The nematode genus Acrobeles is composed of two morphological groups distinguished by the presence (‘double’ cuticle) or absence (‘single’ cuticle) of the refringent inner layer of the cuticle. In the present study, four species of this genus, two with ‘single’ cuticle (Acrobeles ciliatus and Acrobeles cylindricus) and two with ‘double’ (Acrobeles aenigmaticus and Acrobeles complexus) are studied from coastal dunes in Spain. This study provides detailed morphological and morphometrical analyses for the four species, while molecular analysis, based on 18S and 28S ribosomal DNA, is provided for A. complexus. The four species are studied with scanning electron microscopy, which is obtained for the first time for A. cylindricus. These analyses revealed morphological and molecular differentiations between both groups, appearing as two related monophyletic entities. The subgenera Acrobeles and Seleborca, formerly considered as separate genera, are proposed to accommodate both groups.