We assessed the prevalence of Enterobius vermicularis infection and changes in the egg positive rate (EPR) over 1-year time, using the adhesive cellophane-tape perianal swab method in 2,347 preschool children in Muan-gun, Jeollanam-do, Republic of Korea in 2008 and 2009. The overall EPR for E. vermicularis was 4.1% in 2008 and 4.5% in 2009. A study population of 389 children was repeatedly examined for 2 years. Within this group, the EPR in 2009 was twice higher than in 2008, and the EPR of the group of 5-7-year-old children was significantly higher than that of other age groups. Moreover, in the group of 5-7-year-old children in 2009, the rates of positive and negative conversion were significantly higher and lower, respectively, than in other age groups. Conclusively, enterobiasis was prevalent during 2008-2009 among preschool children in Muan-gun, Jeollanam-do, and the increased EPR in 2009 was due to an increase in newly acquired infections among 5-7-year-old children.