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From January to May 2012, a cluster of 12 cases of necrotizing enterocolitis (NEC) occurred in the Neonatal Intensive Care Unit of one of the Hospitals in Salerno, Italy. Six of twelve infants developed severe NEC (stage II – III). All cases, but one, were preterm of less than 37 week gestation (91.7%); five cases (41.7%) had a very low birth weight (VLBW) less than 1,500 g and five between 1500 g and 2500 g (Low birth weight - LBW). The main risk factors associated with NEC was central venous line. Klebsiella pneumoniae was isolated from the clinical samples of 10 cases, of which 9 (90%) were extended-spectrum β-lactamase (ESβL) producers. K. pneumoniae were also isolated from various sites of environmental samples, suggesting the causal role of this pathogen in the development of NEC in addition to other risk factors.We describe an outbreak of NEC in a tertiary care neonatal unit, along with the case-control study performed and microbiological investigation to identify noninfectious and infectious risk factors potentially involved in the outbreak.