Objective: To examine if single skin cleansing with 0.25% chlorhexidine affects skin condition, temperature and bacterial colonization in stable preterm (28-36 weeks gestational age) low birth weight (1001-2000 g) infants admitted in a health facility.Methods: Eligible infants were randomized within 3 h of birth into the following three groups: chlorhexidine, normal saline or no skin cleansing. Infants in the first two groups were wiped once with baby wipes containing either 0.25% chlorhexidine or saline. Skin condition, axillary temperature and skin colonization rates in the axilla and the groin were assessed at specified time intervals after intervention.Results: In all, 60 infants were included in the study (20 in each group). Median skin condition scores at 72 and 168 h after the intervention were 2 and 2, respectively, in all three groups. At 30 min after skin cleansing, two infants each in the chlorhexidine and saline cleansing groups and none in the no cleansing group experienced cold stress (36-36.4 1C). There was, however, no difference in mean skin temperature of the groups (36.6 1C). At 24 h, skin colonization rates in the axilla were 22.2, 52.7, and 57.9%, respectively, in the chlorhexidine, saline and no cleansing groups (P ¼ 0.06); skin cleansing with chlorhexidine reduced the incidence of colonization by 62% compared with no cleansing (relative risk (RR): 0.38, 95% confidence interval (CI): 0.15, 0.98), but there was no significant reduction when compared with saline cleansing (RR: 0.42; 0.16-1.10). Axillary colonization rates at 72 h and colonization at the groin at 24 and 72 h were not significantly different across the three groups.Conclusion: Single skin cleansing with 0.25% chlorhexidine did not adversely affect skin condition or temperature in hospitalized preterm infants and reduced axillary-skin colonization at 24 h after the intervention. Trials are needed to evaluate the efficacy of such an intervention on the incidence of infections in preterm neonates.