Although herd size, structure, stability, and social rank among Misaki feral horses have been reported, no studies have been conducted on the affiliative relationships and interactions among members in a Misaki horse herd. The validity of three hypotheses regarding the function of social grooming, the affiliative relationship strengthening hypothesis, the worsened relationship restoring hypothesis, and the grooming parasite removal hypothesis, were tested in a Misaki feral horse (Equus caballus) herd in Cape Toi, Japan. All the nine horses in the “6m” herd were investigated in terms of kinship, grooming, aggression, proximity, social rank, and social network. Mutual grooming occurred only in pairs and was almost perfectly symmetrical. For each member, there was a significant negative correlation between total grooming received from other individuals and self-grooming. Controlling for kinship, there were significant positive partial correlations between mutual grooming and proximity and between aggression and proximity. No correlation was observed between aggression and mutual grooming. The results suggest that mutual grooming symmetry may contribute that both participants simultaneously benefit from parasite removal and strengthen affiliative relationships between seasonally changing herd members; however, mutual grooming did not foster restoring the worsened relationship following aggression promoted by physical proximity. The findings of this study may elucidate the mechanisms by which interactions between herd members are maintained or strengthened.