Relatively low sperm count was reported among young Spanish men in 2013. Several potential culprits have been suggested as explanations for reported trends in sperm counts in Western men, including lifestyles. Although controversial, some studies suggest that semen parameters, such as low sperm motility or abnormal morphology, may be associated with low serum vitamin D levels. To evaluate associations between semen parameters and reproductive hormones and serum 25-hydroxyvitamin D (25OHD) status in young Spanish men and to examine these associations in relation to dietary intake of vitamin D. This cross-sectional study includes 198 university students recruited in 2010-2011 in southern Spain, who provided samples of blood and semen and food frequencies. Semen quality was evaluated by measuring volume, concentration, sperm counts, motility, and morphology, according to the WHO guidelines. Serum samples were analyzed for total 25OHD and reproductive hormones, including FSH, LH, testosterone, inhibin B, and estradiol. Dietary vitamin D intake was assessed using a validated food frequency questionnaire. Associations with semen quality and reproductive hormones were examined using linear regression, adjusting for potential confounders. Almost all men had adequate levels of serum vitamin D - only three men (1.5%) were vitamin D deficient (<30nmol/L) and 17% were insufficient (<50nmol/L). However, dietary vitamin D intakes were relatively low (below recommended 600 IU/day in 99% of men). Neither dietary intake nor serum vitamin D levels were associated with any sperm parameter or any reproductive hormone (all p≥0.09). We did not observe an association between vitamin D status and any reproductive parameter in our study population. Our results suggest that serum vitamin D levels are sustained in Spanish men despite low dietary intake and therefore low vitamin D does not explain the poor semen quality previously observed in these young Spanish men.