Multi-legged locomotion requires appropriate coordination of all legs with coincident ground contact. Whereas behaviourally derived coordination rules can adequately describe many aspects of inter-leg coordination, the neural mechanisms underlying these rules are still not entirely clear. The fact that inter-leg coordination is strongly affected by cut thoracic connectives in tethered walking insects, shows that neural information exchange among legs is important. As yet, recent studies have shown that load transfer among legs can contribute to inter-leg coordination through mechanical coupling alone, i.e., without neural information exchange among legs. Since naturalistic load transfer among legs works only in freely walking animals but not in tethered animals, we tested the hypothesis that connective lesions have less strong effects if mechanical coupling through load transfer among legs is possible. To do so, we recorded protraction/retraction angles of all legs in unrestrained walking stick insects that either had one thoracic connective cut or had undergone a corresponding sham operation. In lesioned animals, either a pro-to-mesothorax or a meso-to-metathorax connective was cut. Overall, our results on temporal coordination were similar to published reports on tethered walking animals, in that the phase relationship of the legs immediately adjacent to the lesion was much less precise, although the effect on mean phase was relatively weak or absent. Lesioned animals could walk at the same speed as the control group, though with a significant sideward bias toward the intact side. Detailed comparison of lesion effects in free-walking and supported animals reveal that the strongest differences concern the spatial coordination among legs. In free walking, lesioned animals, touch-down and lift-off positions shifted significantly in almost all legs, including legs of the intact body side. We conclude that insects with disrupted neural information transfer through one connective adjust to this disruption differently if they experience naturalistic load distribution. While mechanical load transfer cannot compensate for lesion-induced effects on temporal inter-leg coordination, several compensatory changes in spatial coordination occur only if animals carry their own weight.

Many animals establish, learn and optimize routes between locations to commute efficiently. One step in understanding route following is defining measures of similarities between the paths taken by the animals. Paths have commonly been compared by using several descriptors (e.g., the speed, distance traveled, or the amount of meandering) or were visually classified into categories by the experimenters. However, similar quantities obtained from such descriptors do not guarantee similar paths, and qualitative classification by experimenters is prone to observer biases. Here we propose a novel method to classify paths based on their similarity with different distance functions and clustering algorithms based on the trajectories of bumblebees flying through a cluttered environment. We established a method based on two distance functions (Dynamic Time Warping and Fréchet Distance). For all combinations of trajectories, the distance was calculated with each measure. Based on these distance values, we grouped similar trajectories by applying the Monte Carlo Reference-Based Consensus Clustering algorithm. Our procedure provides new options for trajectory analysis based on path similarities in a variety of experimental paradigms.