibre and matrix is essential for any composites’ properties. The present study compares two methods of determining the interfacial shear stress in natural fibre–reinforced composites: the single fibre fragmentation test and the single fibre pullout test. The studied composites are flax fibre reinforced epoxy. For a variety of fibre–matrix interaction, the fibres are treated with a laccase enzyme and dopamine, which is known to improve the fibre–matrix shear strength. In the observed samples, single fibre fragmentation test data, i.e. of fracture mode and fragment length, scatter when compared to pullout data. In single fibre pullout tests, the local interfacial shear strength showed a 30% increase in the laccase-treated samples, compared to the control samples. The method also permitted an evaluation of the frictional stress occurring after surface failure.