Necrotizing enterocolitis (NEC ≥ Stage II) is associated with high mortality and morbidity in preterm infants. To assess if introduction of standardized feeding regimen (SFR) and routine probiotic supplementation (RPS) was associated with reduced incidence of NEC in preterm infants in our nursery in a resource limited set up. This was a retrospective cohort study assessing the incidence of NEC ≥ Stage II before (Epoch 1: N = 144) vs. after (Epoch 2, N = 144) implementation of SFR and RPS in preterm infants < 35 weeks. The median (IQR) gestation and birth weight in epoch 1 and epoch 2 was [32 (30, 33.5) vs. 31.5 (30, 34) weeks, p = 0.829], and [1350 (1100, 1700) vs. 1370 (1110, 1550) g, p = 0.363] respectively. Both groups had predominantly outborn infants (Epoch 1: 79.2% vs. Epoch 2: 78.2%; p = 1.00). Multivariate analysis after adjusting for potential confounders found a significantly lower incidence of NEC ≥ Stage II after implementing SFR and RPS (Epoch 1: 17.4% vs. Epoch 2:9.0%, adjusted odds ratio aOR: 0.19; 95% CI: 0.05, 0.71, p = 0.013). The incidence of the composite outcome of 'NEC or Mortality' was also significantly lower after the intervention (Epoch 1: 21.5% vs. Epoch 2: 14.6%; aOR 0.24, 95% CI: 0.07, 0.85, p = 0.027). Introduction of SFR and RPS was associated with significant reduction in NEC ≥ Stage II and the composite outcome of NEC ≥ Stage II /mortality in preterm infants.