The aim of the present study was to investigate whether laterality test results obtained on the ground relate to laterality during riding and to examine the influence of human handedness and horse’s laterality on rein tension. The present study contained an online-survey with 686 riders and 1286 horses as well as the comparison of twelve different methods to investigate horse’s laterality on the ground and during riding between five groups of horses (6.553 horses in total) and rein tension measurements with a group of 88 warmbloods, Quarter Horses and mixed breeds (41 right-lateral, 35 left-lateral and 12 without reported laterality) with 65 riders (49 right-handed, 14 left-handed and 2 ambidextrous) in 110 rides (51 in conventional European riding and 59 in Western riding). Rein tension was analysed using Excel and linear mixed models in SPSS. The relation of different laterality test methods among each other was investigated using cross-tabulations, chi²-tests, phi and Cramer’s V, as well as Pearson-correlations. Heritability was determined using uni- and bivariate linear animal models in DMU6. Laterality test results obtained on the ground did not agree with laterality during riding. Only the rider’s assessment of their horse’s laterality and the lateral displacement of the horse’ hindquarters allowed conclusions on laterality during riding. In most populations the majority of horses had their hindquarters displaced to the right. Based on the rider’s assessment of their horse side preference for dressage tasks, no overall direction of laterality could be documented in any population. Right-lateral and ambidextrous horses were more successful overall. Heritability of the lateral displacement of the hindquarters was high in warmbloods and low to moderate level in Thoroughbreds. In dressage and show jumping there seems to be an advantage for both left-lateral and ambidextrous riders as well as left-lateral horses. A direct relation of the lateral displacement of the hindquarters to sensitive muscle trigger points could not be documented. In addition to the stability of rein tension, two aspects of symmetry (quantitative symmetry between left and right rein tension and temporal symmetry of left and right rein tension peaks) were identified that are influenced mainly by human handedness. Horse’s laterality mainly influenced the magnitude of rein tension as well as the symmetry of the inside versus the outside rein. Right-handed riders depended upon their dominant hand while trying to compensate their horse’s laterality and produced more symmetric rein tension with right-lateral horses. In contrast, left-handed riders reacted to their horse’s non-dominant side, regardless of the horse’s direction of laterality. Less rein tension was applied with a more stable and symmetric contact in Western riding compared to conventional European riding.
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