The present study investigated the relationship between hyperglycemia and both serum carotenoids and intake of vegetables and fruits. Subjects with a history of diabetes mellitus (DM group, n = 133) or with hyperglycemia diagnosed using a 5.6% cutoff value for hemoglobin A1c (High HbA1c group, n = 151) were recruited from among inhabitants of a rural area in Hokkaido, Japan. Intake frequencies of vegetables and fruits were assessed using a questionnaire administered by public health nurses. Serum levels of carotenoids and retinol were measured using high-performance liquid chromatography. The relationships between high HbA1c or DM and both serum carotenoids and intake frequencies of vegetables and fruits were analyzed using logistic regression modeling for a case-control study; each case (High HbA1c or DM) was matched to two controls (healthy subjects without any history of disease) matched for sex and age (within 3 years). The odds ratio (OR) for high HbA1c was 0.49 (95% confidence interval: 0.29-0.85) on high intake frequency of carrot and pumpkin and the OR for DM was 1.21 (95% Cl: 0.79-1.84). No significant relationships were observed between high HbA1c and intake frequencies of other vegetables and fruits. The ORs on high serum levels of alpha- and beta-carotenes, lycopene, beta-cryptoxanthin and zeaxanthin and lutein were 0.38 (0.22-0.65), 0.35 (0.21-0.59), 0.57 (0.35-0.93), 0.35 (0.20-0.59), and 0.88 (0.54-1.46) for high HbA1c, respectively. In conclusion, intake of vegetables and fruits rich in carotenoids might be a protective factor against hyperglycemia.