BACKGROUND: The increase in invasive fungal infections (IFIs) in neonatal intensive care unit (NICU) is jeopardizing the survival of preterm neonates. Probiotics modulating the intestinal microflora of preterm neonates may minimize enteral fungal colonization. AIMS: This study was to examine whether probiotic supplementation in neonates reduced fungal septicemia. MATERIALS AND METHODS: This prospective, randomized, double blind trial investigating the supplementation of preterm infants with a probiotic was done from May 2012 to April 2013, with 112 subjects randomized into two groups. PRIMARY OUTCOME: Decreased fungal colonization in gastrointestinal tract. Others: Incidence of late onset septicemia; duration of the primary hospital admission; number of days until full enteral feeds established. RESULTS: Full feed establishment was earlier in probiotics group compared to placebo group (P = 0.016). The duration of hospitalization was less in the probiotic group (P = 0.002). Stool fungal colonization, an important outcome parameter was 3.03 ± 2.33 × 10(5) colony formation units (CFU) in the probiotics group compared to 3 ± 1.5 × 10(5) CFU in the placebo group (P = 0.03). Fungal infection is less in the study group (P = 0.001). CONCLUSION: The key features of our study were reduced enteral fungal colonization, reduce invasive fungal sepsis, earlier establishment of full enteral feeds, and reduced duration of hospital stay in the probiotics group.