Experimental studies have shown that anogenital distance (AGD) at birth reflects androgen concentrations during prenatal development and predicts adult AGD. In male rodents, shortened AGD is associated with compromised reproductive function. The aim of this study was to examine associations between AGD measures and semen quality in male partners of subfertile couples. All men were attending an infertility service and provided a semen sample and completed epidemiological questionnaires on lifestyle and general health. Two variants of AGD [from the anus to the posterior base of the scrotum (AGDAS) and to the cephalad insertion of the penis (AGDAP)] were assessed in 91 men. Semen parameters (semen volume, sperm concentration, total sperm counts, motility, and morphology) were determined following WHO guidelines. Associations between AGD measures and semen quality were tested using multiple regression analyses controlling for appropriate covariates. Significant positive associations between AGDAS measures and sperm concentration, total sperm count, and total sperm motile count were detected (p-values<0.05). This study represents the first analysis of AGD in potential infertile European men showing an association between perineal length and semen parameters. However, it is still uncertain the utility to clinical practice, therefore further studies are warranted to confirm and extent these findings.